KR100987302B1 - Cosmetic composition having anti-wrinkle effect containing the horse placental extract - Google Patents

Abstract

PURPOSE: A cosmetic composition containing natural extract which promotes collagen synthesis is provided to relieve wrinkle and to ensure skin elasticity. CONSTITUTION: A composition for anti-wrinkle contains 0.0005-10 weight% of horse placenta extract as an active ingredient. A method for manufacturing horse placenta extract comprises: a step of adding 0.2-0.6 weight parts of papain, protease, peptidase, pepsin or mixture thereof and 65-100 weight parts of water to 30-40 weight parts of horse placenta at 60-70°C; a step of adding 0.05-0.2 weight parts of protease to the reactant and performing proteolytic reaction at 60-70°C; and a step of removing smell and color under vacuum pressure. The composition for anti-wrinkle is used in the form of cream, essence, ointment, spray, cleansing foam, cleansing water, pack, body oil, lotion, foundation, lip stick, mascara, makeup base, and sun screen.

Description

Cosmetic composition having anti-wrinkle effect containing the horse placenta extract

The present invention relates to a wrinkle improvement cosmetic composition containing placenta extract. More specifically, the present invention relates to a skin wrinkle improvement cosmetic composition containing a placental extract having excellent skin aging and anti-wrinkle effect, and at the same time, excellent in vivo safety.

Skin aging is classified into intrinsic aging, a natural aging that comes naturally with age, and photo aging observed in the face, neck, and hands, which are parts of the body exposed to light (ultraviolet, UV) for a long time. do.

Most of the endogenous aging phenomenon is observed in the skin of people who are not exposed to sunlight, and the clinical characteristics are mostly mild forms of fine wrinkles, dry skin, and reduced elasticity. This is because the action of fibroblasts, which form the basis of skin, declines, the number of cells decreases, and the amount of extracellular matrix protein fibers such as collagen, elastin, and fibrillin decreases. Due to this structure, the skin tissue is loosened, the elasticity is reduced, the loss of moisture in the skin cells is increased and the structure of the stratum corneum changes. Limiting the function of fibroblasts limits collagen production and impairs the action of elastin to determine skin elasticity. Protein biosynthetic activity of fibroblasts is known to decrease with age (PCT WO 92/20341). The skin covers the surface of the body and is divided into epidermis and dermis. The epidermis serves to protect internal organs, and skin cell proliferation is caused by the activity of the fibroblasts in the dermis. In addition, the dermis is composed of collagen and elastin that are woven into a net shape (Braverman, J. Invest. Dermatol., 78, 434-443, 1982) to maintain the elasticity of the entire skin. .

Photoaging is repeatedly exposed to light, not only fine wrinkles but also thick and deep wrinkles occur, and pigment diseases such as solar lentigo, in which irregular pigmentation occurs, occur together. In addition, the skin becomes very rough and dry, the elasticity is reduced, the skin is drooping phenomenon occurs.

A typical symptom of skin aging is wrinkles. There are various theories about the origin of wrinkles, but it is not known exactly. Until now, the theory that the main causes of damage to the matrix proteins such as collagen and elastic fibers in the skin is accepted. In addition, the distribution of facial muscles, muscle movements, genetic factors, ultraviolet rays, smoking, drinking, drugs, food, sleep, heat It is a common opinion of dermatologists that a combination of factors, including oxidative damage and hormonal changes (menopause), is difficult to explain in any one theory.

Cosmetics to improve skin wrinkles have been developed for a long time. Especially, in the case of skin wrinkles generated in women, it is very sensitive and UV blockers or moisturizers for the occurrence of skin wrinkles that are thought to be caused by external factors. The back is pouring with various products.

One of the targets to improve skin wrinkles is collagen. Collagen (collagen) is a major protein that forms the skin with elastin, the amount of collagen decreases with age and UV exposure. This is because the collagen protein decreases and the skin loses its elasticity and wrinkles occur.

Retinoic acid, retinol and vitamin C are examples of skin wrinkle improving substances that promote collagen synthesis. However, each of the substances has a disadvantage, Retinoic acid is a strong irritant to the skin, Retinol is not easy to oxidize or cause skin problems because of its low stability. In addition, the case of vitamin C is also easily deteriorated by air, temperature, etc., is known to have a lot of stability problems.

Collagen synthesis has been studied a lot in Korea, and is applied to a variety of cosmetics are sold. As an example described for the anti-wrinkle cosmetic composition that promotes collagen synthesis, Landing (Radix Phytolacca esculenta) or Semen Cuscutae extract (Korean Patent Publication No. 2000-13588), Agastache rugosa and Aroma (Aquilaria agallocha) Roxb.) Extract (Korean Patent Publication No. 2000-18467), Ecklonia cava extract (Korean Patent Publication No. 2002-85997), Malletus japonicus extract (Korean Patent Publication No. 2005-39327), etc. In addition, the cosmetics using natural extracts are less toxic and have the advantage of being able to be produced in large quantities, and thus have many advantages as cosmetics.

Horse placenta contains essential amino acids, nucleic acids such as RNA and DNA, superoxide dismutase (SOD), hyaluronic acid, antioxidants, immune cofactors (cytocine), placental peptide and insulin-like growth. It contains growth factors and cytokines such as insulin-like growth factor, epidermal growth factors (EGFS) and unique senescent cell activating factors (SCAFS). It is known to be useful for augmentation and the like. In addition, the inventors confirmed that the extract extracted from the placenta is effective in inhibiting and improving skin wrinkles.

In particular, the placenta extract has an inherent off-flavor, so it is insufficient to use it as a food or cosmetics. In the manufacturing method of Korean Patent No. 932183, a method of adsorbing off-flavor of the placenta extract using activated carbon under vacuum pressure conditions It was used as a food using. In the present invention, the placenta extract prepared by using the manufacturing method of the Korean Patent No. 932183 and the improved method thereof is used for cosmetics. Instead of performing the adsorption process under vacuum pressure by the improved method, it is possible to prepare a better placenta extract for use as a cosmetic by removing odor using activated carbon under high-purity nitrogen conditions.

An object of the present invention to provide a cosmetic composition for wrinkle improvement and wrinkle relief containing horse placenta (Horse placenta) extract.

Another object of the present invention to provide a composition for promoting collagen synthesis, characterized in that it contains a placenta extract.

Wrinkle improvement composition of the present invention is characterized in that it contains horse placenta (Horse placenta) extract as an active ingredient.

The composition containing the placenta extract can be used with a carrier usable in the cosmetic composition.

"Wrinkle improvement" of the present invention means reducing the depth of wrinkles, the number and area of wrinkles, "wrinkle improvement", "wrinkle relief" and "skin elasticity" are used synonymously.

The horse placenta extract of the present invention uses the preparation method of Korean Patent No. 932183 in thawing and washing, preparation of hydrolyzate using enzyme, filtration, separation and purification, membrane filtration, etc., but uses some modified conditions.

To this end, the proteolytic reaction is performed by adding protease and water to the horse's placenta, and further proteolytic reaction is added to the reaction product, followed by secondary proteolytic reaction, and then activated carbon is added at 0.1 to 0.5 atm. It characterized in that it comprises the step of removing the smell and color under vacuum pressure or high purity nitrogen (99.9%). The optimum vacuum pressure of the refining process is preferably 0.1 atm to 0.5 atm, more preferably 0.2 to 0.3 atm.

Preferably, the proteolytic reaction is performed by adding 0.2 to 0.6 parts by weight of protease and 65 to 100 parts by weight of water to 30 to 40 parts by weight of placenta, and 0.05 to 0.2 parts by weight of protease for perfect proteolysis. After the addition of additional parts to the second proteolytic reaction, 1.0 to 1.5 parts by weight of activated carbon is added to remove the smell and color.

In order to obtain the horse placenta extract according to the present invention, the normal placenta of horses, which have been specifically tested by a veterinarian, are put in a freezer or the like and handwritten. The frozen horse placenta is thawed and pulverized to facilitate proteolysis, and the ground horse placenta is washed until blood is removed. At this time, if the blood is not removed, the odor component generated by the blood component is a washing operation is essential.

In this way, the cleanly washed horse placenta is dehydrated using a filter, and proteolytic reaction is performed by adding protease and water. The proteolytic reaction at this time is carried out in two steps. The reason for this is that the proteolytic reaction does not proceed smoothly in the present invention, and the process takes a long time, and in order to improve the disadvantage of using an excessive protease, the decomposition reaction is divided into two times even if a small amount of enzyme is used. Giving a new activated enzyme surface gives a significant reduction in process time, resulting in increased productivity and stability. As the protease, papain, protease, peptidase, pepsin, or a mixture thereof may be used.

The initial placental content of the total reactant is preferably included to be 30 to 40 parts by weight, preferably 33 to 38 parts by weight, more preferably 33 to 35 parts by weight. If the placenta content is included in less than 30 parts by weight, there is a problem that the content of the useful component is low, when it is included in excess of 40 parts by weight it is uneconomical because the hydrolysis efficiency does not increase significantly compared to the excess amount.

The proteolytic reaction time is preferably reacted for a total of 5 to 12 hours, more preferably 7 to 9 hours. In the case of reacting for less than 7 hours, the hydrolysis reaction does not occur sufficiently, and in the case of reacting for more than 9 hours, it is uneconomical for more than the time required for the reaction.

The number of proteolytic reactions is preferably 1 to 2 times, and it is uneconomical because 3 times does not increase the efficiency and consumes only enzymes.

50-80 degreeC is preferable and, as for the temperature of the said proteolysis reaction, 60-70 degreeC is more preferable. If the reaction is less than 60 ℃ there is a problem that the reaction does not occur properly, if the reaction is exceeded 70 ℃ due to the problem of inactivation of the enzyme is difficult to completely occur hydrolysis reaction has the disadvantage that the useful components are denatured. In addition, after this process, the complete protein hydrolysis can be confirmed by the biuret reaction and the acetic acid reaction with tricholo.

The water content of the activated carbon used in the purification process for high purity purification is preferably 1% to 5%, more preferably 3% or less.

In addition, vacuum or high-purity nitrogen conditions are very important for high-purity purification using activated carbon, which is a cause of odor or odor caused by activated charcoal under vacuum or high-purity nitrogen conditions because purification with activated carbon cannot remove the perfect odor under normal conditions. It is good to adsorb and remove materials. In particular, the temperature is preferably treated under agitation at a high temperature of 60 to 80 ℃, which is not only increases the adsorption rate of activated carbon but also volatile substances that cause odors when treated under high pressure and vacuum pressure or high purity nitrogen conditions. This can be effectively removed.

In the present invention, after the purification process may further include a filtration step for removing the solid matter and activated carbon. For this purpose, it is preferable to perform filtration one or more times using a 0.1 to 3 μm filter. If the pore size of the filter is less than 0.1μm, the filter is very precise and extremely expensive (ultrafiltration method), so there is a problem of low economic efficiency. There is a problem that this is high.

The filtration step may be performed about 1 to 3 times, in the embodiment of the present invention, the first filtration is performed by 2 μm filter, the second filtration by 0.45 μm filter, and the third filtration is performed three times by using 0.2 μm filter. do.

In order to produce the finished product of the composition obtained by the above method, in order to fill the container of the composition of the present invention, after washing the container with ethanol and UV sterilization drying and filling the sterilized container with the composition of the present invention, 121 Packaged by autoclaving at 30 ° C. for 30 minutes.

Horse placenta extract of the present invention may be prepared in a powder state by an additional process such as distillation under reduced pressure and lyophilization or spray drying.

The cosmetic composition containing the placenta extract of the present invention is prepared according to a conventional method of cosmetic preparation.

Horse placenta extract is contained in an amount of 0.0001 to 20% by weight, preferably 0.0005 to 10% by weight based on the total weight of the cosmetic composition. The cosmetic composition containing the placenta extract of the present invention for use on skin or hair is formulated with a pharmaceutically or dermatologically acceptable excipient and a lotion, cream, essence, ointment, spray, cleansing foam, cleansing water, pack, body It can be applied to oils, lotions, foundations, lipsticks, mascaras, makeup bases, sunscreens, makeup removers and cleaners and the like. Excipients can include pharmaceutically acceptable emollients, skin penetration enhancers, colorants, fragrances, emulsifiers, thickeners, solvents.

The cosmetic composition that can be prepared using the placenta extract of the present invention will be described by taking a lotion, cream, solution, gel or solid as an example.

Lotion, which has an intermediate character of lotion and cream, is more fluid and less oily than the cream. The main components are emulsifiers, emollients, antioxidants, thickening agents, pH buffers and solvents, etc., the emulsifier serves to mix the oil phase and the water phase of the cosmetic composition. Emulsifiers can include both nonionic, anionic and cationic. Emollients may include conventional emollients such as fatty acid esters, fatty alcohols, petroleum based, propylene glycol, glycerin, mercury, waxes, fatty acids, fatty alcohol ethers, acetoglycerides, lanolin and derivatives thereof. The thickening agent may include crosslinked carboxypolymethylene polymer, cellulose, polyethylene glycol, gum. Antioxidants used to improve stability include ascorbic acid and derivatives thereof, and may also include tocopherol and derivatives thereof. pH buffers are used to adjust the pH of cosmetic compositions, including acetates, phosphates, citrate, carbonates and the like. As the solvent, purified water, alcohol, a mixture of purified water and alcohol, hot water and the like can be used.

 The lotion of the present invention may contain 0.001-20% by weight of placenta extract. The emollient may contain 0.01 to 30% by weight, preferably 0.10 to 20% by weight. The emulsifier preferably contains 0.01 to 10% by weight, preferably 0.1 to 5% by weight. Preservatives are preferably added at 0.01 to 4.0% by weight, flavoring at 0.02 to 2.0% by weight, and purified water at 65 to 90% by weight.

Cream of the present invention may contain a placenta extract 0.001 ~ 20% by weight. The emollient may contain 0.01 to 50% by weight, preferably 0.10 to 40% by weight. The emulsifier is preferably 0.01 to 20% by weight, preferably 0.1 to 10% by weight. Preservative is preferably 0.01 to 4.0% by weight, fragrance is preferably added at 0.02 to 2.0% by weight, purified water is preferably added to 50 to 90% by weight. Cream preparation can increase the oil and moisturizer content of the emollient compared to lotions.

Solutions may include softening water, astringent water, essences and packs.

The flexible longevity composition of the present invention may contain the placenta extract in an amount of 0.001 to 10% by weight, preferably 0.01 to 50% by weight. Skin softener is preferably 0.1 to 10.0% by weight, preferably 0.05 to 5.0% by weight is appropriate. As a preservative, 0.01 to 4.0% by weight, fragrance is preferably added at 0.02 to 2.0% by weight, and 70 to 90% by weight of purified water may be added as a solvent.

The convergent longevity composition may contain the placenta extract in an amount of 0.001 to 10% by weight, preferably 0.05 to 50% by weight. Preservative is preferably 0.01 to 4.0% by weight, fragrance is added to 0.02 to 2.0% by weight, 70 to 90% by weight of purified water is suitable as a solvent, 1.0 to 20% by weight of ethanol can be added. have.

Essence contains a high concentration of cosmetic ingredients, has the effect of moisturizing, skin protection and nutrient supply, the main components may include moisturizers, skin softeners, solvents, emulsifiers and thickeners.

The pack facilitates the absorption of nutrients, the main ingredients may be a solvent such as a film forming agent, a moisturizer, an emulsifier and ethanol.

Gel compositions can be prepared by mixing thickeners in the compositions of the lotions, creams, essences and packs described above.

Cosmetic composition containing the placenta extract of the present invention can be applied to the body parts, especially the face, neck, hands, feet, arms, legs, etc. to reduce wrinkles.

The cosmetic composition of the present invention may be used daily and may also be used for an indefinite period of time. Preferably, the amount of use, the number of times and the period of use can be adjusted according to the age, skin condition or skin type of the user, the concentration of the placenta extract.

The treatment method of the composition of the present invention can be treated by applying to the surface of the human skin in the form of a cosmetic composition such as cream, lotion, lotion, essence and pack.

Horse placenta extract of the present invention is prepared using a method for producing horse placenta extract of Korean Patent No. 932183 or an improved method thereof. In Korean Patent No. 932183, the high-odor adsorption of horse placenta extract is performed under vacuum pressure. In the present invention, a method using vacuum pressure or high purity nitrogen (99.9%) was performed. This is because a component that causes high odor of the horse's placenta has a high volatility characteristic and thus can quickly remove the volatile substance under vacuum pressure or high purity nitrogen condition. In particular, high-purity nitrogen conditions are better for use as cosmetics because more odor of placenta extract is removed than vacuum conditions. In addition, the present invention reduces the number of reactions of protease, increases the concentration of enzyme, and reduces the production time and lowers the production cost than the preparation method of Korean Patent No. 932183.

The present invention provides a composition for promoting collagen synthesis and wrinkle improvement cosmetic composition comprising the placenta extract as an active ingredient. The composition of the present invention exhibits an excellent anti-wrinkle effect by inducing collagen synthesis promotion through a molecular level mechanism. This will not only replace the wrinkle improvement raw materials currently used in cosmetics, but also enhance international competitiveness and enhance the efficacy and added value of placenta.

1 is a graph showing the collagen synthesis enhancing effect of the horse placenta extract, pig placenta extract and retinol of the present invention in fibroblast WI-38 (Human Fibroblast Cell Line).

<Example 1. Preparation of horse placenta extract prepared by enzyme treatment and ultrapure nitrogen adsorption process-I>

To prepare the horse placenta extract, first thawed the frozen horse placenta 1.5kg and then crushed as finely as possible through a cutter. As the cutter, a grinder which is commonly used for scissors or cooking may be used. The ground placenta was put in a stirrer and washed until the blood was removed, followed by dehydration. Thereafter, 9 g of papain and 3 kg of distilled water were added to 1.2 kg of the dehydrated placenta and reacted at 60 ° C. for 5 hours so that the placenta was completely dissolved in water. For complete digestion 3 g of papain was added to the digested solution and stirred at 70 ° C. for 2 hours. After stirring, complete protein hydrolysis was confirmed by burette reaction and trichloroacetic acid reaction. 30 g of activated carbon was added for color and odor suitable for cosmetic raw materials and stirred for 30 minutes under reaction temperature of 70 ° C. and high purity nitrogen (99.9%). The color and smell were removed. The activated carbon was dried for 12 hours in a vacuum oven at 120 ℃ for 12 hours after taking a small amount of samples to measure the moisture measurement was 3% or less was used as a raw material for the purification process. The activated carbon purified solution was filtered to remove activated carbon. The primary filtration was performed using a 2 μm filter, the secondary filtration using a 0.45 μm filter, and the tertiary filtration using a 0.2 μm filter.

<Example 2. Preparation of Horse Placenta Extract Prepared by Enzyme Treatment and Ultrapure Nitrogen Adsorption Process-II>

A horse placenta extract was prepared in the same manner as in Example 1, but a protease was used instead of papain.

<Example 3. Preparation of horse placenta extract prepared under 0.2 atm pressure rather than ultrapure nitrogen during the adsorption process>

Horse placenta extract was prepared in the same manner as in Example 1, but the placenta extract was prepared by performing an adsorption process using activated carbon under 0.2 atm of non-pure nitrogen.

Example 4 Cream Containing Horse Placenta Extract Prepared by Enzyme Treatment and Ultrapure Nitrogen Adsorption Process-100g in total>

Placenta extract of Example 1 ........................ 2.0 g

Paraben ......................... 0.2 g

Allantoin ......................................... 2.0 g

Betaine ......................................... 1.0 g

Sodium hyaluronate ......................................... 0.13 g

Tocopherol Acetate ......................................... 10 g

Shea butter 10 g

Jojoba oil ......................................... 1.7 g

Preservative ............... 0.4 g

Fragrance ... ... 0.2 g

Distilled Water ... g

<Example 5. Gel containing horse placenta extract prepared by enzymatic treatment and ultrapure nitrogen adsorption-total 100g>

Placenta extract of Example 1 ........................ 2.0 g

Ammonium Acrylodimethyldimethyl Taurate ..................... 2.0 g

Glycerin ......................................... 2.0 g

Butylene Glycol ......................................... 3.0 g

Propylene Glycol ......................... 2.0 g

Sodium Hyaluronate ......................................... 5.0 g

Glyceris-26 ......................................... 2.0 g

Ethanol ......................................... 1.5 g

Methylparaphene ......................... 0.2 g

PPG-26-Buteth-26 ... 1.0 g

PEG-40 Hydrogenate Castor Oil ......... 0.5 g

Phenyloxyethanol ......................................... 0.5 g

Phenyltrimethicone ................................. 0.7 g

Bis-PEG-18 methyl ether dimethyl saline ......... 0.3 g

Preservative ............... 0.4 g

Fragrance ... ... 0.2 g

Distilled Water ... g

<Example 6. Essence containing horse placenta extract prepared by enzymatic treatment and ultrapure nitrogen adsorption-total 100g>

Placenta extract of Example 1 ........................ 2.0 g

Glycerin ..................... 10.0 g

Betaine ......................................... 5.0 g

PEG 1500 ....................................... 2.0 g

Allantoin ......................................... 0.1 g

DL-panthenol ...................................... 0.3 g

EDTA-2Na ........................................ 0.02 g

Benzophenone-9 ..................................... 0.04 g

Hydroxyethyl cellulose ................................... 0.1 g

Sodium Hyaluronate ......................... 8.0 g

Carboxy vinyl polymer ..................... 0.2 g

Triethanolamine ..................... 0.18 g

Octyldodecanol ......................................... 0.3g

Octyldodeceth-16 ....................................... 0.4g

Ethanol ... g

Preservative ............... 0.4 g

Fragrance ... ... 0.2 g

Distilled Water ... g

<Example 7. Pack containing horse placenta extract prepared by enzymatic treatment and ultrapure nitrogen adsorption process-total 100g>

Placenta extract of Example 1 ........................ 2.0 g

Polyvinyl alcohol ......................................... 15.0 g

Cellulose Gum ........................ 0.15 g

Glycerin ......................................... 3.0 g

PEG-1500 ......................................... 2.0 g

Cyclodextrin ........................... 0.15 g

DL-panthenol ......................................... 0.4 g

Allantoin ......................................... 0.1 g

Monoammonium glycyrrhinate ...................... 0.3 g

Nicotinamide ......................................... 0.5 g

Ethanol ... g

Preservative ............... 0.4 g

Fragrance ... ... 0.2 g

Distilled Water ... g

Example 8 Preparation of Cream Containing Horse Placenta Extract Prepared at 0.2 Atmospheric Pressure Conditions Instead of Ultrapure Nitrogen During Adsorption>

Preparation of the cream in the same manner as in Example 4 but instead of placenta extract of Example 1 was used the placenta extract of Example 3.

Comparative Example 1. Preparation of horse placenta extract prepared using an acid catalyst

In the same manner as in Example 1 to prepare a placenta extract, but instead of papain was added 12 g of concentrated hydrochloric acid reacted at 100 ℃ to prepare a placenta extract.

Comparative Example 2. Preparation of Horse Placenta Extract Prepared Using Alkali Catalyst

To prepare horse placenta extract in the same manner as in Example 1, but instead of papain was added to KOH 12g to react at 100 ℃ to prepare a horse placenta extract.

Comparative Example 3. Preparation of horse placenta extract prepared by performing only the first proteolysis reaction

The horse placenta extract was prepared in the same manner as in Example 1, but the horse placenta extract was prepared by performing only the first proteolysis without performing the second proteolysis.

<Comparative Example 4. Preparation of horse placenta extract prepared by using organic solvent precipitation instead of enzyme treatment>

Prepare horse placenta extract in the same manner as in Example 1, but put the same amount of acetone in place of papain and distilled water and reacted at room temperature to dry acetone at 80 ℃ in a vacuum dryer to prepare a horse placenta extract.

Comparative Example 5. Preparation of Placenta Extract Prepared by Ultrasonic Wave instead of Enzyme Treatment>

Preparation of placenta extract in the same manner as in Example 1, but instead of treating the enzyme was treated with ultrasonic waves 10 times at intervals of 30 seconds to prepare a placenta extract.

Comparative Example 6. Preparation of Horse Placenta Extract Prepared by Heat Treatment Instead of Enzyme Treatment>

In the same manner as in Example 1 to prepare a placenta extract, but instead of treating the enzyme was heated to 50 minutes at 50 ℃ to prepare a placenta extract.

Comparative Example 7. Preparation of Horse Placenta Extract Prepared Without Adsorption Process>

A horse placenta extract was prepared in the same manner as in Example 1, but the adsorption process using activated carbon was not performed under ultrapure nitrogen.

Comparative Example 8. Preparation of Porcine Placenta Extract Using Porcine Placenta

 Placenta extract was prepared in the same manner as in Example 1, but pig placenta extract was prepared using pig placenta instead of horse placenta.

Comparative Example 9. Preparation of Cream Containing Horse Placenta Extract Prepared by Acid Catalyst

To prepare a cream in the same manner as in Example 4 but using a placenta extract of Comparative Example 1.

Comparative Example 10 Preparation of Cream Containing Horse Placenta Extract Prepared Using Alkali Catalyst

Cream was prepared in the same manner as in Example 4, but was used for placenta extract of Comparative Example 2.

<Comparative Example 11. Preparation of cream containing horse placenta extract prepared by performing only the first proteolytic reaction>

To prepare a cream in the same manner as in Example 4 but was used for placenta extract of Comparative Example 3.

Comparative Example 12 Preparation of Cream Containing Placenta Extract Prepared Using Organic Solvent Precipitation Instead of Enzyme Treatment

Cream was prepared in the same manner as in Example 4, but was used for placenta extract of Comparative Example 4.

Comparative Example 13. Preparation of Cream Containing Placental Placenta Extract Prepared by Ultrasound Instead of Enzyme Treatment>

Cream was prepared in the same manner as in Example 4, but was used for placenta extract of Comparative Example 5.

Comparative Example 14. Preparation of Cream Containing Placental Placenta Extract Prepared by Heat Treatment Instead of Enzyme Treatment>

To prepare a cream in the same manner as in Example 4 but was used for placenta extract of Comparative Example 6.

Comparative Example 15 Preparation of Cream Containing Placenta Extract Prepared Without Adsorption Process

To prepare a cream in the same manner as in Example 4 but using a placenta extract of Comparative Example 7.

Comparative Example 16 Preparation of Cream Containing Porcine Placenta Extract

To prepare a cream in the same manner as in Example 4, but was used for placenta extract of Comparative Example 8.

Comparative Example 17 Preparation of General Cream

Cream was prepared in the same manner as in Example 4, but the placenta extract was not used.

Experimental Example 1: Enhance Collagen Synthesis and Confirm Cytotoxic Effects

In order to confirm collagen synthesis enhancement and cytotoxic effects, the human fibroblast cell line WI-38 (Human Fibroblast Cell Line) was inoculated with human MEM (Minimum Essential Media) medium and incubated at 37 ° C for 24 hours. The horse's placenta extract in each condition was diluted twice with MEM solution without FBS, and the dilutions were diluted stepwise by 10 times so that the concentration range was 5 × 10 ⁻¹ to 5 × 10 ⁻⁵ % (w / v). Subsequently, after treating the placental extract of each condition to the cultured cells, the amount of collagen produced using the PIP ELISA Kit (TAKTRA, Cat. MK101) was measured and the results are shown in Table 1 and FIG. 1.

 The results of Table 1 showed that the production efficiency of collagen by concentration for the placenta extract of the present invention was increased by up to 52.3% more than the control group that did not process the placenta extract at all. It showed more than twice the efficacy of 24% collagen synthesis ability of the retinol which is currently widely used, and was significantly higher than the placenta extract in Comparative Examples 1 to 8 conditions. As a result, it can be seen that the horse's placenta extract of the present invention has an excellent collagen synthesis promoting effect. In addition, even if the concentration of placenta extract increased, fibroblast WI-38 did not show any cell killing effect (checked cell number using trypan blue).

Condition

Collagen Synthesis Ratio (%) 5 × 10 ^-5 w / v%
extract 5 × 10 ^-4 w / v%
extract 5 × 10 ^-3 w / v%
extract 5 × 10 ^-2 w / v%
extract 5 × 10 ^-1 w / v%
extract Example 1
Placenta Extract
2.4 ± 0.9
17.2 ± 0.5
35.2 ± 0.0
43.2 ± 0.1
52.3 ± 0.2 Example 2
Placenta Extract
2.7 ± 0.6
19.2 ± 0.8
33.2 ± 0.5
41.7 ± 0.0
50.2 ± 0.5 Example 3
Placenta Extract
1.9 ± 0.6
16.7 ± 0.3
28.7 ± 0.9
37.2 ± 0.4
45.2 ± 0.2 Of Comparative Example 1
Placenta Extract
2.8 ± 0.7
5.7 ± 0.5
8.4 ± 0.4
17.7 ± 0.4
35.2 ± 0.3 Of Comparative Example 2
Placenta Extract
2.7 ± 0.6
4.6 ± 0.4
7.3 ± 0.5
16.4 ± 0.3
34.3 ± 0.4 Of Comparative Example 3
Placenta Extract
1.6 ± 0.5
4.1 ± 0.3
6.7 ± 0.3
15.6 ± 0.5
28.4 ± 0.5 Of Comparative Example 4
Placenta Extract
1.8 ± 0.6
5.2 ± 0.4
7.4 ± 0.7
14.4 ± 0.4
29.5 ± 0.6 Of Comparative Example 5
Placenta Extract
2.5 ± 0.7
5.3 ± 0.5
8.2 ± 0.5
18.2 ± 0.6
33.7 ± 0.2 Of Comparative Example 6
Placenta Extract
2.5 ± 0.5
6.5 ± 0.5
8.3 ± 0.6
17.5 ± 0.5
32.3 ± 0.3 Of Comparative Example 7
Placenta Extract
2.6 ± 0.6
10.6 ± 0.5
17.4 ± 0.4
28.3 ± 0.4
38.4 ± 0.4 Of Comparative Example 8
Porcine Placenta Extract
1.5 ± 0.8
5.2 ± 0.9
5.7 ± 0.2
13.2 ± 0.3
29.4 ± 0.8
Retinol
2.1 ± 0.8
5.3 ± 0.5
8.2 ± 0.3
15.2 ± 0.2
24.0 ± 0.2

Experimental Example 2. Wrinkle improvement effect and skin irritation evaluation of cosmetics containing placenta extract>

The wrinkle improvement effect of the cosmetic of this invention was evaluated through the practical use test. As the evaluation target, creams of Example 4, Example 8 and Comparative Examples 9 to 17 were used, and 20 to 40-60 year old women were divided into 11 groups. Right around 2 months. The wrinkle improvement effect of the subject was evaluated through visual observation, and the experimental results are described by the 5-point spot method in Table 2 below.

Condition
Wrinkle improvement
Cream of Example 4
4.8
Cream of Example 8
4.5
Cream of Comparative Example 9
2.4
Cream of Comparative Example 10
2.5
Cream of Comparative Example 11
2.7
Cream of Comparative Example 12
2.5
Cream of Comparative Example 13
2.3
Cream of Comparative Example 14
2.2
Cream of Comparative Example 15
3.5
Cream of Comparative Example 16
2.0
Cream of Comparative Example 17
1.5
5 points: Very good, 4 points: Excellent, 3 points: Normal, 2 points: Bad, 1 point: Very bad

The results of Table 2 showed that the cosmetic composition containing the placenta extract of the present invention showed a significantly higher antiwrinkle effect than the cosmetics of the other comparative examples, and no skin irritation was observed in all the subjects to which the cream of the present invention was applied.

Experimental Example 3. Evaluation of Flavor of Cosmetics Containing Horse Placenta Extract

Example 4 through the experimenters of Experimental Example 2 to evaluate the preference effect of the adsorption process using high-purity nitrogen and activated carbon to remove the inherent off-flavor of horse placenta extract when preparing the placenta extract of the present invention , And the flavor of the cream of Example 8 and Comparative Example 15 was compared and the experimental results are described in the following Table 3 by the five-point RBI method.

Condition
Preference for the aroma of cosmetics
Cream of Example 4
4.5
Cream of Example 8
2.5
Cream of Comparative Example 15
1.2
5 points: Very good, 4 points: Excellent, 3 points: Normal, 2 points: Bad, 1 point: Very bad

As a result of Table 2, cosmetics containing horse placenta extracts which had undergone adsorption process had better preference for fragrance than cosmetics containing horse placenta extracts which did not perform adsorption process. Cosmetics containing the extract was found to be more preferred because the inherent off-flavor of the horse placenta extract was removed than the cosmetics containing the placenta extract adsorbed under vacuum conditions.

Claims (6)

i) Primary proteolysis at 60 to 70 ° C. by adding 30 to 40 parts by weight of placenta to 0.2 to 0.6 parts by weight of protease selected from papain, protease, peptidase, pepsin or mixtures thereof and 65 to 100 parts by weight of water. Reacting;
ii) further adding 0.05 to 0.2 parts by weight of the protease to the reaction product of i) to perform a second proteolytic reaction at 60 to 70 ° C; And
iii) adding 1.0 to 1.5 parts by weight of activated carbon having a water content of 1 to 3% by weight to the reactant of ii) and then removing odor and color under high purity nitrogen or vacuum pressure of 0.1 to 0.5 atmosphere;
Wrinkle improvement composition characterized in that it comprises 0.0005 ~ 10% by weight of mal placenta extract is removed as an active ingredient. delete delete delete delete The method of claim 1,
The wrinkle improving composition is a lotion, cream, essence, cosmetic ointment, spray, cleansing foam, cleansing water, pack, body oil, lotion, foundation, lipstick, mascara, makeup base, sunscreen, makeup remover and cleanser from the group Wrinkle improvement composition, characterized in that any one of the selected formulation.

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