KR102381273B1 - Cosmetic composition with promoting collagen synthesis and shirinking skin-pores, and Manufacturing method thereof - Google Patents

Abstract

The present invention relates to cosmetics used in the manufacture of a cosmetic product having excellent functions of alleviating skin wrinkles, reducing skin pores, and maintaining skin elasticity, and more specifically, to cosmetics which are a cosmetic material that can promote collagen synthesis in the skin and reduce the pores to alleviate the skin wrinkles and the skin elasticity and a functional cosmetic product using the same.

Description

Cosmetic composition for promoting collagen synthesis and pore improvement, and manufacturing method thereof

The present invention relates to a cosmetic which is a cosmetic material capable of promoting collagen synthesis in the skin and reducing pores to improve skin wrinkles and skin elasticity, and a method for manufacturing the same.

The skin is composed of the epidermis, dermis, and subcutaneous tissue. Of these, the epidermis functions as a physical protective film to regulate body temperature and protects the body from the external environment, while the dermis is a connective tissue located between the epidermis and subcutaneous fat layer, providing flexibility, elasticity and tension to the skin. It provides structural support for the epidermis. In the skin connective tissue present in the dermis layer, there is collagen, an extracellular matrix protein, which is usually known to account for about 30% of the mammalian body protein. It consists of three amino acid residues: glycine, proline, and hydroxyproline. So far, 19 types of collagen have been studied, and there are Type I, Type III, and the rest of Type II and Type IV in adult skin, but Type I collagen is the most abundant in the dermal layer of the skin, and is the main protein that accounts for 90% It maintains the skin connective tissue and plays a role in giving strength and tension to the skin, and the newly synthesized procollagen is secreted into the extracellular space of skin cells through an enzymatic reaction to form collagen fibers.

Fibroblasts in the dermal layer are responsible for collagen expression, and as the age increases, the action and number of fibroblasts in the dermal layer decrease, resulting in a decrease in the synthesis of structural proteins such as collagen, and the breakdown of collagen. The amount of enzyme MMPs (Matrix MetalloProteases) increases, moisture in the skin cells is lost (Fisher, G. J et al., 2002), and the stratum corneum is also affected, resulting in changes in the skin structure. As we get older, the effects of UV rays accumulate, and as we reach a point where collagen degradation becomes dominant over collagen synthesis, the amount of collagen, which serves as a structure supporting the skin, decreases, leading to wrinkles.

Therefore, in the improvement of skin aging and wrinkles, it is important to find a safe material that promotes collagen synthesis and inhibits the action of collagenase, and various studies on changes in collagen in the skin are being conducted.

Polydeoxyribonucleotide (hereinafter referred to as 'PDRN'), which is recently used as a core material in cosmeceuticals and bio-cosmetics, is manufactured from DNA extracted from salmon semen, and a deoxyribonucleotide polymer of 50 to 2000 base pairs is used. contains PDRN is mainly used for the treatment and tissue repair of wounds caused by skin transplantation, but it is also used at the discretion of the medical staff for some tissue regeneration or inflammation treatment for which there is no suitable treatment. It is known to be used in the treatment of a wide range of disorders such as

In addition, it stimulates the A2 receptor, which is a skin regeneration signal transmitter, to promote secretion of various growth factors, generate capillaries by VEGF (vascular endothelial cell growth factor), improve blood circulation, anti-inflammatory action, and prevent capillary leakage. is known

It is being developed and commercialized as a skin improvement cosmetic with various functions such as wound treatment and normal organization using PDRN, acne scars, accident scars, stretch marks improvement, skin atrophy, fine wrinkles, acne skin, melasma, blemishes, etc. However, it is still an effective product Development is insufficient.

In addition, the low-molecular compound that promotes collagen production is a retinoid that is currently widely used to improve skin wrinkles, blemishes, and pigmentation. is being studied In addition to methods for promoting collagen synthesis, there are ultrasound treatment, laser dermabrasion, botulinum toxin injection, restylene injection, etc.

With the development of extensive research for the prevention and alleviation of such wrinkle formation, important functions of collagen in the skin are revealed, and research is underway to suppress skin wrinkles that occur with aging by developing raw materials that promote the synthesis of collagen in the skin. , various cosmetics are being developed and sold.

Korean Patent Registration No. 10-1722181 (published on March 31, 2017)

The present invention relates to a cosmetic comprising polydeoxyribonucleotide and botulinum toxin, known as collagen synthesis promoting ingredients, and an optimal composition for preventing skin problems caused by botulinum toxin, a method for manufacturing the same, and wrinkle improvement and pores using the same We aim to provide functional cosmetics for improvement.

Cosmetics of the present invention for solving the above problems include polydeoxyribonucleotides, botulinum toxin, and lidocaine.

The cosmetic of the present invention comprises a first step of preparing a first solution by mixing and stirring polydeoxyribonucleotide, botulinum toxin, a nonionic emulsifier and an aqueous ethanol solution; A second step of preparing a second solution by mixing and stirring the first solution and the natural plant complex extract in a weight ratio of 1: 0.3 to 0.5; and a third step of preparing a third solution by mixing and stirring the second solution, lidocaine, low molecular weight sodium hyaluronate, and an anionic emulsifier.

In addition, the present invention is to provide a cosmetic comprising the cosmetic.

The cosmetic of the present invention is excellent in the effect of promoting collagen synthesis in the skin and improving the pores through pore shrinkage. In addition, the cosmetic of the present invention containing a specific plant extract has excellent anti-inflammatory effect and skin soothing effect, and reduces skin troubles. It does not cause it, and by using it, various types of functional cosmetics can be manufactured.

Hereinafter, the present invention will be described in more detail through a method for manufacturing the cosmetic of the present invention.

The cosmetic of the present invention includes polydeoxyribonucleotide, botulinum toxin, and lidocaine.

The cosmetic of the present invention includes a first step of preparing a first solution by mixing and stirring polydeoxyribonucleotide, botulinum toxin, a nonionic surfactant and an aqueous ethanol solution; A second step of preparing a second solution by mixing and stirring the first solution and the natural plant complex extract; and a third step of preparing a third solution by mixing and stirring the second solution, lidocaine, low molecular weight sodium hyaluronate and anionic emulsifier;

The cosmetic of the present invention is an invention in which PDRN and botulinum toxin are introduced together for skin wrinkle improvement by PDRN and botulinum toxin for skin wrinkle improvement by promoting collagen synthesis, and the first solution in step 1 is PDRN, botulinum toxin , a nonionic emulsifier and an aqueous ethanol solution.

The polydeoxyribonucleotide (PDRN, polydeoxyribonucleotide) in step 1 is a cell tissue regeneration active substance, a substance that promotes self-renewal of damaged cells and tissues, and is a DNA fragment extracted from salmon germ cells. Adenosine receptors include A1, A2a, A2b, and A3. A2a plays an important role in inflammation, oxygen consumption, ischemia, cell growth, and angiogenesis, and PDRN acts as an agonist for A2a. It regenerates damaged tissues and has anti-inflammatory properties.

And, the content of polydeoxyribonucleotide in the first solution of step 1 is 8 to 15% by weight, preferably 10 to 15% by weight, more preferably 12.0 to 14.5% by weight of the total weight of the first solution, at this time , If the polydeoxyribonucleotide content is less than 8% by weight, the collagen synthesis promoting effect may be insufficient and the skin wrinkle improvement effect may be reduced. Therefore, it is recommended to use it within the above range.

In addition, the botulinum toxin in step 1 is a trade name for a muscle contraction injection of Allergan Inc., an American pharmaceutical company used for nerve and muscle diseases and wrinkle removal, etc. It is made by refining botulinum toxin, Toxin is a substance that has been used for cosmetic purposes such as removing wrinkles around the eyes since the 1990s. Botulinum toxin is a highly toxic neurotoxin excreted from a bacterium named Clostridium botulinum, and is a substance obtained by separating and purifying beneficial components from this dangerous toxin.

And, the content of botulinum toxin in the first solution of step 1 is about 0.01 to 0.05 wt%, preferably 0.02 to 0.04 wt%, at this time, if the botulinum content is less than 0.01 wt%, the amount of botulinum toxin is too small The effect of improving skin wrinkles and reducing pores may be insignificant, and if it is used in excess of 0.05% by weight, it is recommended to use it within the above range because problems such as skin troubles and skin tightness may occur.

In addition, the first solution of step 1 uses a nonionic surfactant for miscibility of PDRN and botulinum toxin, and the nonionic surfactant is ethylene glycol monostearate, glyceryl monostearate, glyceryl stearate, poly Glyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG-150 laurate, PEG-400 monolaurate, polyoxyethylene monolaurate, polysorbate, polyoxyethylene octylphenyl ether, PEG It may include at least one selected from polyoxyethylene tridecyl ether, polypropylene glycol butyl ether and stearoyl monoisopropanol amide, preferably ethylene glycol monostearate, glyceryl monostearate, glyceryl stearate, It may include at least one selected from PEG-150 laurate, PEG-400 monolaurate and PEG polyoxyethylene tridecyl ether, and more preferably, the nonionic surfactant is PEG-150 laurate and PEG-400. It may include at least one selected from monolaurate.

And, the content of the nonionic surfactant in the first solution in step 1 is 0.1 to 0.5% by weight, preferably 0.2 to 0.5% by weight, more preferably 0.3 to 0.5% by weight, and the nonionic surfactant content is 0.1% by weight If it is less than %, the miscibility of PDRN and botulinum toxin may be lowered, and using more than 0.5 wt% is an excessive use, which is uneconomical and may cause skin troubles due to the excessively used nonionic surfactant. it's good

In addition, the aqueous ethanol solution serves as a solvent for the cosmetic components used in steps 2 to 3 in addition to PDRN and botulinum toxin, and it is preferable to use an aqueous solution of ethanol having a concentration of 20 to 30% by volume. At this time, if the concentration of the aqueous ethanol solution is less than 20% by volume or exceeds 30% by volume, miscibility between cosmetic materials may be deteriorated due to a component that is not well dissolved in an aqueous ethanol solution, which is a solvent, during cosmetic preparation. And, the content of the aqueous ethanol solution in the first solution is 100% by weight of the remaining amount other than PDRN, botulinum toxin and nonionic surfactant described above.

Next, step 2 is a process of preparing a second solution by mixing and stirring the first solution prepared in step 1 and the natural plant complex extract.

The mixing ratio of the first solution and the natural plant complex extract is 1: 0.3 to 0.5 weight ratio, preferably 1: 0.35 to 0.40 weight ratio is appropriate. may be insufficient, and if it is used in excess of 0.5 weight ratio, there may be a problem that the rate of promoting collagen synthesis is rather reduced, so it is recommended to use it within the above range.

The natural plant complex extract relieves the toxicity of botulinum toxin in the first solution, and serves to impart functions such as skin whitening and antioxidant to cosmetics. It may be a hot water extract or a C ₁ to C ₄ lower alcohol extract of a natural plant mixture comprising the leaves and roots of dandelion, and comfrey leaves, preferably a hot water extract or an ethanol extract, more preferably ethanol (alcohol (alcohol) ) may be an extract.

Among the natural plant mixtures, Angelica is a perennial aromatic herb belonging to the Asteraceae and generally uses roots, and is one of representative herbal medicines. As the efficacy of Angelica keis, herbal gangmok is a plant known to have the effect of warming the inside, stopping pain, removing clumping blood, stopping vomiting, recovering from fatigue, diarrhea, abdominal pain, toothache, back pain, and uterine bleeding. Among the efficacy of Angelica quai, the representative effect is the blood circulation effect, which is known for the efficacy of decursin and decurcinol angelate, which are one of the main components of Angelica quai.

Among the natural plant mixtures, Ulmus davidiana is also called sugar oil (唐楡) and black oil (黑楡). It grows at the foot of a mountain or in a valley. The height is 15m, the diameter of the base is 50~70cm, the small branches have long soft hairs, and the bark is light gray. The leaves are alternate phyllotaxis and have an inverted oval shape or a long oval shape with a length of 4-8 cm. The tip of the leaf is gradually sharpened and the bottom is distorted, the edge has sharp double serrated teeth, the front side is slightly rough, and there are hairs on the veins on the back side. The petiole is 3-7mm long, and there are dense long soft hairs. As a raw material that has been used as a herbal medicine for a long time, in the present invention, it functions to prevent skin oxidation and keeps the skin clear and healthy.

The base paper of the natural plant mixture contains steroid-based lipid components, such as tenuigenin A, tenugenin B, polygalytol, tenuidine, and the like, and these components are antioxidant and anti-inflammatory in the skin. And it is known to help keep the state of mind and body clean by suppressing the secretion of components such as dopamine, which is a neurotransmitter.

Among the natural plant mixture, Adenophora Stricta Leaf & Root is used for the effect of preventing tissue damage due to skin inflammation and responding to biological tissue defense against various infectious agents.

Among the natural plant mixtures, comfrey (Symphytum officinale) is a dicotyledonous plant cultivated for medicinal or fodder use. In oriental medicine, the leaves and roots are used as a medicinal herb called Gamburi (甘富利). It is a plant that has been used as a medicine for gastric ulcer, anemia, boil, sore throat, and dermatitis.

The natural plant mixture may contain 10 to 15.0% by weight of raw paper, 20 to 32% by weight of dandelion leaves and roots, 5.0 to 10% by weight of comfrey leaves, and 100% by weight of the remaining amount, preferably raw paper 11.0 to 14.0% by weight, 24 to 30% by weight of Dandelion leaves and roots, 5.0 to 8.5% by weight of comfrey leaves, and 100% by weight of the remaining amount of Angelica, more preferably, 12.0 to 14.0% by weight of raw paper, Danangdae leaf and 25.0 to 29.5% by weight of the root, 5.5 to 8.0% by weight of the comfrey leaf, and 100% by weight of the remaining amount of Angelica, without causing skin troubles, is advantageous in functional aspects such as skin wrinkle improvement effect and skin whitening. In this case, the weight % of the plant in the natural plant mixture is based on dry weight.

For a preferred embodiment of preparing the natural plant complex extract, dried Angelica keis, dried elm root, dried raw paper, dried dandelion leaves and roots and dried comfrey leaves were prepared, respectively, and then they were mixed Step 2-1 to prepare the mixture; A second step of extracting an active ingredient beneficial to the skin by supporting the mixture in an aqueous ethanol solution having a concentration of 70 to 80% by volume, and then supporting the mixture for 4 to 6 hours at 15 to 35°C; A third step of first filtering the extract obtained by performing step 2 to obtain a first filtrate; Step 4 of preparing a secondary filtrate by adding and stirring 1,2-hexanediol and ethylhexylglycerin to the first filtrate, followed by secondary filtering; Step 5 of concentrating and filtering the second filtrate under reduced pressure to obtain a concentrate under reduced pressure; And step 6 of filtering the concentrate under reduced pressure; It can be manufactured by performing the process.

The concentration under reduced pressure in the five steps of the natural plant complex extract manufacturing process can be performed through a general method used in the art, and is performed to remove solvents and impurities.

And, the filtering of steps 3, 4, and 6 of the natural plant complex extract manufacturing process can be performed through a general filtering method used in the art.

Next, in the cosmetic manufacturing process of the present invention, step 3 is a process of preparing a third solution by mixing and stirring the second solution prepared in step 2 with lidocaine, low molecular weight sodium hyaluronate and anionic emulsifier.

The lidocaine is a substance used as a local anesthetic, and in the present invention serves to assist in skin penetration, absorption and delivery of skin improving ingredients such as polydeoxyribonucleotides, botulinum toxin, and plant extracts. The amount of lidocaine used is preferably 0.001 to 0.100 parts by weight, preferably 0.005 to 0.050 parts by weight, more preferably 0.005 to 0.030 parts by weight, based on 100 parts by weight of the second solution, in which case, the amount of lidocaine used is 0.01 parts by weight. If it is less than, the amount used is too small, and the effect of increasing the skin penetration, absorption and delivery of the skin improvement ingredient may be insufficient. Therefore, it is recommended to use it within the above range.

The low molecular weight sodium hyaluronate (Sodium Hyaluronate) serves as a moisturizing component and a dispersant in the cosmetic composition, and has a weight average molecular weight of 10,000 to 200,000 Daltons, preferably a weight average molecular weight of 20,000 to 160,000 Daltons, more preferably a weight average molecular weight of 25,000 to It is recommended to use 150,000 Daltons. And, the amount of low molecular weight sodium hyaluronate used is preferably 1.0 to 5.0 parts by weight, preferably 1.5 to 4.0 parts by weight, more preferably 1.8 to 3.5 parts by weight, based on 100 parts by weight of the second solution. At this time, if the amount of low molecular weight sodium hyaluronate used is less than 1.0 parts by weight, the dispersion and storage stability of the cosmetic may be deteriorated and precipitates may occur during long-term storage. There may be issues with this occurring.

In addition, the anionic surfactant serves to increase the miscibility between the second solution component and lidocaine and low molecular weight sodium hyaluronate, and a general anionic surfactant used in the art may be used, preferably a long-chain alkyl alkylarylsulfonates such as sodium, potassium, ammonium and sodium dodecylbenzene sulfonate of sulfonic acid salts; dialkyl sodium sulfosuccinates such as sodium dodecylbenzenesulfonate; dialkyl sodium sulfosuccinates such as sodium bis-(2-ethylthioxyl)-sulfosuccinate; and at least one selected from alkyl sulfates such as sodium lauryl sulfate. And, the amount of the anionic surfactant used is 0.05 to 0.50 parts by weight, preferably 0.05 to 0.40 parts by weight, more preferably 0.12 to 0.35 parts by weight, based on 100 parts by weight of the second solution, and the amount used is 0.05 If it is less than 1 part by weight, there may be a problem of generating solids due to insufficient miscibility between cosmetic components, and if it is used in excess of 0.50 parts by weight, it is excessive use and may cause skin troubles, so it is recommended to use it within the above range.

In addition, in step 3, a cosmetic may be prepared by adding additives such as a thickener and an emulsion stabilizer in addition to lidocaine, low molecular weight sodium hyaluronate, and anionic surfactant.

As the thickener, at least one selected from Polyquaternium-10 and Guar Hydroxypropyltrimonium Chloride may be used, and it may be used when the viscosity of the cosmetic is manufactured too low.

In addition, the emulsion stabilizer may use a general emulsion stabilizer used in the art, preferably, a small amount of carbomer may be used, and more preferably, 0.1 to 0.3 parts by weight based on 100 parts by weight of the second solution. An additional degree can be used.

In step 3, the second solution, lidocaine, low molecular weight sodium hyaluronate, anionic surfactant, or the second solution, lidocaine, low molecular weight sodium hyaluronate, anionic surfactant and the above additives can be mixed to prepare a cosmetic, Preferably, the cosmetic can be prepared by emulsifying it using a high-pressure liquid emulsification method.

For an example of the high pressure liquid emulsification method, for 10 to 30 minutes, preferably 10 to 20 by a high pressure liquid emulsification method using a high pressure liquid emulsifier (microfluidizer) under 800 to 1,200 bar pressure, preferably 800 to 1,000 bar pressure. It is carried out for minutes, and the active ingredient particles are pulverized through this method to prepare a cosmetic in which the ingredients such as the second solution, lidocaine, and low molecular weight sodium hyaluronate are emulsified evenly. And, when performing the high-pressure liquid emulsification method, at this time, the temperature inside the ultra-high pressure disperser of the high-pressure liquid emulsifier is 45 ~ 60 ℃, it is preferable to carry out under the condition that the outlet temperature of the ultra-high pressure disperser is about 10 ~ 15 ℃.

The cosmetic of the present invention prepared in this way has an excellent effect of promoting collagen synthesis in the skin, pore improvement effect through pore shrinkage, excellent anti-inflammatory and skin soothing effect, does not cause skin troubles, and uses it in various forms of functional cosmetics can be manufactured.

The cosmetic of the present invention is a cream, foam, nourishing lotion, flexible lotion, pack, soft water, emulsion, makeup base, essence, soap, liquid detergent, bath agent, sunscreen cream, sun oil, suspension, emulsion, paste, gel, It can be prepared as cosmetics of various formulations such as lotions, powders, sprays, and the like, but is not limited thereto.

By applying a cosmetically acceptable carrier to the cosmetic of the present invention, for example, oil, water, surfactant, humectant, lower alcohol, thickener, chelating agent, colorant, preservative, fragrance, etc. can be appropriately blended to manufacture cosmetics. there is.

When the cosmetic formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide as a carrier component and the like may be used, but is not limited thereto. These may be used alone or in combination of two or more.

In addition, when the cosmetic formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component, and in particular, in the case of a spray, additional chloro propellants such as, but not limited to, fluorohydrocarbons, propane/butane or dimethyl ether. These may be used alone or in combination of two or more.

In addition, when the cosmetic formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier may be used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol , benzyl benzoate, propylene glycol, 1,3-butylglycol oil and the like can be used, in particular cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil, glycerol aliphatic esters, polyethylene glycol and / Or a fatty acid ester of sorbitan may be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the cosmetic formulation of the present invention is a suspension, as a carrier component a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used, but is not limited thereto. These may be used alone or in combination of two or more.

Hereinafter, the present invention will be described in more detail based on examples. However, the following examples are provided to aid understanding of the present invention, and should not be construed as limiting the scope of the present invention to the following examples.

[Example]

Preparation Example 1: Preparation of natural plant complex extract

A mixture was prepared by preparing dried Angelicasis, dried Angelica root, dried raw paper, dried Angelica root and dried comfrey leaves, respectively, and then mixing them. The mixture contains Angelica oleifera root 23.5% by weight, raw paper 12.8% by weight, Dandelion leaves and roots 28.4% by weight, comfrey leaves 6.2% by weight, and the remaining balance of 100% by weight.

Next, the mixture was supported in an aqueous solution of ethanol at a concentration of 75 to 76% by volume, and then the skin beneficial active ingredients were brewed at about 30° C. for 5 hours. Then, the obtained extract was filtered with a filter membrane having an average pore size of 25 μm to obtain a primary filtrate.

Next, with respect to 100 parts by weight of the first filtrate, 5.5 parts by weight of 1,2-hexanediol and 0.2 parts by weight of ethylhexyl glycerin were added to 94.9 ml, and then sufficiently stirred, and the stirred solution had an average pore size of 10 μm Secondary filtration was performed with a filter membrane to obtain a secondary filtrate.

Next, the secondary filtrate was concentrated under reduced pressure, followed by filtration to obtain a concentrate under reduced pressure.

Next, the reduced pressure concentrate was filtered with a filter membrane having an average pore size of 10 μm to obtain a natural plant complex extract.

Comparative Preparation Example 1

After preparing a mixture by mixing 40.5 wt % of dried Angelica root and 59.5 wt % of dried Angelica root, the mixture was prepared in the same manner as in Preparation Example 1, and the first filtrate, the second filtrate, and the reduced pressure concentrate were sequentially prepared. After obtaining, it was filtered to obtain a natural plant complex extract.

Example 1: Preparation of cosmetics

Polydeoxyribonucleotide (PDRN) 13.2% by weight, botulinum toxin 0.03% by weight, glyceryl monostearate, and PEG polyoxyethylene tridecylether in a 1:0.2 weight ratio of 0.42% by weight of a nonionic surfactant mixed with the remaining amount After adding an aqueous ethanol solution (concentration of 25% by volume) to the reactor, the first solution was prepared by slowly stirring at about 25°C.

Next, the first solution and the natural plant complex extract of Preparation Example 1 prepared above were mixed in a weight ratio of 1:0.40, and then gently stirred at about 25° C. to prepare a second solution.

Next, based on 100 parts by weight of the second solution, 0.015 parts by weight of lidocaine, 2.1 parts by weight of low molecular weight sodium hyaluronate having a weight average molecular weight of 50,000 to 100,000 daltons, sodium bis-(2-ethylthioxyl)-sulfosuccine as an anionic surfactant 0.18 parts by weight of cinate and 0.12 parts by weight of Carbomer as an emulsion stabilizer were put into a high-pressure liquid emulsifier, and then emulsified in a liquid phase under a pressure of about 850 bar for 15 minutes to prepare an emulsified cosmetic. The internal temperature of the ultra-high pressure disperser of the high-pressure liquid emulsifier was 48-50°C, and the outlet temperature of the ultra-high-pressure disperser was about 10-12°C.

Examples 2 to 4

Cosmetics were prepared in the same manner as in Example 1, but as shown in Table 1 below, cosmetics were prepared by varying the amount of the composition used for preparing the cosmetics, and Examples 2 to 4 were carried out, respectively.

Comparative Example 1

Cosmetics were prepared in the same composition, content and method as in Example 1, but using the natural plant complex extract prepared in Comparative Preparation Example 1 instead of the natural plant complex extract of Preparation Example 1 was used.

Comparative Example 2 ~ Comparative Example 4

Cosmetics were prepared in the same manner as in Example 1, but as shown in Table 1 below, cosmetics were prepared by varying the amount of composition used for preparing cosmetics, and Comparative Examples 2 to 4 were carried out, respectively.

division Example 1 Example 2 Example 3 Example 4 first solution PDRN 13.2% by weight 15.0 wt% 11.3% by weight 13.2% by weight botulinum toxin 0.03 wt% 0.02% by weight 0.04 wt% 0.03 wt% nonionic
Surfactants 0.42% by weight 0.48% by weight 0.40% by weight 0.42% by weight ethanol aqueous solution Remaining balance 100% by weight second solution 1st solution: natural plant complex extract weight ratio 1:0.40
(Preparation example 1) 1:0.40
(Preparation example 1) 1:0.40
(Preparation example 1) 1:0.40
(Preparation example 1) third solution Lidocaine ⁽¹⁾ 0.015 parts by weight 0.015 parts by weight 0.015 parts by weight 0.045 parts by weight (1) parts by weight based on 100 parts by weight of the second solution

division Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 first solution PDRN 13.2% by weight 18.0 wt% 13.0% by weight 13.2% by weight 13.2% by weight botulinum toxin 0.03 wt% 0.002% by weight 0.07% by weight 0.03 wt% 0.03 wt% nonionic
Surfactants 0.42% by weight 0.48% by weight 0.40% by weight 0.42% by weight 0.42% by weight ethanol aqueous solution Remaining balance 100% by weight second solution 1st solution: weight ratio of natural plant complex extract 1:0.40
(Comparative preparation example 1) 1:0.40
(Preparation example 1) 1:0.40
(Preparation example 1) 1:0.40
(Preparation example 1) 1:0.10
(Preparation example 1) third solution Lidocaine ⁽¹⁾ 0.015 parts by weight 0.015 parts by weight 0.015 parts by weight 0.115 parts by weight 0.015 parts by weight (1) parts by weight based on 100 parts by weight of the second solution

Experimental Example 1: Skin cytotoxicity test

RAW 264.7 cells (Korea Cell Line Bank) were suspended in DMEM (Gibco) medium supplemented with 10% fetal bovine serum at a concentration of 2×10 ⁵ cells/mL, and 200 μl each in 96-well plates (96-well). plate) was inoculated and attached. Thereafter, the cosmetics prepared in Examples 1 to 4 and Comparative Examples 1 to 4 were treated at a concentration of 1000 μg/ml, respectively, and cultured for 20 hours. After adding 20 μl of a 5 mg/ml MTT solution per well, the culture was further incubated for 4 hours. After removing the supernatant, 100 μl of DMSO was added, and then absorbance was measured at 570 nm. Cell viability was calculated according to the following formula with the PBS-treated negative control group being 100%, and the results are shown in Table 3 below.

[Equation 1]

Cell viability (%) = (absorbance value of cosmetic treatment group / absorbance value of negative control group) × 100%

division Cell viability (%) negative control 100 Example 1 99.8 Example 2 100.3 Example 3 99.6 Example 4 99.2 Comparative Example 1 95.3 Comparative Example 2 101.4 Comparative Example 3 93.8 Comparative Example 4 98.1 Comparative Example 5 94.7

Looking at the skin cytotoxicity test results in Table 3, Examples 1 to 4 and Comparative Example 2 showed almost no toxicity to skin cells with a cell viability of 99.0% or more.

In contrast, Comparative Example 1 using the natural plant complex extract of Comparative Preparation Example 1 showed an increase in cytotoxicity as compared to Example, and also, when preparing the second solution, with respect to the first solution, natural plants Comparative Example 5 using the complex extract in an amount of less than 0.30 weight ratio also showed a sharp decrease in cell viability when compared with Examples. Through this, it was confirmed that the natural plant complex extract has the effect of reducing skin cytotoxicity caused by botulinum toxin.

In addition, in the case of Comparative Example 3, in which the botulinum toxin in the first solution was used in excess of 0.05% by weight, when compared to Example 3, the cytotoxicity rapidly increased, and also, when the third solution was prepared, the second solution 100 Comparative Example 4 in which lidocaine was used in excess of 0.100 parts by weight based on parts by weight also showed a somewhat increased cytotoxicity compared to Example 4.

Experimental Example 2: Evaluation of the ability to inhibit elastase activity and collagenase activity

(1) Elastase activity inhibitory ability

Elastase is an enzyme that breaks down elastin, an important substrate for maintaining skin elasticity in the dermis. Therefore, it is possible to suppress skin aging by lowering the activity of elastase, which is one of the main causes of skin aging.

In order to confirm the skin wrinkle improvement effect of the cosmetics prepared in Examples and Comparative Examples, inhibition of elastase activity was measured in the following manner, and the results are shown in Table 4 below.

For the measurement method, 100 μl of 100 mM Tris-HCl buffer (pH 8.0) was dispensed in a 96-well plate, the cosmetic was added in the same amount, and then 20 μl of 2.9 mM N-Succinyl-(Ala)3-pnitroanilide was dispensed. . Then, 15 μl of the elastase enzyme solution prepared in 0.2 unit was added, mixed, and reacted at 25° C. for 20 minutes. The reaction was terminated by leaving the reactant on ice for 5 minutes, and the absorbance was measured using a microplate reader having a wavelength of 410 nm, and the elastase activity rate was measured by substituting it in Equation 1 below, and the maximum inhibitory activity of tyrosinase. Concentrations that could represent 50% (IC ₅₀ ) were calculated (Table 5). In this case, the positive control group used a reactant to which ursolic acid was added, and distilled water was used as the negative control group.

[Equation 1]

Elastase activity rate (%) = 1-{(Sample solution absorbance - negative control absorbance) / negative control absorbance × 100 (%)}

(2) Evaluation of the ability to inhibit collagenase activity

Collagenase is an enzyme that decomposes collagen, which is an important substrate for inhibiting skin wrinkles in the dermis and maintaining elasticity. Therefore, secretion of collagenase accelerates skin aging and causes wrinkles.

To evaluate the inhibition of collagenase activity, 4 nM CaCl ₂ was added to the reaction well of 0.1 M tris-HCl buffer (pH 7.5) and 4-phenylazobenzyloxycarbonyl-Pro-LeuGly-Pro-D-Arg (0.3mg/mL) was dissolved in a substrate solution. To a mixture of 0.25 mL and 0.1 mL of the sample solution, 0.15 mL of collagenase (0.2mg/mL, Sigma Co) was added, left at room temperature for 20 minutes, and then 0.5mL of 6 wt% citric acid was added. After stopping the reaction, 1.5 mL of ethyl acetate was added to measure the absorbance at 320 nm, and the results are shown in Table 4 below.

division Elastase activity inhibitory ability
(IC ₅₀ =mg/ml) collagenase
Activity inhibition rate (%) positive control
(Ursolic acid) 56.85 - positive control
(Ascorbic acid 0.05%) - 71.3 Example 1 54.91 72.9 Example 2 53.93 72.0 Example 3 56.96 70.6 Example 4 58.85 72.4 Comparative Example 1 57.37 60.3 Comparative Example 2 59.93 69.5 Comparative Example 3 53.90 74.1 Comparative Example 4 54.17 72.2 Comparative Example 5 62.73 58.2

Looking at the elastase inhibitory ability measurement results in Table 4, it was confirmed that Examples 1 to 4 had IC ₅₀ at a generally low concentration when compared to the overall positive control group. It was confirmed that the ability to inhibit enzyme activity was excellent.

In contrast, Comparative Example 1 using the extract of Comparative Preparation Example 1 had a skin cytotoxicity problem, as confirmed in Experimental Example 1, and had a higher IC ₅₀ concentration than Example 1, but showed a high elastase activity inhibition ability, The inhibition rate of collagenase activity showed a rather low result.

And, Comparative Example 2 using 18% by weight of PDRN in the first solution and using less than 0.01% by weight of botulinum toxin Examples 1 to 3 using 11.0 to 15.0% by weight despite the high use of PDRN at 18% by weight Compared with , the inhibition of elastase activity and the inhibition rate of collagenase activity were low.

Comparative Example 3, in which botulinum toxin in the first solution was used in excess of 0.05% by weight, has excellent elastase activity inhibitory ability and collagenase activity inhibitory effect, but has a problem of skin cytotoxicity as confirmed in Experimental Example 1.

In addition, Comparative Example 4, in which lidocaine was used in excess of 0.100 parts by weight with respect to 100 parts by weight of the second solution when preparing the third solution, was also excellent in inhibiting elastase activity compared to Example 4, but the effect of increasing the inhibition of collagenase activity was no longer, and in Experimental Example 1, only the problem showing the result of a rather increased cytotoxicity was derived.

In addition, when preparing the second solution, Comparative Example 5 using the natural plant complex extract in an amount of less than 0.30 weight ratio to the first solution has a good collagenase activity inhibition rate, but the elastase activity inhibition ability is sharp compared to other examples. showed significantly lower results.

Through Experimental Example 2, it was confirmed that the cosmetic of the present invention has excellent elastase activity inhibitory ability and collagenase activity inhibitory effect, and through this, it is suitable for use as a cosmetic for cosmetics having excellent anti-wrinkle and skin pore-reducing functions. was able to confirm

Experimental Example 3: Experiment with skin whitening effect through tyrosinase inhibitory effect

In order to analyze the tyrosinase inhibitory effect of the cosmetics prepared in Examples 1 to 3, Comparative Examples 1 and 5, the cosmetic was prepared by converting L-tyrosine to L- using mushroom-derived tyrosinase (Sigma). The efficacy of inhibiting conversion to DOPA (L-3,4-dihydroxy phenylalanine) was analyzed.

For the tyrosinase inhibition assay for L-tyrosine, 120 μl 0.1 M sodium phosphate buffer (pH 6.5), 20 μl 1,000 unit/mL tyrosinase, and 20 μl samples were added to each 96-well microplate, respectively, for 10 minutes at room temperature. After the reaction, 40 μl 1.5 mM L-tyrosine was added, and absorbance was measured at 490 nm at 25° C. for 10 minutes.

Distilled water was used for the negative control group (N-control), the positive control group (P-control) was kojic acid (KA, Sigma), and the experimental group was analyzed by 20, 40, 80, 100 μg/ml concentration as cosmetic content, and the measured IC The values of ₅₀ (half maximal inhibitory concentration) are shown in Table 5 below.

division Tyrosinase inhibitory ability
(IC ₅₀ =mg/ml) positive control 50.78 Example 1 55.44 Example 2 54.31 Example 3 55.40 Comparative Example 1 63.54 Comparative Example 5 61.92

Looking at the tyrosinase inhibitory activity measurement results in Table 5, Examples 1 to 3 showed a lower skin whitening effect than the positive control group, but it was confirmed that they had an IC ₅₀ at a concentration of 60 mg/ml or less, and this result was compared In Comparative Examples 1 and 2 using the natural plant complex extract of Preparation Example 1, when compared with Comparative Example 5 in which the natural plant complex extract was used in an amount of less than 0.30 weight ratio, it showed a relatively very high tyrosinase inhibitory ability.

Through Experimental Example 2, it was confirmed that the cosmetic of the present invention has excellent skin whitening effect, and through this, it can be confirmed that it can be used as a cosmetic for functional cosmetics having excellent skin wrinkle improvement and skin pore reduction, as well as skin whitening effect. there was.

Comparative Example 6: Preparation of cosmetics

Cosmetics were prepared in the same manner as in Example 1, but cosmetics were prepared without using anionic surfactants and emulsion stabilizers when preparing the third solution.

Experimental Example 4: Storage stability (long-term dispersion stability) experiment of cosmetics

After shaking the cosmetics prepared in Examples 1 to 4 and Comparative Example 6, the absorbance (350 nm) was measured with a spectrophotometer for turbidity after being left at 30° C. for 1 hour and 48 hours, and the results are shown in Table 6 below. indicated. The turbidity change rate was measured based on Equation 2 below.

[Equation 2]

Turbidity change rate = 100%-(absorbance after 48 hours / absorbance after 1 hour) × 100%

division Absorbance after 1 hour Absorbance after 48 hours rate of change (%) Example 1 0.1892 0.1759 7.03 Example 2 0.1987 0.1882 5.28 Example 3 0.1730 0.1673 3.29 Example 4 0.1923 0.1849 3.84 Comparative Example 6 0.1721 0.1347 21.73

Looking at the turbidity change rate in Table 6, Examples 1 to 4 showed a turbidity change rate of less than 10% and cosmetic ingredients maintained high dispersibility, whereas Comparative Example 6 showed a turbidity change rate of 20% or more, and the storage container bottom Precipitation started to occur.

Through this, it was confirmed that the cosmetic of the present invention has excellent storage stability or long-term dispersion stability.

Claims (3)

Based on 100 parts by weight of the second solution containing the first solution and the natural plant complex extract in a weight ratio of 1:0.3 to 0.5, lidocaine 0.001 to 0.100 parts by weight, and low molecular weight sodium hyaluronate 1.0 to 5.0 parts by weight of 10,000 to 200,000 daltons of weight average molecular weight parts and 0.05 to 0.50 parts by weight of anionic surfactant,
The first solution contains 8 to 15% by weight of polydeoxyribonucleotide, 0.01 to 0.05% by weight of botulinum toxin, 0.1 to 1.0% by weight of a nonionic surfactant, and 20 to 30 of the remaining amount of the total weight It contains an aqueous solution of ethanol at a concentration of volume %,
The natural plant complex extract is an ethanol extract of a natural plant mixture comprising Angelica asiatica, Angelica elm root, raw paper, Angelica leaves and roots, and comfrey leaves,
The nonionic surfactant is ethylene glycol monostearate, glyceryl monostearate, glyceryl stearate, polyglyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG-150 laurate, PEG- 400 monolaurate, polyoxyethylene monolaurate, polysorbate, polyoxyethylene octylphenyl ether, PEG polyoxyethylene tridecyl ether, polypropylene glycol butyl ether and at least one selected from stearoyl monoisopropanol amide and
The anionic surfactants include, but are not limited to, alkylarylsulfonates such as sodium, potassium, ammonium and sodium dodecylbenzenesulfonate of long-chain alkylsulfonic acid salts; dialkyl sodium sulfosuccinates such as sodium dodecylbenzenesulfonate; dialkyl sodium sulfosuccinates such as sodium bis-(2-ethylthioxyl)-sulfosuccinate; and at least one selected from alkyl sulfates such as sodium lauryl sulfate.
delete 8 to 15% by weight of polydeoxyribonucleotide, 0.01 to 0.05% by weight of botulinum toxin, 0.1 to 1.0% by weight of a nonionic surfactant, and an aqueous ethanol solution having a concentration of 20 to 30% by volume of the remainder of the total weight are mixed and stirred to prepare 1 step of preparing a solution;
A second step of preparing a second solution by mixing and stirring the first solution and the natural plant complex extract in a weight ratio of 1: 0.3 to 0.5;
With respect to 100 parts by weight of the second solution, 0.001 to 0.100 parts by weight of lidocaine, 1.0 to 5.0 parts by weight of low molecular weight sodium hyaluronate having a weight average molecular weight of 10,000 to 200,000 Daltons, and 0.05 to 0.50 parts by weight of an anionic surfactant are mixed and stirred to obtain the third solution 3 steps of producing a process comprising;
The nonionic surfactant is ethylene glycol monostearate, glyceryl monostearate, glyceryl stearate, polyglyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG-150 laurate, PEG- 400 monolaurate, polyoxyethylene monolaurate, polysorbate, polyoxyethylene octylphenyl ether, PEG polyoxyethylene tridecyl ether, polypropylene glycol butyl ether and at least one selected from stearoyl monoisopropanol amide and
The anionic surfactants include, but are not limited to, alkylarylsulfonates such as sodium, potassium, ammonium and sodium dodecylbenzenesulfonate of long-chain alkylsulfonic acid salts; dialkyl sodium sulfosuccinates such as sodium dodecylbenzenesulfonate; dialkyl sodium sulfosuccinates such as sodium bis-(2-ethylthioxyl)-sulfosuccinate; and at least one selected from alkyl sulfates such as sodium lauryl sulfate,
The natural plant complex extract of step 2 is
Step 2-1 of preparing a mixture of dried Angelica keis, dried Angelica root, dried raw paper, dried Angelica basilica leaves and roots, and dried comfrey leaves, respectively, and then preparing a mixture; A second step of extracting an active ingredient beneficial to the skin by supporting the mixture in an aqueous ethanol solution having a concentration of 70 to 80% by volume, and then supporting the mixture for 4 to 6 hours at 15 to 35°C; A third step of first filtering the extract obtained by performing step 2 to obtain a first filtrate; Step 4 of preparing a secondary filtrate by adding and stirring 1,2-hexanediol and ethylhexylglycerin to the first filtrate, followed by secondary filtering; Step 5 of concentrating and filtering the second filtrate under reduced pressure to obtain a concentrate under reduced pressure; And step 6 of filtering the concentrate under reduced pressure; A method of manufacturing a cosmetic for promoting collagen synthesis and improving pores, characterized in that it is manufactured by performing the process.