Hydrolyzed sponge impregnated with peptide and cosmetic compositions comprising the hydrolyzed sponge

Abstract

The present invention relates to a hydrolyzed sponge impregnated with peptides and a cosmetic composition containing the same. The peptide-impregnated microneedles may have excellent skin affinity and skin penetration power, and when the microneedles penetrate the skin, the compressive force at the interface formed between the microneedles and the skin is strong, which can tighten pores present in the skin, such as pores, thereby allowing the peptides to be delivered to the skin more effectively. Accordingly, a cosmetic composition containing peptide-impregnated microneedles according to the present invention may have an excellent effect on improving skin wrinkles.

Description

Technology Field

[0001] The present invention relates to a hydrolyzed sponge impregnated with peptides and a cosmetic composition containing the same. Specifically, it relates to a hydrolyzed sponge that can provide an excellent skin wrinkle improvement effect by significantly improving the amount of peptide delivered to the skin by impregnating the hydrolyzed sponge with peptides, and a cosmetic composition containing the same. Background Technology

[0002] As modern people's interest in appearance and beauty increases, the cosmetics industry is growing rapidly day by day. In line with this trend, extensive research has been conducted on ingredients beneficial to the skin, enabling consumers to select cosmetics containing effective ingredients tailored to their individual needs or preferences.

[0003] The skin serves as the human body's primary defense organ against various harmful substances and is largely composed of three layers: the epidermis, dermis, and subcutaneous tissue. Consequently, even if functional ingredients are included in cosmetic compositions, there are limitations to their full efficacy if they cannot penetrate the outer layer of the skin. Therefore, research is needed to deliver active ingredients deep into the skin.

[0004] To this end, research has been conducted to reduce the size of emulsion particles when using emulsion formulations in cosmetic compositions, or to form liposomes, which are compositions similar to the skin, and deliver active ingredients deep into the skin by encapsulating them inside the liposomes. However, there are still limitations in improving the delivery of active ingredients because, when a cosmetic composition is applied to the outer surface of the skin, the particles of the composition still have to overcome the outer layer of the skin, such as dead skin cells.

[0005] Accordingly, the inventors have completed the present invention by impregnating naturally derived microneedles, such as hydrolyzed sponges, with peptides, which are skin-active ingredients, and delivering the peptides directly to the skin through the microneedles to provide an excellent skin wrinkle improvement effect. Prior art literature

[0006] (Patent Document 0001) KR 10-2022-0152218 A The problem to be solved

[0007] The present invention aims to provide a hydrolyzed sponge impregnated with peptides that has high skin affinity, excellent skin penetration, and can provide an excellent wrinkle improvement effect, and a cosmetic composition containing the same.

[0008] The technical problems of the present invention are not limited to those mentioned above, and other unmentioned technical problems will be clearly understood by those skilled in the art from the description below. means of solving the problem

[0009] According to an embodiment of the present invention, a peptide-impregnated microneedle is provided.

[0010] In addition, the microneedles may include a hydrolyzed sponge.

[0011] In addition, the peptide may include at least one of acetylated peptides; palmitoyl-based peptides; peptide copper complexes; polypeptides; and carnosine.

[0012] In addition, the peptide may include at least one of acetyl hexapeptide-8, ascorbic acid polypeptide, acetyl tetrapeptide-2, acetyl tetrapeptide-3, acetyl tetrapeptide-5, acetyl tetrapeptide-9, acetyl octapeptide-3, palmitoyl tripeptide-1, palmitoyl tripeptide-5, palmitoyl pentapeptide-4, nonapeptide-1, carnosine, and copper tripeptide-1.

[0013] In addition, the above peptide may include one contained inside a liposome.

[0014] In addition, the above peptide can be impregnated into microneedles at 45 to 55°C.

[0015] In addition, the peptide can be stirred for 4 to 5 hours and impregnated into microneedles.

[0016] In addition, the above peptide can be impregnated into microneedles at 0.04 to 0.07 MPa.

[0017] In addition, the peptide may be included in an amount of 0.01 to 0.5 weight% based on the total weight of the microneedle.

[0018] In addition, the above microneedles may be characterized by being treated with plasma.

[0019] In addition, the above plasma may be characterized as being an atmospheric pressure low-temperature plasma.

[0020] In addition, the plasma treatment can be performed before or after the peptide is impregnated into the microneedle.

[0021] According to another embodiment of the present invention, a cosmetic composition comprising the microneedles is provided.

[0022] In addition, the above cosmetic composition can provide an excellent skin wrinkle improvement effect. Effects of the invention

[0023] According to the present invention, microneedles impregnated with peptides may have excellent skin affinity and skin penetration capabilities. Furthermore, when the microneedles penetrate the skin, the compressive force at the interface formed between the microneedles and the skin is strong, so that pores and other openings in the skin can be tightened, allowing the peptides to be delivered to the skin more effectively and preventing other foreign substances from entering through the interface of the microneedles. Additionally, after the microneedles that have penetrated the skin are removed, surrounding cells quickly fill the space where they were removed, allowing the skin to regenerate.

[0024] Accordingly, a cosmetic composition comprising microneedles impregnated with peptides according to the present invention can have an excellent effect of improving skin wrinkles, giving elasticity to the skin, and producing a skin moisturizing effect. Brief explanation of the drawing

[0025] A brief description of each drawing is provided to help to better understand the drawings cited in the detailed description of the invention. Figure 1a shows a photograph of Example 1 taken under a microscope. Figure 1b shows a photograph of a microneedle taken under a microscope before it is impregnated with a peptide. Figure 2 shows a schematic diagram of the skin penetration device used in the Franz infusion cell assay experiment. Figure 3 shows photographs of the microneedle powder, Example 1, and Comparative Example 2 before peptide impregnation. Specific details for implementing the invention

[0026] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by a skilled expert in the art to which the present invention pertains. In general, the nomenclature used herein is well known and commonly used in the art. Furthermore, in describing embodiments of the present invention, detailed descriptions of related known components or functions are omitted if it is determined that such detailed descriptions would hinder understanding of the embodiments of the present invention. Additionally, while embodiments of the present invention will be described below, the technical concept of the present invention is not limited or restricted thereto and can be modified and implemented in various ways by those skilled in the art.

[0027] In this specification, when a part is described as including a certain component, it means that, unless specifically stated otherwise, it does not exclude other components but may include additional components. In this specification, the term "and / or" includes a combination of a plurality of related items or any one of a plurality of related items.

[0029] According to an embodiment of the present invention, a peptide-impregnated microneedle is provided.

[0030] The “peptide” of the present invention is a short chain of amino acids connected by peptide bonds, which can exhibit efficacy in improving wrinkles and elasticity by activating tissue growth factors and promoting collagen production. The peptide of the present invention can exhibit effects of improving skin wrinkles and enhancing skin elasticity by promoting collagen production, and can exhibit skin moisturizing effects by forming moisturizing proteins.

[0031] In one embodiment, the peptide may be in the form of a peptide complex. The “peptide complex” may refer to a component or structure in which a single peptide or two or more different peptide components are closely bonded physically or chemically.

[0032] In one embodiment, the peptide may be contained inside a liposome. When the peptide is contained inside a liposome, the amount of peptide delivered to the skin may be further enhanced.

[0033] In one embodiment, the peptide is an acetylated peptide such as acetyl hexapeptide-8, acetyl tetrapeptide-2, acetyl tetrapeptide-3, acetyl tetrapeptide-5, acetyl tetrapeptide-9, acetyl octapeptide-3, and acetyl oligopeptide-181; a palmitoyl series peptide such as palmitoyl tripeptide-1, palmitoyl tripeptide-5, palmitoyl pentapeptide-4, palmitoyl hexapeptide-102, and palmitoyl heptapeptide-48; and a peptide copper complex such as copper tripeptide-1 and copper palmitoyl heptapeptide-14. Polypeptides such as nonapeptide-44, nonapeptide-22, nonapeptide-35, nonapeptide-1, octapeptide-48, octapeptide-47, octapeptide-42, heptapeptide-54, and ascorbic acid polypeptide; and carnosine may be included. Specifically, at least one of acetyl hexapeptide-8, ascorbic acid polypeptide, acetyl tetrapeptide-2, acetyl tetrapeptide-3, acetyl tetrapeptide-5, acetyl tetrapeptide-9, acetyl octapeptide-3, palmitoyl tripeptide-1, palmitoyl tripeptide-5, palmitoyl pentapeptide-4, nonapeptide-1, carnosine, and copper tripeptide-1 may be included.

[0034] In the present invention, “microneedle” may have the same meaning as a spicule or a spicule. The microneedle is a composite material composed of calcium and silicate, etc., which can be obtained from freshwater sponges, and its main component is silica. The microneedle has a fine needle-like structure and can penetrate the skin layer to form micro-channels. The sponges mainly inhabit the sea in the form of sponges of various colors, such as orange or turquoise, but freshwater sponges may also exist; specifically, the sponge is *Sponzilla lacustris* ( Spongilla lacustris ), Spongilla Fragilis Lady( Spongilla fragilis Leidy ) or Epidatia pluviatilis( Ephydatia fluviatilis It can be.

[0035] In one embodiment, the microneedles may be derived from a sponge. In a specific embodiment, the microneedles may be hydrolyzed sponges.

[0037] The present invention is characterized by significantly improving the delivery of peptides to the skin by using microneedles in a form in which peptides are impregnated or coated. For example, peptides can be impregnated into the fine pores of a hydrolyzed sponge by stirring a purified hydrolyzed sponge under reduced pressure at an appropriate temperature. Since the hydrolyzed sponge penetrates the skin layer and forms micro-channels, the penetration effect of the peptides can be significantly enhanced, and it can promote skin regeneration of collagen, which acts in artificial wound healing, and increase bioactivity by raising the temperature of the skin tissue.

[0038] As such, when peptides are used in a form impregnated or coated onto microneedles, the peptides can be delivered deep into the skin along with the microneedles, thereby providing excellent skin improvement efficacy. In this case, compared to a mixture of peptides and hydrolyzed sponges, the amount of peptide delivered to the skin is significantly improved, thereby exhibiting excellent effects in reducing skin wrinkles and enhancing elasticity.

[0039] In the present invention, the term 'impregnated' of a peptide into a microneedle means that the peptide is contained on the surface of the microneedle or inside the micropores, and is a concept that includes 'coating,' and also includes a composite form in which the peptide is physically or chemically bonded to the surface of the microneedle or inside the micropores, etc.

[0040] In one embodiment, the peptide may be included in an amount of 0.01 to 0.5 weight% based on the total weight of the microneedle, specifically 0.02 to 0.3 weight%, more specifically 0.03 to 0.2 weight%, and even more specifically 0.04 to 0.1 weight%. If the peptide is included in an amount less than 0.01 weight%, the wrinkle improvement and elasticity enhancement effects may be reduced, and if it is included in an amount exceeding 0.5 weight%, the microneedle powder may clump together or uniform powder particles may not be formed.

[0041] In one embodiment, the peptide and microneedles may be included in a weight ratio of 0.01 to 0.5:99, specifically in a weight ratio of 0.03 to 0.2:99, and more specifically in a weight ratio of 0.06:99. When included in such a weight ratio, the peptide can be impregnated more uniformly into the microneedles.

[0043] In a specific embodiment, the peptide-impregnated microneedles may be provided in the form of a powder. As an example of the microneedles provided in the form said, a hydrolyzed sponge may be formed in the form of a dried powder, and it may be difficult to observe a very small and thin needle-like structure with the naked eye.

[0044] In one embodiment, the microneedles may have a length of 200 to 300 μm and a diameter of 10 to 20 μm, specifically a length of 230 to 280 μm and a diameter of 12 to 18 μm, and more specifically a length of 250 μm and a diameter of 15 μm. If the size of the microneedles is less than the above range, a large amount may penetrate into the skin and cause inflammation, and if microneedles within the above range are used, a more superior skin regeneration and self-regenerating effect may be achieved.

[0045] In one embodiment, the pH of the microneedle may be 7 to 8, specifically 7 to 7.5, and more specifically 7, but is not limited thereto.

[0047] In one embodiment, the peptide can be impregnated into the microneedle at a temperature of 45 to 55°C, specifically at 47 to 53°C, more specifically at 48 to 52°C, and even more specifically at 50°C. If the peptide is impregnated under conditions outside of the above temperature range, the microneedle powder may clump together or uniform powder particles may not be formed, and the peptide may not be uniformly impregnated into the microneedle.

[0048] In one embodiment, the peptide may be stirred for 4 to 5 hours and impregnated into the microneedles, specifically for 4.2 to 4.8 hours, more specifically for 4.5 hours and impregnated into the microneedles. If the stirring time is less than 4 hours, the peptide may not be uniformly impregnated into the microneedles, and if stirring is performed for more than 5 hours, the microneedle powder may clump together or uniform powder particles may not be formed.

[0049] In one embodiment, the peptide can be impregnated into the microneedle at a pressure of 0.04 to 0.07 MPa, specifically at 0.05 to 0.07 MPa, and more specifically at 0.06 MPa. If the pressure falls outside the above pressure range, the peptide may not be uniformly impregnated into the microneedle.

[0051] In one embodiment, peptide-impregnated microneedles may be treated with plasma before or after the peptide is impregnated. By treating the microneedles with plasma to process the surface, contaminants such as microorganisms present on the surface of the microneedles can be removed, significantly reducing the possibility of secondary contamination of the microneedles, and further enhancing the skin affinity and skin penetration of the microneedles. Accordingly, when the microneedles penetrate the skin, the compressive force at the interface formed between the microneedles and the skin is strong, thereby tightening pores present in the skin, such as pores, allowing the peptide to be delivered to the skin more effectively and preventing other foreign substances from entering through the interface. Additionally, due to the strong compressive force, surrounding cells can quickly fill the space vacated by the microneedles after they have been removed from the skin.

[0052] In one embodiment, the plasma treated on the microneedles may include atmospheric pressure plasma. 'Atmospheric pressure plasma' refers to plasma capable of discharge in open air without separate vacuum treatment; in addition to the intrinsic excitation species of the discharge gas, it can react with oxygen, water, or nitrogen to generate various reactive oxygen species or chemical species with metastable excited state energy. In particular, atmospheric pressure plasma may have very high physical / chemical reactivity because high-temperature electrons and low-temperature ion particles coexist in a state of non-thermodynamic equilibrium. In specific embodiments, the atmospheric pressure plasma may be treated by a method selected from the group consisting of corona discharge, dielectric barrier discharge (DBD), atmospheric pressure glow discharge, and jet methods. In particular, when treated by the jet method, contaminants on the microneedles are effectively removed, and the affinity and penetration power for the skin may be even superior.

[0053] In one embodiment, the atmospheric pressure plasma may be an atmospheric pressure low-temperature plasma. Low-temperature plasma refers to a plasma at a temperature in which electrons and ions constituting the plasma are in a state of thermodynamic disequilibrium and can have high reactivity. In a specific embodiment, the temperature of the atmospheric pressure low-temperature plasma bulk may be 1000K or lower, and the temperature of the peripheral part of the bulk may be 10 to 90℃.

[0054] In one embodiment, atmospheric pressure plasma can be generated from a low-frequency input frequency of 100 kHz or less. When using the low-frequency input frequency, low-power operation is possible, and the miniaturization of the discharge device and power supply device is very easy, so the process can be carried out economically. However, it is not limited thereto, and any input frequency capable of generating atmospheric pressure plasma can be used without limitation. As examples of the input frequency, RF plasma using an RF (Radio Frequency) generator can use a frequency of 13.56 MHz or 27.12 MHz, and MW (Microwave) plasma can use a frequency of 2.45 GHz.

[0055] In one embodiment, the atmospheric pressure plasma may be generated using one or more gases selected from the group consisting of Clean Dry Air, carbon dioxide, nitrogen, oxygen, argon, neon, and helium. Specifically, oxygen or argon gas may be used, and when oxygen or argon gas is used, a superior microneedle surface may be formed.

[0056] In one embodiment, the gas generating the atmospheric pressure plasma can be supplied at a pressure of 0.07 to 0.6 MPa, specifically at 0.07 to 0.6 MPa, and more specifically at 0.4 to 0.6 MPa. If the gas is supplied at less than 0.07 MPa, the pressure may be too low to perform uniform surface processing, and if it is supplied at more than 0.6 MPa, the pressure may be too high to control the gas flow or unstable plasma may be generated.

[0058] According to another embodiment of the present invention, a cosmetic composition comprising microneedles impregnated with peptide may be provided.

[0059] In one embodiment, a cosmetic composition comprising peptide-impregnated microneedles may have an excellent wrinkle-improving effect.

[0060] In addition, the cosmetic composition according to the present invention may be prepared in one or more formulations selected from the group consisting of skin lotion, skin softener, skin toner, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, and body cleanser.

[0061] In addition, the cosmetic composition according to the present invention may further include ingredients included in general cosmetic compositions, such as, for example, functional ingredients, purified water, preservatives, moisturizers, electrolyte compounds, ion-chelating agents, disinfectants, pH adjusters, antioxidants, alcohols, fragrances, etc.

[0062] The amounts of the aforementioned ingredients are not particularly limited and can be easily selected by a person skilled in the art within a range that does not impair the purpose and effects of the present invention.

[0064] In the following, examples and experimental examples are presented to further explain the present invention in more detail, but the present invention is not limited thereto.

[0066] Preparation Example

[0067] (1) Preparation of Example 1

[0069] Example 1 was prepared by following steps 1) to 5).

[0071] 1) Microneedle purification

[0072] Raw freshwater sponges inhabiting fresh water were purchased and stirred for 30 minutes while raising the temperature to 40°C. After reaching 40°C, hydrolysis was carried out while stirring for an additional 48 hours.

[0074] 2) Primary sieving and removal of foreign substances

[0075] The raw material hydrolyzed in 1) above was sieved through 200 mesh, and the hydrolyzed sponge (silica) was separated and washed 5 times. The hydrolyzed sponge was placed in a Nuche filter and removed by repeating the process 5 times, then the solid impurities were removed by repeating the centrifugation process 3 times, and the impurities were removed by repeating the sedimentation extraction process 3 times.

[0077] 3) Secondary sieving after drying

[0078] The hydrolyzed sponge obtained in 2) above was dried at 80°C for 24 hours, and then sieved once again with 200 mesh.

[0080] 4) Peptide impregnation and coating

[0081] After reducing the pressure of the vacuum stirrer to 50℃ and 0.06MPa, the hydrolyzed sponge obtained in 3) above and the peptide solution (peptide concentration: 5.6wt%, solvent: water) were simultaneously added in a weight ratio of 99:1 and stirred for 4 hours and 30 minutes to impregnate and coat the skin active ingredients into the hydrolyzed sponge. Specifically, the peptide solution used 13 types of peptide solutions including acetyl hexapeptide-8, ascorbic acid polypeptide, acetyl tetrapeptide-2, acetyl tetrapeptide-3, acetyl tetrapeptide-5, acetyl tetrapeptide-9, acetyl octapeptide-3, palmitoyl tripeptide-1, palmitoyl tripeptide-5, palmitoyl pentapeptide-4, nonapeptide-1, carnosine, and copper tripeptide-1, and the 13 types of peptides are contained inside liposomes.

[0083] 5) Plasma treatment

[0084] Plasma was applied to a hydrolyzed sponge impregnated and coated with the peptide obtained through 4) above. Specifically, an atmospheric pressure low-temperature plasma having a plasma output of 1000W, a discharge voltage of 8 to 12kV, and a discharge frequency of 40 to 50kHz was generated using an input power of 220V, an input frequency of 60Hz, Clean Dry Air as the gas, a gas pressure of 0.5MPa, and a gas flow rate of 50L / min, and was applied to the hydrolyzed sponge.

[0086] (2) Confirmation of microneedles impregnated with active ingredients

[0088] To confirm whether the hydrolyzed sponge of Example 1 was impregnated with an active ingredient, the microneedles of Example 1 were examined using a scanning electron microscope (SEM; S-4800 from HITACHI). As a result, it was confirmed that the peptide was well impregnated into the microneedles. Figure 1a shows a magnified photograph of Example 1 taken under a microscope, and Figure 1b shows a magnified photograph of the microneedles before peptide impregnation.

[0090] Experimental Example 1. Comparative experiment based on the presence or absence of plasma treatment

[0092] In order to compare the effects depending on whether or not the plasma process treatment is performed in step 5) of the above manufacturing process, Example 2 was prepared by excluding the plasma treatment process in step 5) of the above manufacturing example and going through steps 1) to 4).

[0094] (1) Evaluation of skin absorption rate through Franz infusion cell assay

[0096] The peptide delivery amount was measured for the above Examples 1 and 2. The specific measurement method is as follows.

[0097] The skin penetration device consists of a donor chamber and a receptor chamber, and the prepared skin was fixed to the device between them with the stratum corneum facing upward. Figure 2 shows a schematic diagram of the skin penetration device used in this experiment. The prepared skin is dorsal or thoracic skin with a thickness of 200–400 μm collected using a skin collection device. Physical damage was evaluated via visual inspection, and unsuitable skin was removed. Integrity was ensured by verifying that the alternating electrical resistance was within the normal range for skin (2 volts or higher). A glass skin penetration device was used to minimize reaction with the experimental substance. The flow-through diffusion method was used for the sample. Since physiologically inactive skin was used, 6% polyethylene glycol 20-oleyl ether solution or 5% bovine serum albumin was added to physiological saline solution at pH 7.4.

[0098] Experiments were conducted for Examples 1 and 2. In addition, to reference the skin penetration performance of general peptides, the peptide solution of the above preparation example (peptide concentration: 5.6 wt%, solvent: water) was used as a control to verify skin penetration. Each substance was quantitatively analyzed using a 1260 infinity 2 HPLC (Agilent, USA) by searching the literature for suitable analysis conditions and establishing the conditions, and the transmittance was calculated using this.

[0099] The samples were prepared in liquid form and placed in the donor compartment. In experiments using hydrolyzed sponges, simply placing them on the skin was not sufficient to affect the permeability; therefore, a small amount of water was added to Examples 1 and 2 to create a liquid, and 0.5g was slowly applied to fixed skin using a spatula. For all experiments, samples were taken after a set period of time, and the permeated substances were analyzed. The results of the above experiments are shown in Table 1 below.

[0100]

[0102] As shown in Table 1 above, when the amount absorbed into the skin of the control group was set to 1, the absorption rate of Example 1, in which plasma was applied to microneedles impregnated with peptides, increased up to 4.3 times, showing a significantly superior absorption rate, and Example 2, in which peptides were impregnated and not treated with plasma, showed a superior absorption rate of 2.5 times.

[0104] (2) Evaluation of skin improvement efficacy

[0106] To confirm the skin improvement effects of Examples 1 and 2 above, the effectiveness of skin elasticity enhancement, wrinkle improvement, and moisturizing was evaluated on 20 men and women in their 50s who volunteered to report decreased skin elasticity and increased wrinkles. Specifically, the 20 men and women were divided into two groups, A and B, with 10 people in each group. Example 1 was applied to Group A and Example 2 to Group B every morning after washing their faces, and the skin improvement effects were evaluated after 4 weeks. Specifically, each subject was asked to take and compare photographs of their face on the start and end days of the experiment. After comparing the two photos, the subjects were asked to rate on a scale of 1 to 9 (where a score closer to 9 indicates superior efficacy) whether they felt their skin had elasticity, wrinkles had improved, and that sufficient moisture was maintained on the skin for a long period after application. Subsequently, the scores of the 20 subjects were aggregated to confirm the effects of skin elasticity, wrinkle improvement, and moisturizing. The results of the experiment are shown in Table 2 below.

[0107]

[0109] As shown in Table 2 above, regarding elasticity enhancement and wrinkle improvement, Examples 1 and 2, in which peptides were impregnated into microneedles, both received scores approaching or exceeding an average of 7 points, confirming that they have excellent efficacy in enhancing skin elasticity and improving wrinkles. In particular, Example 1, in which plasma was applied to the microneedles, received an even higher score, indicating very excellent efficacy in enhancing skin elasticity and improving wrinkles. Furthermore, regarding moisturizing efficacy, Examples 1 and 2, in which peptides were impregnated into microneedles, both received high scores exceeding an average of 7 points, confirming that they have excellent skin moisturizing efficacy. In particular, Example 1, in which plasma was applied to the microneedles, received an even higher score, indicating very excellent skin moisturizing efficacy.

[0111] Experimental Example 2. Comparative experiment based on the presence or absence of peptide coating

[0112] To compare the effects of a hydrolyzed sponge with or without a peptide coating, 4) Comparative Example 1 was prepared by simply mixing (hand-mixing) the hydrolyzed sponge that had undergone steps 1) to 3) below and the peptide solution, excluding the coating process.

[0114] (1) Evaluation of skin absorption rate through Franz infusion cell assay

[0116] The peptide delivery amount was measured for Comparative Example 1 prepared above. The specific measurement method is the same as in Experimental Example 1. The experimental results are shown in Table 3 below.

[0117]

[0119] As shown in Table 3 above, Comparative Example 1, which simply mixed microneedles and peptides, showed an absorption rate approximately 1.2 times higher than the control group, but was measured to have a significantly reduced absorption rate compared to the examples in which peptides were directly impregnated into the microneedles. Therefore, it was confirmed that the skin absorption rate of peptides significantly increased in hydrolyzed sponges impregnated or coated with peptides compared to cases where microneedles and peptides were simply mixed.

[0121] (2) Evaluation of skin improvement efficacy

[0123] Skin elasticity, wrinkle improvement, and moisturizing effects were evaluated for Comparative Example 1 using the same method as in Experimental Example 1 above. The experimental results are shown in Table 4 below.

[0124]

[0126] As shown in Table 4 above, regarding skin elasticity enhancement and wrinkle improvement, Comparative Example 1, in which peptides were not impregnated into the microneedles, showed a significantly lower average score compared to the Example, indicating that the skin elasticity enhancement and wrinkle improvement effects were inferior. In addition, regarding skin moisturizing efficacy, Comparative Example 1, in which peptides were not impregnated into the microneedles, showed a significantly lower average score compared to the Example, indicating that the skin moisturizing effect was inferior. Therefore, it was evaluated that excellent skin elasticity enhancement, wrinkle improvement, and skin moisturizing effects were exhibited as the amount of delivery to the skin was enhanced by impregnating the microneedles with peptides.

[0128] Experimental Example 3. Comparative experiment according to impregnation conditions

[0130] In order to confirm the conditions for forming microneedles impregnated with peptides of excellent physical properties, comparative examples were prepared by changing the process conditions of “4) peptide impregnation and coating” in the above preparation example (1). Specifically, as shown in Table 5, comparative examples 2 and 3 were prepared by varying the conditions of the amplification stirrer.

[0131]

[0133] (1) Check whether excellent powder particles are formed

[0135] It was confirmed whether the comparative example prepared according to the above process conditions formed a uniform powder with a uniform particle size. As a result of preparation, Example 1, impregnated under appropriate temperature and stirring time, formed uniform particles evenly. On the other hand, Comparative Example 2, impregnated under conditions exceeding the appropriate temperature and less than the appropriate stirring time, and Comparative Example 3, impregnated under conditions lower than the appropriate temperature and more than the appropriate stirring time, formed non-uniform particles, and a phenomenon of particles clumping together was observed. Figure 3 shows the microneedles, Example 1, and Comparative Example 2 before the powder impregnation process.

[0137] (2) Evaluation of skin improvement efficacy

[0139] Skin elasticity, wrinkle improvement, and moisturizing effects were evaluated for Comparative Examples 2 and 3 using the same method as in Experimental Example 1 above. The experimental results are shown in Table 6 below.

[0140]

[0142] As shown in Table 6 above, Comparative Examples 2 and 3, which were impregnated under conditions deviating from the appropriate process temperature and stirring time, showed superior skin improvement efficacy compared to Comparative Example 1, which was not impregnated with peptide; however, they were given significantly lower evaluation scores compared to Example 1, in which uniform and even powder particles were formed. Therefore, it was evaluated that the peptide was not properly impregnated or coated onto the microneedles when the appropriate process temperature and stirring time were deviated from. Thus, it was confirmed that impregnating or coating the peptide onto the microneedles through appropriate temperature and stirring time is an important factor in obtaining microneedles with excellent physical properties.

[0144] Foregoing, specific parts of the content of the present invention have been described in detail. It will be apparent to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the present invention. Accordingly, the actual scope of the present invention is defined by the appended claims and their equivalents.

Claims

Claim 1 A peptide-impregnated microneedle, wherein the microneedle comprises a hydrolyzed sponge, the peptide is impregnated and coated onto the microneedle by stirring at 45 to 55°C for 4 to 5 hours, and plasma is treated after the peptide is impregnated and coated onto the microneedle. Claim 2 delete Claim 3 A microneedle according to claim 1, wherein the peptide comprises at least one of an acetylated peptide; a palmitoyl-based peptide; a peptide copper complex; a polypeptide; and carnosine. Claim 4 In claim 1, the peptide comprises at least one of acetyl hexapeptide-8, ascorbic acid polypeptide, acetyl tetrapeptide-2, acetyl tetrapeptide-3, acetyl tetrapeptide-5, acetyl tetrapeptide-9, acetyl octapeptide-3, palmitoyl tripeptide-1, palmitoyl tripeptide-5, palmitoyl pentapeptide-4, nonapeptide-1, carnosine, and copper tripeptide-1, forming a microneedle. Claim 5 In claim 1, the microneedle comprises the peptide contained inside a liposome. Claim 6 delete Claim 7 delete Claim 8 In claim 1, the peptide is impregnated into the microneedle at 0.04 to 0.07 MPa. Claim 9 In claim 1, the microneedle is included in an amount of 0.01 to 0.5 weight% based on the total weight of the microneedle. Claim 10 delete Claim 11 A microneedle according to claim 1, characterized in that the plasma is an atmospheric pressure low-temperature plasma. Claim 12 A cosmetic composition having a wrinkle-improving effect, comprising a microneedle according to any one of claims 1, 3 to 5, 8, 9 and 11. Claim 13 delete

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