Animal placenta protein anti-aging, wrinkle-fading, brightening and skin-care formula

Abstract

The invention relates to an anti-aging, wrinkle-fading, brightening and skin-care formula and a preparation method. The formula comprises a core active component, an auxiliary active component, a moisturizing additive, a soothing and repairing agent, a preservative, a water-phase basic auxiliary material and deionized water, the core active component contains modified sheep placenta small molecular protein, acetyl hexapeptide-8 and glutathione; the preparation method comprises the steps of water-phase matrix preparation, primary mixing, active component adding and dispersing, pH adjusting, curing and the like, and sheep placenta micromolecular protein is subjected to pretreatment, enzymolysis and performance modification and optimization. The formula components are scientifically compounded, the preparation process is accurate, and the product stability and mildness are excellent; the composition can effectively increase the collagen content of the skin, reduce fine wrinkles and brighten the skin color, has good moisturizing and preservative effects, and has a use effect superior to that of a conventional skin care formula.

Description

Technical Field

[0001] This invention relates to the field of skincare technology, specifically to an anti-aging, wrinkle-reducing, and brightening skincare formula using animal placental protein. Background Technology

[0002] With the development of the skincare industry, anti-aging, wrinkle reduction, and brightening have become core skincare needs for the general public. Animal placental protein, rich in amino acids, growth factors, and other active substances, can effectively activate skin collagen regeneration, making it an important raw material in the anti-aging skincare field. However, existing skincare formulas containing animal placental protein still have many technical shortcomings in practical applications, which restrict their efficacy and product experience.

[0003] Current technologies for processing sheep placental protein often employ single-protease hydrolysis. This not only results in incomplete degreasing and deodorization pretreatment, leaving residual odors that affect product usability, but also leads to uneven molecular weight distribution of the hydrolysate. Large protein molecules are difficult to absorb through the skin, while small active fragments are easily lost. Although some technologies attempt to modify placental protein, the modification methods are singular, and the degree of phosphorylation modification is poorly controlled, resulting in low protein bioavailability and difficulty in fully realizing anti-aging activity. Furthermore, the formulation of active ingredients in existing products lacks scientific rigor. Anti-aging, wrinkle-reducing, and brightening components are often simply mixed without achieving targeted synergistic effects. For example, unmodified ceramides (NPs) have poor stability and are prone to compatibility issues with collagen components, leading to easy degradation of their activity. Meanwhile, the ratio of brightening components such as niacinamide and ascorbate glucoside to placental protein is unreasonable, easily resulting in antagonistic effects and making it difficult to simultaneously achieve the triple effects of anti-aging, wrinkle reduction, and brightening. In addition, the existing formulas have flawed system design. The moisturizing aids are mostly single ingredients, with short-lived moisturizing effects and poor compatibility with active ingredients. The preservative system often contains alcohols or parabens, which are highly irritating and can easily cause discomfort to sensitive skin. The soothing and repairing ingredients are of a single type and cannot alleviate the slight irritation of the active ingredients to the skin. Overall, the formula is not gentle enough and has poor compatibility.

[0004] In terms of preparation processes, existing methods often lack precise temperature and rotation speed control, and the addition of active ingredients is often crude. This makes them prone to inactivation due to excessive high temperatures or shear forces. Furthermore, the lack of fine dispersion of the mixture results in uneven particle size distribution and poor transdermal absorption, ultimately leading to a significant reduction in the stability and actual skincare efficacy of the final product. In summary, existing skincare formulas containing sheep placental protein struggle to balance efficacy, gentleness, stability, and transdermal absorption. There is an urgent need to develop a modified, optimized, synergistically formulated, and precisely processed sheep placental protein anti-aging, wrinkle-reducing, and brightening skincare formula to address the shortcomings of existing technologies. Summary of the Invention

[0005] (a) Technical problems to be solved To address the shortcomings of existing technologies, this invention provides an anti-aging, wrinkle-reducing, and brightening skincare formula using animal placental protein.

[0006] (II) Technical Solution A skincare formula for anti-aging, wrinkle reduction, and brightening using animal placental protein, comprising, by weight, a combination of core active ingredients and auxiliary active ingredients. The core active ingredients are: 1.5-2.5 parts of sheep placental small molecule protein, 0.08-0.15 parts of acetyl hexapeptide-8, and 0.04-0.08 parts of glutathione. The sheep placental small molecule protein undergoes defatting and deodorization pretreatment, stepwise enzymatic hydrolysis by a ternary complex protease, and phosphorylation modification. The modified sheep placental small molecule protein has a molecular weight of 1500-3500 Da and a phosphorylation modification degree of 30%-50%. The auxiliary active ingredients are: 3.0-4.0 parts of ascorbate glucoside, 0.8-1.2 parts of hydrogenated ceramide NP, 4.0-5.0 parts of niacinamide, 2.0-3.0 parts of hydrolyzed fish collagen, 0.15-0.25 parts of sodium hyaluronate, and 1.5-2.0 parts of panthenol.

[0007] Preferably, the product further comprises 8.0-12.0 parts by weight of a compound moisturizing agent, wherein the compound moisturizing agent is a mixture of butanediol, 1,3-propanediol, and polyoxyethylene ether modified glycerin in a mass ratio of 3:2:1.

[0008] Preferably, the ternary complex protease is composed of papain, neutral protease, and bromelain, and the enzyme activity ratio is 2:1:1.

[0009] Preferably, the product further comprises 0.1-0.2 parts by weight of a highly active soothing and repairing agent, wherein the soothing and repairing agent is a compound of dipotassium glycyrrhizate, allantoin, and zinc pyrrolidone carboxylate, a panthenol derivative, in a mass ratio of 2:1:1, wherein the dipotassium glycyrrhizate has an activity content of ≥98% and the allantoin has a purity of ≥99%.

[0010] Preferably, the product further includes 0.2-0.4 parts by weight of an alcohol-free compound preservative, wherein the preservative is a compound of p-hydroxyacetophenone, capryloyl hydroxamic acid, 1,2-hexanediol, and ethylhexylglycerin in a mass ratio of 3:2:2:1. The antibacterial spectrum of the preservative system covers Gram-positive bacteria, Gram-negative bacteria, and molds, with a minimum inhibitory concentration of 0.05%-0.1%.

[0011] Preferably, the stepwise enzymatic hydrolysis process parameters of the ternary complex protease are as follows: first, the protease is enzymatically hydrolyzed for 2 hours at 50-55℃ and pH 6.0-6.5; then, the protease is further hydrolyzed for 1 hour at 45-50℃ and pH 7.0-7.5 by a mixture of neutral protease and bromelain; after hydrolysis, the enzyme is inactivated at 100℃ for 10 minutes; and after centrifugation and filtration, a sheep placental protein solution with a molecular weight of 1500-3500 Da is obtained. Sodium pyrophosphate is used as the phosphorylation agent for phosphorylation modification at 40℃ for 3 hours.

[0012] Preferably, the preparation method of the animal placental protein anti-aging, wrinkle-reducing, and brightening skin care formula includes the following steps: S1. Weigh the amount of aqueous basic excipients in the formula and add them to ultrapure deionized water with a conductivity of ≤10μS / cm. Heat the water in a water bath at 82-84℃ and simultaneously homogenize and shear at 3500-3800rpm for 12-14min to obtain a homogeneous aqueous matrix. S2. Cool the aqueous matrix to 57-59℃ at a rate of 2-3℃ / min, add the formulated amount of compound moisturizing agent, ascorbate glucoside and nicotinamide, and stir at 800-1000 rpm for 11-12 min to dissolve and obtain a primary mixture without precipitate. S3. Cool the primary mixture to 42-44℃ at a rate of 1-2℃ / min. First, add the modified sheep placenta small molecule protein and hydrolyzed fish collagen according to the formula and stir for 5 minutes. Then, add acetyl hexapeptide-8, glutathione, and hydrogenated modified ceramide NP in stepwise order. Finally, add sodium hyaluronate and panthenol. Stir continuously at 1200-1500 rpm throughout the process to obtain the core active mixture. S4. Add the formulated amount of highly active soothing and repairing agent and alcohol-free compound preservative to the core active mixture. Adjust the pH of the system to 5.8-6.2 using a mixture of citric acid and arginine. Stir at 600-700 rpm during the adjustment process. After adjustment, continue stirring for 6-7 minutes. Filter through a filter membrane to obtain the finished skin care formula.

[0013] Preferably, the aqueous matrix prepared in S1 needs to be immediately subjected to vacuum degassing treatment, with a degassing pressure of -0.08 to -0.1 MPa and a degassing time of 10 min. The solid content of the aqueous matrix after degassing is 5%-8%, and the shearing force of the homogeneous shearing is controlled at 20-25 MPa.

[0014] Preferably, the peptides, proteins and ceramide components in S3 are added in steps at intervals of 1.5 min. The citric acid and arginine mixture in S4 is compounded at a mass ratio of 1:2. The pH is adjusted using a gradient adjustment method, first coarsely adjusted to 7.0-7.5 and then finely adjusted to 5.8-6.2. The filter membrane is a 200-mesh nylon filter membrane. After filtration, the finished product is allowed to stand and mature for 24 h at 25°C in the dark.

[0015] Preferably, after all the core active components in S3 are added, the core active mixture is subjected to low-frequency ultrasonic dispersion treatment. The ultrasonic frequency is 20-30kHz, the ultrasonic power is 100-150W, and the ultrasonic time is 3-5min. After ultrasonic treatment, the particle size distribution of the core active mixture is 100-200nm, and the temperature of the mixture is maintained at 42-44℃ during the ultrasonic process.

[0016] (iii) Beneficial technical effects Compared with existing technologies, the beneficial effects of this invention are: The core sheep placental protein undergoes degreasing and deodorization pretreatment, stepwise enzymatic hydrolysis with a ternary complex protease, and phosphorylation modification, resolving the issues of residual odor and uneven molecular weight in traditional placental proteins. The modified protein retains its full activity, significantly improving transdermal absorption and allowing it to better penetrate deep into the skin, fully exerting its anti-aging activity. Furthermore, it is non-allergenic, enhancing the safety of the formula. The scientifically designed ratio of core and auxiliary active components in the formula ensures targeted synergistic effects from various anti-aging, wrinkle-reducing, and brightening components, effectively avoiding efficacy antagonism. This results in a synergistic enhancement of the triple skincare effects of anti-aging, wrinkle reduction, and brightening. Compared to existing simple compound formulas, the overall skincare effect is superior, addressing the root causes of skin aging, fine lines, and dullness.

[0017] The optimized design of the formulation system further enhances the overall performance of the product. The compound moisturizing aid provides long-lasting hydration and works synergistically with sodium hyaluronate to maintain skin hydration for an extended period. The alcohol-free compound preservative has good antibacterial effects and is gentle and non-irritating, eliminating the need for traditional irritating preservatives. The highly active soothing and repairing agents enhance the repairing ability and effectively relieve skin discomfort, making the formula suitable for various skin types, including sensitive skin, and thus applicable to a wider range of skin types. The preparation process of this invention employs refined operations such as gradient cooling, stepwise addition, vacuum degassing, and low-frequency ultrasonic dispersion, which can precisely control various process parameters, effectively preventing the inactivation of active ingredients due to high temperature and excessive shear force. At the same time, it makes the particle size distribution of the finished product more uniform, further improving the transdermal absorption effect and making the active ingredients easier for the skin to absorb and utilize. Attached Figure Description

[0018] Figure 1 This is a flowchart of the preparation method of the animal placental protein anti-aging, wrinkle-reducing, and brightening skin care formula disclosed in this invention; Figure 2 This is a line graph comparing the zeta potential and transdermal absorption rate of the core active ingredient in the examples and comparative examples; Figure 3 This is a bar chart comparing the skin collagen content increase rate after 28 days and the fine line depth reduction rate after 42 days between the examples and the comparative examples; Figure 4 This is a radar comparison chart created by standardizing the dimensions of the performance comparison data of the examples and comparative examples. Detailed Implementation

[0019] according to Figures 1 to 4 The specific embodiments of the present invention are as follows: The following detailed description of the animal placental protein anti-aging, wrinkle-reducing, and brightening skincare formula and preparation method of the present invention, in conjunction with specific embodiments and comparative examples, is provided. Components not explicitly mentioned are all ultrapure deionized water with a conductivity ≤10μS / cm. All performance tests were conducted in accordance with skincare industry standards and conventional testing methods.

[0020] I. Raw Material Preparation (I) Core active ingredient raw materials 1. Raw materials for preparing small molecule protein from sheep placenta: fresh sheep placenta, papain (enzyme activity ≥8000U / g), neutral protease (enzyme activity ≥6000U / g), bromelain (enzyme activity ≥7000U / g), sodium pyrophosphate (food grade), ethanol (analytical grade), activated carbon (food grade).

[0021] 2. Other core active components: Acetyl hexapeptide-8, glutathione.

[0022] (ii) Auxiliary active ingredient raw materials Ascorbate glucoside, hydrogenated ceramide NP, nicotinamide, hydrolyzed fish collagen with a molecular weight ≤5000 Da, sodium hyaluronate with a molecular weight of 100-200 kDa, and panthenol.

[0023] (iii) Other auxiliary materials 1. Compound moisturizing aids: Butylene glycol (food grade), 1,3-propanediol (food grade), polyoxyethylene ether modified glycerin (hydroxyl value 300-350 mg KOH / g), compounded in a mass ratio of 3:2:1.

[0024] 2. Highly active soothing and repairing agents: dipotassium glycyrrhizate, allantoin, and zinc pyrrolidone carboxylate, compounded in a mass ratio of 2:1:1.

[0025] 3. Alcohol-free compound preservative: p-hydroxyacetophenone, capryloyl hydroxamic acid, 1,2-hexanediol, and ethylhexylglycerin are compounded in a mass ratio of 3:2:2:1.

[0026] 4. pH adjuster: Citric acid and arginine, mixed in a mass ratio of 1:2.

[0027] 5. Aqueous phase base materials: Xanthan gum, food grade, viscosity ≥10000mPa·s, carbomer.

[0028] II. Example 1 (a) Formula composition (by mass parts) 1. Core active ingredients: 1.5 parts of sheep placental small molecule protein, 0.08 parts of acetyl hexapeptide-8, and 0.04 parts of glutathione.

[0029] 2. Auxiliary active components: 3.0 parts of ascorbate glucoside, 0.8 parts of hydrogenated modified ceramide NP, 4.0 parts of nicotinamide, 2.0 parts of hydrolyzed fish collagen, 0.15 parts of sodium hyaluronate, and 1.5 parts of panthenol.

[0030] 3. Compound moisturizing agent: 8.0 parts.

[0031] 4. Highly active soothing and repairing agent: 0.1 parts.

[0032] 5. Alcohol-free compound preservative: 0.2 parts.

[0033] 6. Aqueous phase base materials: 0.1 parts xanthan gum, 0.08 parts carbomer.

[0034] (II) Preparation of small molecule proteins from sheep placenta 1. Degreasing and deodorizing pretreatment: Take fresh sheep placenta, rinse it 3 times with ultrapure deionized water, remove impurities such as fascia and blood vessels, and cut it into 1cm pieces. 3 Small pieces of placenta were placed in a high-speed homogenizer, and 3 times the mass of ultrapure deionized water were added. The homogenizer was homogenized at 10,000 rpm for 20 minutes to obtain a sheep placenta homogenate. 2 times the volume of ethanol was added to the homogenate, and the mixture was refluxed in a 50°C water bath for 1 hour. The precipitate was collected by centrifugation at 8,000 rpm for 20 minutes to remove fat. 0.5% activated carbon was added to the precipitate, and the mixture was stirred and adsorbed at 40°C for 30 minutes. The activated carbon was removed by filtration to obtain a degreased and deodorized sheep placenta tissue fluid.

[0035] 2. Stepwise enzymatic hydrolysis using a ternary complex protease: Papain, neutral protease, and bromelain were weighed at an enzyme activity ratio of 2:1:1, with the total enzyme addition amount being 1.5% of the sheep placental tissue fluid mass. First, the pH of the sheep placental tissue fluid was adjusted to 6.0, and the temperature was raised to 50℃. Papain was added, and the mixture was incubated for 2 hours, with stirring for 10 minutes every 30 minutes at a speed of 500 rpm. Then, the pH of the system was adjusted to 7.0, and the temperature was lowered to 45℃. A mixture of neutral protease and bromelain was added, and the enzymatic hydrolysis continued for 1 hour, with the stirring method as before. After the enzymatic hydrolysis was completed, the system was heated to 100℃ and incubated for 10 minutes to inactivate the enzyme. Then, it was cooled to room temperature, centrifuged at 8000 rpm for 20 minutes, and the supernatant was filtered through a 100-mesh filter to obtain sheep placental protein solution with a molecular weight of 1500-3500 Da.

[0036] 3. Phosphorylation modification: 5% sodium pyrophosphate was added to the sheep placental protein solution as a modifier. The mixture was stirred at 40°C for 3 hours at a speed of 300 rpm. After modification, the unreacted sodium pyrophosphate was removed by dialysis with a molecular weight cutoff of 1000 Da for 24 hours. After freeze-drying, the modified sheep placental small molecule protein was obtained. The degree of phosphorylation modification was found to be 30%.

[0037] (III) Skin Care Formula Preparation S1: Weigh the xanthan gum and carbomer according to the formula, add them to 78.45 parts of ultrapure deionized water, place them in an 82℃ constant temperature water bath, and start the homogenizing shear machine at 3500 rpm and 20 MPa shear force for 12 min to obtain a homogeneous aqueous matrix; immediately transfer the aqueous matrix to a vacuum degassing machine and degas at -0.08 MPa pressure for 10 min. After degassing, the solid content of the aqueous matrix is ​​measured to be 5%.

[0038] S2: Cool the degassed aqueous matrix to 57°C at a rate of 2°C / min, add the formulated amount of compound moisturizing agent, ascorbate glucoside, and nicotinamide, start the stirrer and stir at 800 rpm for 11 min to dissolve, observe every 2 min to ensure no precipitation occurs, and obtain the primary mixture.

[0039] S3: Cool the primary mixture to 42°C at a rate of 1°C / min. First, add the prepared modified sheep placental small molecule protein and hydrolyzed fish collagen, and stir at 800 rpm for 5 min until completely dispersed. Then, add acetyl hexapeptide-8, glutathione, and hydrogenated modified ceramide NP in stepwise order, with an interval of 1.5 min between each two components. Finally, add sodium hyaluronate and panthenol, and continuously stir at 1200 rpm throughout the process. After all the core active components have been added, transfer the mixture to a low-frequency ultrasonic disperser, set the ultrasonic frequency to 20 kHz and the ultrasonic power to 100 W, and ultrasonically treat for 3 min. During the ultrasonic treatment, maintain the temperature of the mixture at 42°C using a constant temperature water bath. The particle size distribution of the core active mixture after ultrasonication is 100-150 nm, as measured by a Malvern laser particle size analyzer.

[0040] S4: Add the formulated amount of highly active soothing and repairing agent and alcohol-free compound preservative to the core active mixture. Adjust the pH of the system using a mixture of citric acid and arginine, first coarsely adjusting to 7.0, then finely adjusting to 5.8. Stir at 600 rpm during the adjustment process, and continue stirring for 6 minutes after adjustment. Filter the mixture through a 200-mesh nylon filter membrane at a filtration pressure of 0.3 MPa, collect the filtrate, and let it stand and mature at 25°C in the dark for 24 hours to obtain the finished skin care formula.

[0041] III. Example 2 (a) Formula composition (by mass parts) 1. Core active ingredients: 2.0 parts of sheep placental small molecule protein, 0.12 parts of acetyl hexapeptide-8, and 0.06 parts of glutathione.

[0042] 2. Auxiliary active components: 3.5 parts ascorbate glucoside, 1.0 part hydrogenated ceramide NP, 4.5 parts nicotinamide, 2.5 parts hydrolyzed fish collagen, 0.20 parts sodium hyaluronate, and 1.7 parts panthenol.

[0043] 3. Compound moisturizing agent: 10.0 parts.

[0044] 4. Highly active soothing and repairing agent: 0.15 parts.

[0045] 5. Alcohol-free compound preservative: 0.3 parts.

[0046] 6. Aqueous phase base materials: 0.12 parts xanthan gum, 0.10 parts carbomer.

[0047] (II) Preparation of small molecule proteins from sheep placenta 1. Degreasing and deodorizing pretreatment: Take fresh sheep placenta, rinse it 3 times with ultrapure deionized water, remove impurities such as fascia and blood vessels, and cut it into 1cm pieces. 3 Small pieces of placenta were placed in a high-speed homogenizer, and 3 times the mass of ultrapure deionized water were added. The homogenizer was homogenized at 10,000 rpm for 20 minutes to obtain a sheep placenta homogenate. 2 times the volume of ethanol was added to the homogenate, and the mixture was refluxed in a 50°C water bath for 1 hour. The precipitate was collected by centrifugation at 8,000 rpm for 20 minutes to remove fat. 0.5% activated carbon was added to the precipitate, and the mixture was stirred and adsorbed at 40°C for 30 minutes. The activated carbon was removed by filtration to obtain a degreased and deodorized sheep placenta tissue fluid.

[0048] 2. Stepwise enzymatic hydrolysis using a ternary complex protease: Papain, neutral protease, and bromelain were weighed at an enzyme activity ratio of 2:1:1, with the total enzyme addition amount being 1.5% of the sheep placental tissue fluid mass. First, the pH of the sheep placental tissue fluid was adjusted to 6.3, and the temperature was raised to 53℃. Papain was added, and the mixture was incubated for 2 hours, with stirring for 10 minutes every 30 minutes at a speed of 500 rpm. Then, the pH of the system was adjusted to 7.3, and the temperature was lowered to 48℃. A mixture of neutral protease and bromelain was added, and the enzymatic hydrolysis continued for 1 hour, with stirring as before. After the enzymatic hydrolysis was completed, the system was heated to 100℃ and incubated for 10 minutes to inactivate the enzyme. Then, it was cooled to room temperature, centrifuged at 8000 rpm for 20 minutes, and the supernatant was filtered through a 100-mesh filter to obtain sheep placental protein solution with a molecular weight of 1500-3500 Da.

[0049] 3. Phosphorylation modification: 5% sodium pyrophosphate was added to the sheep placental protein solution as a modifier. The mixture was stirred at 40°C for 3 hours at a speed of 300 rpm. After modification, the unreacted sodium pyrophosphate was removed by dialysis with a molecular weight cutoff of 1000 Da for 24 hours. After freeze-drying, the modified sheep placental small molecule protein was obtained. The degree of phosphorylation modification was found to be 40%.

[0050] (III) Skin Care Formula Preparation S1: Weigh the xanthan gum and carbomer according to the formula, add them to 73.75 parts of ultrapure deionized water, place them in an 83℃ constant temperature water bath, and homogenize them at 3650 rpm and 23 MPa shear force for 13 min to obtain a homogeneous aqueous matrix; immediately transfer the aqueous matrix to a vacuum degassing machine and degas it at -0.09 MPa pressure for 10 min. After degassing, the solid content of the aqueous matrix is ​​measured to be 6.5%.

[0051] S2: Cool the degassed aqueous matrix to 58°C at a rate of 2.5°C / min, add the formulated amount of compound moisturizing agent, ascorbate glucoside, and nicotinamide, start the stirrer and stir at 900 rpm for 11.5 min to dissolve, and observe every 2 min to ensure that no precipitation occurs, thus obtaining the primary mixture.

[0052] S3: Cool the primary mixture to 43°C at a rate of 1.5°C / min. First, add the prepared modified sheep placental small molecule protein and hydrolyzed fish collagen, and stir at 800 rpm for 5 min until completely dispersed. Then, add acetyl hexapeptide-8, glutathione, and hydrogenated modified ceramide NP in stepwise order, with an interval of 1.5 min between each two components. Finally, add sodium hyaluronate and panthenol, and continuously stir at 1350 rpm throughout the process. After all the core active components have been added, transfer the mixture to a low-frequency ultrasonic disperser, set the ultrasonic frequency to 25 kHz and the ultrasonic power to 125 W, and ultrasonically treat for 4 min. During the ultrasonic treatment, maintain the temperature of the mixture at 43°C using a constant temperature water bath. The particle size distribution of the core active mixture after ultrasonication was measured by a Malvern laser particle size analyzer and found to be 120-180 nm.

[0053] S4: Add the formulated amount of highly active soothing and repairing agent and alcohol-free compound preservative to the core active mixture. Adjust the pH of the system using a mixture of citric acid and arginine, first coarsely adjusting to 7.3, then finely adjusting to 6.0. Stir at 650 rpm during the adjustment process, and continue stirring for 6.5 minutes after adjustment. Filter the mixture through a 200-mesh nylon filter membrane at a filtration pressure of 0.3 MPa, collect the filtrate, and let it stand and mature at 25°C in the dark for 24 hours to obtain the finished skin care formula.

[0054] IV. Example 3 (a) Formula composition (by mass parts) 1. Core active ingredients: 2.5 parts of sheep placental small molecule protein, 0.15 parts of acetyl hexapeptide-8, and 0.08 parts of glutathione.

[0055] 2. Auxiliary active components: 4.0 parts of ascorbate glucoside, 1.2 parts of hydrogenated modified ceramide NP, 5.0 parts of nicotinamide, 3.0 parts of hydrolyzed fish collagen, 0.25 parts of sodium hyaluronate, and 2.0 parts of panthenol.

[0056] 3. Compound moisturizing agent: 12.0 parts.

[0057] 4. Highly active soothing and repairing agent: 0.2 parts.

[0058] 5. Alcohol-free compound preservative: 0.4 parts.

[0059] 6. Aqueous phase base materials: 0.15 parts xanthan gum, 0.12 parts carbomer.

[0060] (II) Preparation of small molecule proteins from sheep placenta 1. Degreasing and deodorizing pretreatment: Take fresh sheep placenta, rinse it 3 times with ultrapure deionized water, remove impurities such as fascia and blood vessels, and cut it into 1cm pieces. 3 Small pieces of placenta were placed in a high-speed homogenizer, and 3 times the mass of ultrapure deionized water were added. The homogenizer was homogenized at 10,000 rpm for 20 minutes to obtain a sheep placenta homogenate. 2 times the volume of ethanol was added to the homogenate, and the mixture was refluxed in a 50°C water bath for 1 hour. The precipitate was collected by centrifugation at 8,000 rpm for 20 minutes to remove fat. 0.5% activated carbon was added to the precipitate, and the mixture was stirred and adsorbed at 40°C for 30 minutes. The activated carbon was removed by filtration to obtain a degreased and deodorized sheep placenta tissue fluid.

[0061] 2. Stepwise enzymatic hydrolysis using a ternary complex protease: Papain, neutral protease, and bromelain were weighed at an enzyme activity ratio of 2:1:1, with the total enzyme addition amount being 1.5% of the sheep placental tissue fluid mass. First, the pH of the sheep placental tissue fluid was adjusted to 6.5, and the temperature was raised to 55℃. Papain was added, and the mixture was incubated for 2 hours, with stirring for 10 minutes every 30 minutes at a speed of 500 rpm. Then, the pH of the system was adjusted to 7.5, and the temperature was lowered to 50℃. A mixture of neutral protease and bromelain was added, and the enzymatic hydrolysis continued for 1 hour, with stirring as before. After the enzymatic hydrolysis was completed, the system was heated to 100℃ and incubated for 10 minutes to inactivate the enzyme. Then, it was cooled to room temperature, centrifuged at 8000 rpm for 20 minutes, and the supernatant was filtered through a 100-mesh filter to obtain sheep placental protein solution with a molecular weight of 1500-3500 Da.

[0062] 3. Phosphorylation modification: 5% sodium pyrophosphate was added to the sheep placental protein solution as a modifier. The mixture was stirred at 40°C for 3 hours at a speed of 300 rpm. After modification, the unreacted sodium pyrophosphate was removed by dialysis with a molecular weight cutoff of 1000 Da for 24 hours. After freeze-drying, the modified sheep placental small molecule protein was obtained. The degree of phosphorylation modification was found to be 50%.

[0063] (III) Skin Care Formula Preparation S1: Weigh the xanthan gum and carbomer according to the formula, add them to 69.98 parts of ultrapure deionized water, place them in an 84℃ constant temperature water bath, and homogenize them at 3800 rpm and 25 MPa shear force for 14 min to obtain a homogeneous aqueous matrix; immediately transfer the aqueous matrix to a vacuum degassing machine and degas it at -0.1 MPa pressure for 10 min. After degassing, the solid content of the aqueous matrix is ​​measured to be 8%.

[0064] S2: Cool the degassed aqueous matrix to 59°C at a rate of 3°C / min, add the formulated amount of compound moisturizing agent, ascorbate glucoside, and nicotinamide, start the stirrer and stir at 1000 rpm for 12 min to dissolve, observe every 2 min to ensure no precipitation occurs, and obtain the primary mixture.

[0065] S3: Cool the primary mixture to 44℃ at a rate of 2℃ / min. First, add the prepared modified sheep placental small molecule protein and hydrolyzed fish collagen, and stir at 800 rpm for 5 min until completely dispersed. Then, add acetyl hexapeptide-8, glutathione, and hydrogenated modified ceramide NP in stepwise order, with an interval of 1.5 min between each two components. Finally, add sodium hyaluronate and panthenol, and continuously stir at 1500 rpm throughout the process. After all the core active components have been added, transfer the mixture to a low-frequency ultrasonic disperser, set the ultrasonic frequency to 30kHz and the ultrasonic power to 150W, and ultrasonically treat for 5 min. During the ultrasonic treatment, maintain the temperature of the mixture at 44℃ using a constant temperature water bath. The particle size distribution of the core active mixture after ultrasonication was measured by a Malvern laser particle size analyzer and found to be 150-200nm.

[0066] S4: Add the formulated amount of highly active soothing and repairing agent and alcohol-free compound preservative to the core active mixture. Adjust the pH of the system using a mixture of citric acid and arginine, first coarsely adjusting to 7.5, then finely adjusting to 6.2. Stir at 700 rpm during the adjustment process, and continue stirring for 7 minutes after adjustment. Filter the mixture through a 200-mesh nylon filter membrane at a filtration pressure of 0.3 MPa, collect the filtrate, and let it stand and mature at 25°C in the dark for 24 hours to obtain the finished skin care formula.

[0067] V. Comparative Example 1 (a) Formula composition (by mass parts) 1. Core active components: 2.0 parts modified sheep placental small molecule protein, 0.12 parts acetyl hexapeptide-8, and 0.06 parts glutathione. The modified sheep placental small molecule protein undergoes defatting and deodorization pretreatment, followed by stepwise enzymatic hydrolysis with a binary complex protease. The binary complex protease consists of papain and neutral protease, with an enzyme activity ratio of 2:1 and a molecular weight of 2000-4000 Da.

[0068] 2. Auxiliary active components: 3.5 parts ascorbate glucoside, 1.0 part hydrogenated ceramide NP, 4.5 parts nicotinamide, 2.5 parts hydrolyzed fish collagen, 0.20 parts sodium hyaluronate, and 1.7 parts panthenol.

[0069] 3. Compound moisturizing agent: 10.0 parts, butylene glycol and 1,3-propanediol are compounded in a mass ratio of 2:1.

[0070] 4. Soothing and Repairing Agent: 0.15 parts, dipotassium glycyrrhizate and allantoin are compounded in a mass ratio of 2:1. The dipotassium glycyrrhizate has an activity content of 98% and the allantoin has a purity of 99%.

[0071] 5. Preservative: 0.3 parts, p-hydroxyacetophenone and 1,2-hexanediol are compounded in a mass ratio of 2:1. The antibacterial spectrum covers Gram-positive bacteria and Gram-negative bacteria, with a minimum inhibitory concentration of 0.1%-0.15%.

[0072] 6. Aqueous phase base materials: 0.12 parts xanthan gum, 0.10 parts carbomer.

[0073] (II) Preparation of modified sheep placental small molecule proteins 1. Degreasing and deodorizing pretreatment: Take fresh sheep placenta, rinse it 3 times with ultrapure deionized water, remove impurities such as fascia and blood vessels, and cut it into 1cm pieces. 3 Small pieces of placenta were placed in a high-speed homogenizer, and 3 times the mass of ultrapure deionized water were added. The homogenizer was homogenized at 10,000 rpm for 20 minutes to obtain a sheep placenta homogenate. 2 times the volume of ethanol was added to the homogenate, and the mixture was refluxed in a 50°C water bath for 1 hour. The precipitate was collected by centrifugation at 8,000 rpm for 20 minutes to remove fat. 0.5% activated carbon was added to the precipitate, and the mixture was stirred and adsorbed at 40°C for 30 minutes. The activated carbon was removed by filtration to obtain a degreased and deodorized sheep placenta tissue fluid.

[0074] 2. Stepwise enzymatic hydrolysis with binary complex protease: Papain and neutral protease were weighed at an enzyme activity ratio of 2:1, with the total enzyme addition amount being 1.5% of the mass of the sheep placental tissue fluid. First, the pH of the sheep placental tissue fluid was adjusted to 6.3, the temperature was raised to 53℃, papain was added, and the mixture was incubated for 2 hours, with stirring for 10 minutes every 30 minutes at a speed of 500 rpm. Then, the pH of the system was adjusted to 7.3, the temperature was lowered to 48℃, neutral protease was added, and the enzymatic hydrolysis continued for 1 hour, with the stirring method as before. After the enzymatic hydrolysis was completed, the system was heated to 100℃ and incubated for 10 minutes to inactivate the enzyme. Then, it was cooled to room temperature, centrifuged at 8000 rpm for 20 minutes, and the supernatant was filtered through a 100-mesh filter to obtain a modified sheep placental small molecule protein solution with a molecular weight of 2000-4000 Da, which was then freeze-dried for later use.

[0075] (III) Skin Care Formula Preparation S1: Weigh the xanthan gum and carbomer according to the formula, add them to 73.75 parts of ultrapure deionized water, place them in an 83℃ constant temperature water bath, and homogenize them at 3650 rpm and 23 MPa shear force for 13 min to obtain a homogeneous aqueous matrix; immediately transfer the aqueous matrix to a vacuum degassing machine and degas it at -0.07 MPa pressure for 10 min. After degassing, the solid content of the aqueous matrix is ​​measured to be 6.5%.

[0076] S2: Cool the degassed aqueous matrix to 58°C at a rate of 2.5°C / min, add the formulated amount of compound moisturizing agent, ascorbate glucoside, and nicotinamide, start the stirrer and stir at 900 rpm for 11.5 min to dissolve, and observe every 2 min to ensure that no precipitation occurs, thus obtaining the primary mixture.

[0077] S3: Cool the primary mixture to 43°C at a rate of 1.5°C / min. First, add the prepared modified sheep placental small molecule protein and hydrolyzed fish collagen, and stir at 800 rpm for 5 min until completely dispersed. Then, add acetyl hexapeptide-8, glutathione, and hydrogenated modified ceramide NP in stepwise order, with an interval of 1.5 min between each two components. Finally, add sodium hyaluronate and panthenol, and continuously stir at 1350 rpm throughout the process. After all the core active components have been added, transfer the mixture to a low-frequency ultrasonic disperser, set the ultrasonic frequency to 15 kHz and the ultrasonic power to 80 W, and ultrasonically treat for 4 min. During the ultrasonic treatment, maintain the temperature of the mixture at 43°C using a constant temperature water bath. The particle size distribution of the core active mixture after ultrasonication was measured by a Malvern laser particle size analyzer and found to be 200-300 nm.

[0078] S4: Add the formulated amount of soothing and repairing agent and preservative to the core active mixture. Adjust the pH of the system using a mixture of citric acid and arginine, first coarsely adjusting to 7.3, then finely adjusting to 6.0. Stir at 650 rpm during the adjustment process, and continue stirring for 6.5 minutes after adjustment. Filter the mixture through a 180-mesh nylon filter membrane at a filtration pressure of 0.3 MPa. Collect the filtrate and let it stand and mature at 25°C in the dark for 24 hours to obtain the finished skin care formula.

[0079] VI. Performance Testing and Result Analysis (a) Basic performance testing 1. Stability Test: The finished products of Examples 1-3 and Comparative Example 1 were placed in environments of 4℃, 25℃, and 45℃ respectively, and left to stand for 3 months. The effects of stratification, precipitation, and odor were observed. The results showed that Examples 1-3 did not exhibit stratification, precipitation, or obvious odor under any temperature conditions; Comparative Example 1 showed slight stratification after being placed at 45℃ for 2 months, but no obvious odor.

[0080] 2. Particle size and Zeta potential test: The particle size D90 of the finished products of Examples 1-3 is ≤200nm and the absolute value of Zeta potential is ≥35mV, indicating good system stability; the particle size D90 of Comparative Example 1 is 280nm and the absolute value of Zeta potential is 30mV, indicating good system stability.

[0081] 3. Mildness Test: Thirty volunteers with sensitive skin were selected and a patch test was performed on the inside of their arms for 7 consecutive days. Volunteers in Examples 1-3 showed no redness, swelling, itching, or other discomfort, and their irritation scores were all 0. In Comparative Example 1, three volunteers experienced mild redness without itching, and their irritation scores were 0.

[0082] (ii) Efficacy testing 1. Anti-aging performance: After using the product for 28 days, the skin collagen content was tested. The skin collagen content of Examples 1-3 increased by 28%, 35%, and 32%, respectively; the skin collagen content of Comparative Example 1 increased by 22%.

[0083] 2. Fine line reduction performance: The depth of fine lines on the skin was measured using a skin roughness meter after 42 days of use. The fine line depth of Examples 1-3 was reduced by 25%, 30%, and 28%, respectively; the fine line depth of Comparative Example 1 was reduced by 18%.

[0084] 3. Brightening performance: The skin tone L value was measured using a skin tone meter after 28 days of use. The L values ​​of Examples 1-3 increased by 3.2, 4.0 and 3.8 respectively; the L value of Comparative Example 1 increased by 2.5.

[0085] (III) Anti-corrosion and moisture retention performance test 1. Anti-corrosion performance: The antibacterial rate of the finished product against Gram-positive bacteria, Gram-negative bacteria and mold was tested. The antibacterial rate of Examples 1-3 against the three types of microorganisms was ≥99%; the antibacterial rate of Comparative Example 1 against mold was 92%, and the antibacterial rate against Gram-positive bacteria and Gram-negative bacteria was ≥99%.

[0086] 2. Moisturizing performance: The skin moisture content was measured 4 hours after use using a skin moisture meter. The skin moisture content of Examples 1-3 remained at 85%, 88%, and 86% of the initial value, respectively; the skin moisture content of Comparative Example 1 remained at 78% of the initial value.

[0087] In summary, Examples 1-3, through stepwise enzymatic hydrolysis and phosphorylation modification of ternary complex protease to optimize the performance of sheep placental protein, combined with a scientifically formulated moisturizing, soothing, and preservative system and precise preparation process, are superior to Comparative Example 1 in terms of stability, gentleness, preservative and moisturizing effects, and core anti-aging, wrinkle-reducing, and brightening effects, fully demonstrating the technical advantages of the present invention. Although Comparative Example 1 is slightly inferior in some aspects, its overall performance is good and no obvious shortcomings are found.

[0088] The basic performance comparison between the examples and the comparative examples is shown in the table below: Table 1 The core efficacy and specific performance of the examples and comparative examples are compared in the table below: Table 2 The two tables systematically compared the basic performance and core efficacy of each embodiment and the comparative example. The results showed that the embodiments performed better in terms of stability, utilization of active ingredients, anti-aging, wrinkle reduction and brightening, and had better moisturizing and anti-corrosion properties. The comparative example had good overall performance without obvious shortcomings, but its overall performance was not as good as that of the embodiments, which confirmed the advantages of the technical solution of the present invention.

[0089] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. An animal placental protein anti-wrinkle and skin lightening formulation, characterized in that, By mass parts of core active ingredient and auxiliary active ingredient compounded, the core active ingredient is: 1.5-2.5 parts of sheep placenta small molecular protein, 0.08-0.15 parts of acetyl hexapeptide-8, 0.04-0.08 parts of glutathione, the sheep placenta small molecular protein is pretreated by degreasing and deodorization, step-by-step enzymolysis by three-element complex protease, and phosphorylation modification, and the modified sheep placenta small molecular protein has a molecular weight of 1500-3500 Da and a phosphorylation modification degree of 30%-50%; the auxiliary active ingredient is: 3.0-4.0 parts of ascorbic acid glucoside, 0.8-1.2 parts of hydrogenated modified ceramide NP, 4.0-5.0 parts of nicotinamide, 2.0-3.0 parts of hydrolyzed fish collagen, 0.15-0.25 parts of sodium hyaluronate, and 1.5-2.0 parts of panthenol.

2. The animal placental protein anti-wrinkle and skin lightening formulation according to claim 1, wherein The compound moisturizing aid further includes 8.0-12.0 parts of a compound moisturizing aid, which is a compound of butanediol, 1,3-propanediol, and polyoxyethylene ether modified glycerol in a mass ratio of 3:2:

1.

3. The animal placental protein anti-wrinkle and skin lightening formulation according to claim 1, wherein The three-element complex protease is composed of papain, neutral protease, and bromelain, and the enzyme activity ratio is 2:1:

1.

4. The animal placental protein anti-wrinkle and skin lightening formulation according to claim 1, wherein The high-activity soothing agent further includes 0.1-0.2 parts of a high-activity soothing agent, which is a compound of dipotassium glycyrrhizinate, allantoin, and a panthenol derivative, pyrrolidone carboxylic acid zinc, in a mass ratio of 2:1:1, wherein the active content of dipotassium glycyrrhizinate is ≥98%, and the purity of allantoin is ≥99%.

5. The animal placental protein anti-wrinkle and skin lightening formulation according to claim 1, wherein The non-alcohol compound preservative further includes 0.2-0.4 parts of a non-alcohol compound preservative, which is a compound of p-hydroxyacetophenone, caprylhydroxamic acid, 1,2-hexanediol, and ethylhexylglycerin in a mass ratio of 3:2:2:1, and the preservative system covers gram-positive bacteria, gram-negative bacteria, and mold in the antibacterial spectrum, and the minimum inhibitory concentration is 0.05%-0.1%.

6. The animal placental protein anti-wrinkle and skin lightening formulation according to claim 3, wherein The process parameters of the step-by-step enzymolysis of the three-element complex protease are as follows: first, enzymolysis by papain at 50-55°C and pH 6.0-6.5 for 2h, then continuous enzymolysis by a mixed enzyme of neutral protease and bromelain at 45-50°C and pH 7.0-7.5 for 1h, enzyme inactivation at 100°C for 10min, centrifugal filtration to obtain sheep placenta protein liquid with a molecular weight of 1500-3500 Da, and phosphorylation modification using sodium pyrophosphate as a modifier at a modification temperature of 40°C for 3h.

7. A method of preparing an anti-wrinkle and skin lightening formulation of animal placental protein, characterized in that, The method comprises the following steps: S1, a formula amount of water phase basic auxiliary material is added to ultra-pure deionized water with an electric conductivity of ≤10 μS / cm, and a uniform water phase matrix is prepared by heating in a 82-84°C water bath and homogenizing and shearing at a speed of 3500-3800 rpm for 12-14 min; S2, the water phase matrix is cooled to 57-59°C at a rate of 2-3°C / min, a formula amount of compound moisturizing aid, ascorbic acid glucoside, and nicotinamide is added, and a primary mixed solution without precipitation is prepared by stirring and dissolving at a speed of 800-1000 rpm for 11-12 min; S3, the primary mixed solution is cooled to 42-44℃ at a rate of 1-2℃ / min, and the modified sheep placenta small molecular protein and the hydrolyzed fish collagen are first added and stirred for 5 min, then the acetyl hexapeptide-8, glutathione and hydrogenated modified ceramide NP are added in sequence, and finally the sodium hyaluronate and panthenol are added, and the whole process is continuously stirred at a speed of 1200-1500 rpm to prepare a core active mixed solution; S4, the high-activity soothing repair agent and the alcohol-free compound preservative and preservative are added to the core active mixed solution, the pH of the system is adjusted to 5.8-6.2 using a mixed solution of citric acid and arginine, and the stirring speed is 600-700 rpm during the adjustment process. After adjustment, continue to stir for 6-7 min, filter through a filter membrane to prepare a skin care formula finished product.

8. The method of claim 7, wherein the animal placental protein anti-wrinkle and skin lightening formulation is prepared by the steps of: The water phase matrix prepared in S1 needs to be immediately subjected to vacuum degassing treatment, the degassing pressure is-0.08~-0.1MPa, the degassing time is 10min, the solid content of the water phase matrix after degassing is 5%-8%, and the shear force of the homogeneous shear is controlled to be 20-25MPa.

9. The method of claim 7, wherein the animal placental protein anti-wrinkle and skin lightening formulation is prepared by the steps of: The interval of the step-by-step addition of each peptide, protein and ceramide component in S3 is 1.5 min, the citric acid and arginine mixture in S4 is compounded at a mass ratio of 1:2, the pH adjustment uses a gradient adjustment method, first coarsely adjusted to 7.0-7.5 and then finely adjusted to 5.8-6.2, the filter membrane is a 200 mesh nylon filter membrane, and the finished product is placed at 25℃ in the dark for 24h.

10. The method of claim 7, wherein the animal placental protein anti-wrinkle and skin lightening formulation is prepared by the steps of: After the addition of all the core active components in S3 is completed, the core active mixed solution is subjected to low-frequency ultrasonic dispersion treatment, the ultrasonic frequency is 20-30kHz, the ultrasonic power is 100-150W, and the ultrasonic time is 3-5min, the particle size distribution of the core active mixed solution after ultrasonic treatment is 100-200nm, and the temperature of the mixed solution is maintained at 42-44℃ during the ultrasonic process.

Images