A recombinant albumin anti-aging composition, its preparation method and application

Abstract

The present invention relates to the field of cosmetics, and specifically relates to a recombinant albumin anti-aging composition, a preparation method thereof, and an application. The composition of the present invention comprises 0.1 to 15.0 parts by mass of recombinant human albumin; 0.1 to 10.0 parts of wine lees polysaccharide extract; and 0.1 to 5.0 parts of raspberry extract. In the composition disclosed by the present invention, the recombinant human albumin, the wine lees polysaccharide extract, and the raspberry extract act synergistically, have the effect of inhibiting the activity of tyrosinase, promoting the regeneration of collagen and elastin, have the effect of firming and anti-wrinkle, can effectively resist aging, and the composition of the present invention has simple components, good biocompatibility, improves the sensitization of the wine lees extract and the raspberry extract, and can be widely applied to the preparation of anti-aging products.

Description

Technical Field

[0001] The present invention relates to the field of cosmetics, and particularly to a recombinant albumin anti-aging composition, a preparation method thereof, and an application thereof. Background Art

[0002] With the increase of age, factors such as lifestyle and sun exposure lead to a decrease in skin collagen, resulting in facial skin aging, dullness, loss of elasticity, the appearance of spots, wrinkles, etc. Related cosmetics emerge in an endless stream, and masks have become an essential thing in people's daily lives due to their convenience and effectiveness. However, the active ingredients of most masks on the market are prepared by chemical synthesis, which may produce certain effects in the short term, but long-term use will endanger human health.

[0003] Wine lees: Wine lees refer to the grape residues filtered during the wine brewing process. The lees are mainly composed of dead yeast cells, grape seeds, grape skins, grape stalks, and insoluble tartaric acid, which are usually precipitated during the wine brewing and aging processes. The lees are rich in protein amino acids, active polypeptide classes, functional oligosaccharides, antioxidant phenols and flavonoids, and also contain other functional components and various trace elements.

[0004] Red raspberry extract: Red raspberry (Rubus idaeus L.) is a perennial small shrub fruit tree of the genus Rubus in the family Rosaceae, also known as wood raspberry, mountain raspberry, and raspberry. Red raspberry originated in Europe, and Russia is the country where red raspberry was cultivated earliest. Currently, it is widely planted in Europe, Asia, and the Americas. It contains rich nutrients and bioactive components, such as sugars, vitamins, minerals, anthocyanins, ellagic acid, raspberry ketone, etc., and has various effects such as antioxidant, antibacterial, anti-aging, anti-inflammatory, anti-cancer, improving immunity, protecting the heart, and losing weight, and has a wide range of applications in the fields of food, medicine, health products, etc.

[0005] However, due to their sensitization, wine lees extract and red raspberry extract are more applied in the fields of health products or food, and there is currently relatively little research in the field of cosmetics. Summary of the Invention

[0006] In view of the deficiencies in the prior art, the present invention applies wine lees and red raspberry to the field of cosmetics, and provides an anti-aging composition, a preparation method thereof, and an application thereof. The specific content of the present invention is as follows:

[0007] In the first aspect of the present invention, an anti-aging composition is provided, and the composition includes: 0.1 to 15.0 parts by mass of recombinant human albumin; 0.1 to 10.0 parts of wine lees polysaccharide extract; 0.1 to 5.0 parts of red raspberry extract.

[0008] Furthermore, the composition further comprises additives acceptable in the field of cosmetics. Further, the additives include at least one of preservatives, emulsifiers, humectants, thickeners, pH regulators, surfactants, stabilizers, antioxidants, and emollients.

[0009] Further, the additives include at least one of phenoxyethanol, parabens, vitamin E (tocopherol), butylated hydroxyanisole (BHA), dibutylhydroxytoluene (BHT), polysorbates (Tween series), glyceryl stearate, carbomer, xanthan gum, glycerol, citric acid, sodium hydroxide, sodium lauryl sulfate (SLS), cocamidopropyl betaine, ethylenediaminetetraacetic acid (EDTA), and ethylhexyl palmitate.

[0010] In a specific embodiment of the present invention, the additives include glycerol, carbomer, ethylhexyl palmitate, and polysorbate 80.

[0011] It should be noted that the present invention does not strictly limit the components and mass fractions of the additives acceptable in the field of cosmetics. All additives having the functions of preservatives, emulsifiers, humectants, thickeners, pH regulators, surfactants, stabilizers, antioxidants, and emollients that are harmless to the human body and selected according to the dosage form of the product, or the routine adjustment of the mass fractions of the additives according to actual needs, are within the scope of protection of the present invention.

[0012] Further, the dosage form of the composition includes at least one of aqueous solutions, emulsions, essences, creams, ointments, gels, facial masks, and powders. In a specific embodiment of the present invention, the dosage form of the composition is a freeze-dried facial mask.

[0013] Further, the composition can effectively inhibit the activity of tyrosinase, promote the regeneration of collagen and elastin, thereby having the effect of firming and anti-wrinkle, and can significantly resist aging.

[0014] In the second aspect of the present invention, there is provided the application of the described composition in any of the following aspects:

[0015] A1: Application in the preparation of anti-aging products;

[0016] A2: Application in the preparation of firming and anti-wrinkle products;

[0017] A3: Application in the preparation of products for promoting the regeneration of collagen and / or elastin;

[0018] A4: Application in the preparation of products for inhibiting the activity of tyrosinase.

[0019] Further, the anti-aging product, firming and anti-wrinkle product, product for promoting collagen and / or elastin regeneration, or product for inhibiting tyrosinase activity includes cosmetics.

[0020] Further, the composition of the present invention is an anti-aging freeze-dried mask, and the freeze-dried mask includes: recombinant human albumin in an amount of 0.1 to 15.0 parts by mass; wine lees polysaccharide extract in an amount of 0.1 to 10.0 parts by mass; raspberry extract in an amount of 0.1 to 5.0 parts by mass; humectant in an amount of 5.0 to 10.0 parts by mass; thickener in an amount of 0.1 to 0.8 parts by mass; emollient in a mass fraction of 0.3 to 1.0 parts; stabilizer in a mass fraction of 0.01 to 0.20 parts.

[0021] In a specific embodiment of the present invention, the anti-aging freeze-dried mask includes: recombinant human albumin in an amount of 0.1 to 15.0 parts by mass; wine lees polysaccharide extract in an amount of 0.1 to 10.0 parts by mass; raspberry extract in an amount of 0.1 to 5.0 parts by mass; glycerol in an amount of 5.0 to 10.0 parts by mass; carbomer in an amount of 0.1 to 0.8 parts by mass; ethylhexyl palmitate in a mass fraction of 0.3 to 1.0 parts; polysorbate 80 in a mass fraction of 0.01 to 0.20 parts.

[0022] In the third aspect of the present invention, a method for preparing the composition is provided, and the method includes the following steps:

[0023] S1: Preparation of wine lees polysaccharide extract: After sterilizing, drying, pulverizing, and sieving through a 10-mesh sieve the wine lees, a crude extract powder of wine lees polysaccharide is obtained. The wine lees polysaccharide is extracted by the method of water extraction and alcohol precipitation. It is soaked with deionized water at a mass ratio of the crude extract to deionized water of 1:(3 - 7) for extraction, and the extract is concentrated and then allowed to stand and precipitate with an ethanol solution with a concentration of more than 85% (v / v). After centrifugation, it is suspended with purified water and then spray-dried to obtain the wine lees polysaccharide extract. Preferably, the extraction step can be repeated 2 to 5 times, and the filtrates from multiple extractions are combined and then concentrated;

[0024] S2: Preparation of raspberry extract: The dried raspberries are pulverized and sieved through a 10-mesh sieve to obtain a powder, which is soaked in 75% - 80% (v / v) ethanol for 6 - 8 h, the ethanol is distilled off, and then it is suspended with deionized water and spray-dried to obtain a spray-dried powder of raspberry extract;

[0025] S3: The spray-dried powder of wine lees polysaccharide extract and the spray-dried powder of raspberry extract in parts by mass are added to an aqueous solution or salt buffer of recombinant human albumin in parts by mass, and high-pressure homogenization is carried out using a high-pressure homogenizer at 500 - 1000 bar for 5 - 15 min at a temperature of 35°C - 60°C to obtain a mixed solution of the anti-aging composition.

[0026] Further, the conditions for spray drying in steps S1 and S2 are as follows: inlet air temperature: 120~160°C; atomization pressure: 0.1~1.0 Mpa; feeding rate: 200~800 ml / h.

[0027] Further, the method for sterilizing wine distillers' grains in step S1 is high-pressure sterilization at a temperature of 115~125°C for 15~30 min.

[0028] Further, the specific steps for centrifuging the polysaccharide extract from wine distillers' grains in step S1 are: 5000~8000 r / min for 15~30 min.

[0029] Further, the method further includes a step of adding additives acceptable in the cosmetic field to the anti-aging composition mixed solution obtained in step S3 and mixing them evenly.

[0030] In a specific embodiment of the present invention, the method for preparing the recombinant albumin anti-aging composition includes the following steps:

[0031] (1) The polysaccharide extract from wine distillers' grains is prepared as follows: subject the wine distillers' grains to high-pressure sterilization (121°C, 15~30 min), take the sterilized wine distillers' grains for drying, crush the dried wine distillers' grains after drying, pass through a 10-mesh sieve to obtain a powder, extract with deionized water, soak at a solid-liquid ratio of 1:5 for 1 h, reflux and extract for 15 min at a water bath temperature, extract 3 times, combine the filtrates and concentrate, add 3 times the amount of 95% ethanol to the concentrated solution, place in the refrigerator and let stand for 24 h, centrifuge the polysaccharide precipitate obtained by alcohol precipitation at 5000~8000 r / min for 15~30 min, take the precipitate, suspend it in purified water, and spray dry the suspended solution to obtain it;

[0032] (2) The raspberry extract is prepared as follows: crush the dried raspberries, pass through a 10-mesh sieve to obtain a powder, soak in 75%~80% (v / v) ethanol for 6~8 h, distill off the ethanol, then suspend it in deionized water, and spray dry to obtain the spray-dried powder of the raspberry extract;

[0033] (3) Inlet air temperature for spray drying: 120~160°C, preferably 150°C; atomization pressure: 0.1~1.0 Mpa, preferably 0.5 Mpa; feeding rate: 200~800 ml / h, preferably 450 ml / h;

[0034] (4) Take an appropriate amount of deionized water, add the spray-dried powder of the polysaccharide extract from wine distillers' grains and the spray-dried powder of the raspberry extract in parts by mass to the recombinant human albumin in parts by mass, and use a high-pressure homogenizer at 500~1000 bar for high-pressure homogenization for 5~15 min at a temperature of 35°C~60°C to obtain a composition solution.

[0035] The beneficial effects of the present invention include but are not limited to:

[0036] 1. In the composition of the present invention, recombinant human albumin, polysaccharide extract from wine lees, and raspberry extract act synergistically to inhibit the activity of tyrosinase, promote the regeneration of collagen and elastin, have the effect of firming and anti-wrinkle, can effectively resist aging, and the composition of the present invention has simple components, good biocompatibility, no obvious irritation and sensitization to the human body, and can be widely used in the preparation of anti-aging products.

[0037] 2. In the composition of the present invention, recombinant human albumin effectively improves the sensitization of polysaccharide extract from wine lees and raspberry extract, and expands the application of polysaccharide extract from wine lees and raspberry extract in the field of cosmetics.

[0038] 3. The preparation method of the composition of the present invention has a simple production process and has high application prospects. Detailed Embodiments

[0039] The present invention will be described in detail below with reference to the embodiments. However, the present invention is not limited to these embodiments. Unless otherwise specified, the raw materials and catalysts in the embodiments of the present invention are purchased through commercial channels.

[0040] Definition: Recombinant human albumin refers to albumin prepared by using recombinant DNA technology for treatment and prevention.

[0041] Source of materials: Recombinant human albumin: Tonghua Anruite Biopharmaceutical Co., Ltd., product number: ART101S;

[0042] Wine lees: Provided by Tonghua Wine Co., Ltd.,

[0043] Raspberry: Provided by Tonghua Huinan Raspberry Industry Base,

[0044] Equipment:

[0045] Water bath (HH.S21-8, Shanghai Boxun Medical Biological Instrument Co., Ltd.);

[0046] Multifunctional fluorescence microplate reader (SPMax 3500FL type, Shanghai Flash Spectrum Biotechnology Co., Ltd.);

[0047] Analytical balance (BSA224S type, Sartorius Scientific Instruments Beijing Co., Ltd.);

[0048] Tyrosinase: (Batch number: T885829-100KU, Shanghai Macklin Biochemical Co., Ltd.);

[0049] Professional skin tester: (Model: CBS-805, CBS Optoelectronics Co., Ltd., Taiwan)

[0050] Hydrogel mask base fabric (Batch number: 20231102, Shanghai Fulei New Materials Technology Co., Ltd.);

[0051] pH meter (Model PB-10, Sartorius Scientific Instruments Beijing Co., Ltd.);

[0052] Other reagents and instruments used are all commercially available, unless otherwise specified.

[0053] Example 1: Preparation of a recombinant human albumin anti-aging composition containing the following components in parts by mass: The active ingredients are shown in Table 1.

[0054] Table 1 Composition table of various recombinant human albumin anti-aging freeze-dried masks

[0055]

[0056] Table 2 Control group information table

[0057]

[0058] The specific steps for preparing the recombinant human albumin anti-aging composition are as follows:

[0059] (1) The polysaccharide extract of wine lees is prepared by the following method: Autoclave the wine lees at 121 °C for 15 - 30 min, take the sterilized wine lees for drying, crush the dried wine lees after drying, pass through a 10-mesh sieve to obtain a powder, extract with deionized water, soak for 1 h according to a solid-liquid ratio of 1:5, reflux and extract for 15 min at a water bath temperature, extract 3 times, combine the filtrates and concentrate. Add 3 times the amount of 95% ethanol to the concentrated solution and place it in the refrigerator for 24 h. Centrifuge the polysaccharide precipitate obtained by alcohol precipitation at 5000 - 8000 r / min for 15 - 30 min. Take the precipitate and suspend it in purified water, and spray-dry the suspended solution to obtain it.

[0060] (2) The raspberry extract is prepared by the following method: Crush the dried raspberries, pass through a 10-mesh sieve to obtain a powder, soak in 75% - 80% (v / v) ethanol for 6 - 8 h, distill off the ethanol, then suspend it in deionized water, and spray-dry to obtain the spray-dried powder of raspberry extract;

[0061] (3) Inlet air temperature for spray drying: 120 - 160 °C, preferably 150 °C; Atomization pressure: 0.1 - 1.0 Mpa, preferably 0.5 Mpa; Feed rate: 200 - 800 ml / h, preferably 450 ml / h.

[0062] (4) Take an appropriate amount of deionized water, spray-dried polysaccharide extract from wine distillers' grains and spray-dried raspberry extract in parts by mass, add them to recombinant human albumin in parts by mass, and use a high-pressure homogenizer at 500 - 1000 bar for high-pressure homogenization for 5 - 15 min at a temperature of 35°C - 60°C to obtain a composition solution.

[0063] Test Example 1: Tyrosinase Activity Inhibition Test

[0064] Take 1 ml of the composition solution, 0.9 ml of tris(hydroxymethyl)aminomethane (Tris) hydrochloride buffer with a pH of 7.6, and 1 ml of tyrosine with a mass fraction of 0.03% in a test tube, place it in a constant temperature water bath at 37°C for 10 min, add 0.1 ml of tyrosinase aqueous solution (1070 U / ml), after mixing the solution evenly, place it in a constant temperature water bath at 37°C for 30 min, and measure its absorbance A at 475 nm using a UV spectrophotometer. 供试品

[0065] Take 1 ml of Tris buffer, 0.9 ml of tris(hydroxymethyl)aminomethane (Tris) hydrochloride buffer with a pH of 7.6, and 1 ml of tyrosine with a mass fraction of 0.03% in a test tube, place it in a constant temperature water bath at 37°C for 10 min, add 0.1 ml of tyrosinase aqueous solution (1070 U / ml), after mixing the solution evenly, place it in a constant temperature water bath at 37°C for 30 min, and measure its absorbance A at 475 nm using a UV spectrophotometer. 阴性

[0066] Take 1 ml of the composition solution, 0.9 ml of tris(hydroxymethyl)aminomethane (Tris) hydrochloride buffer with a pH of 7.6, and 1 ml of tyrosine with a mass fraction of 0.03% in a test tube, place it in a constant temperature water bath at 37°C for 10 min, add 0.1 ml of tris(hydroxymethyl)aminomethane (Tris) hydrochloride buffer with a pH of 7.6, after mixing the solution evenly, place it in a constant temperature water bath at 37°C for 30 min, and measure its absorbance A at 475 nm using a UV spectrophotometer. 空白 Calculate the inhibition rate of the test sample on tyrosinase. The data results are shown in Table 3.

[0067] Inhibition rate (%) = [ (A 阴性 - A 空白 ) - (A 供试品 - A 空白 ) / (A 阴性 - A 空白 ) × 100%

[0068] Table 3 Experimental Results of Tyrosinase Activity Inhibition Test

[0069]

[0070] As can be seen from the data in Table 3, the experimental results of the recombinant human albumin anti-aging composition on tyrosinase inhibition showed that the tyrosinase inhibition rate was above 69%, indicating that the prepared recombinant human albumin anti-aging composition had an inhibitory effect on tyrosinase.

[0071] For the experimental groups (groups 14 - 16) that only added single components of recombinant human albumin, wine lees polysaccharide extract, and raspberry extract, the experimental data was compared with the experimental groups (groups 1 - 13, groups 17 - 18). The tyrosinase inhibition rate was the lowest, indicating that the single component had the weakest inhibitory ability on tyrosinase. At the same time, the data results of the experimental groups with the combination of two functional components (groups 11 - 13) were much lower than those of groups 1 - 7, indicating that only when the three active components coexisted could this anti-aging facial mask liquid maintain a strong enzyme inhibition rate. For the optimal formulation ratio, the experimental results of the experimental groups (groups 1 - 7) were higher than those of the soy protein group, bovine serum albumin group, and ovalbumin group, indicating that the inhibitory effect of recombinant human albumin on tyrosinase was better than that of serum albumin from plants and animals. Compared with other compatibility groups (groups 8 - 18), the tyrosinase inhibition rate > 93%. At the same time, compared with the blank control group (group 19), group 4 had a significant difference, indicating that the recombinant human albumin anti-aging composition prepared by this method had a significant inhibitory effect on tyrosinase and could play an anti-aging role.

[0072] Test Example 2: Human Skin Irritation Test

[0073] According to the "Technical Specifications for Cosmetics Safety" (2015 version), a human patch test was conducted. 45 males and 45 females aged 20 - 30 (without skin diseases and iatrogenic skin sensitivity) were selected as subjects and randomly divided into nine groups, with 5 males and 5 females in each group for the patch test. The patch test was carried out on the inner side of the forearm and behind the ear of the subjects. After 30 minutes of the test, the test patch was removed. The compositions prepared by the experimental groups (groups 1, 4, 7, 11, 12, 13, 14, 15, 16) were selected for testing. After removing the test patch at 0.5, 12, and 24 hours after the test, the skin reactions were observed after the test was completed, and the evaluation was made with reference to Table 4.

[0074] Table 4 Skin Patch Test Evaluation Criteria

[0075]

[0076] The experimental results showed that in the patch test on the skin of the subjects, among the subjects in Group 12, 4 showed weak positive reactions at 24 hours, and the symptoms disappeared after stopping the use; among the subjects in Groups 14 and 15, 2 showed weak positive reactions at 24 hours, and the symptoms disappeared after stopping the use; among the subjects in Groups 11 and 13, 1 showed a suspicious erythema reaction; the patch test results of the subjects in Groups 1, 4, and 7 were all negative. Through the above experimental results, it was shown that using Groups 12, 14, and 15 might cause sensitization, especially the probability of sensitization by wine lees polysaccharide extract and raspberry extract was higher, indicating that the body's immune system of the subjects had a certain degree of immune response to the test substances, but the reaction degree was relatively weak, and the corresponding symptoms disappeared immediately after stopping the use; in Group 11, 1 showed a suspicious erythema reaction; the subjects using the combination of recombinant human albumin, raspberry extract and wine lees polysaccharide extract all showed negative reactions. According to the evaluation system of the patch test, negative results indicated that under the current experimental conditions, the skin of the subjects in these groups did not show detectable immune reactions or inflammatory reactions to the test substances in contact. It was verified by the negative patch test results of Groups 1, 4, and 7, indicating that the designed composition might have changed the originally allergy-causing characteristics of wine lees polysaccharide extract and raspberry extract, and the encapsulation effect produced by recombinant human albumin or specific binding to certain receptors of recombinant human albumin might reduce the sensitization of wine lees extract and raspberry extract.

[0077] Based on the composition prepared by the invention patent, in the actual application scenario simulated in this experiment, it has no obvious irritation and sensitization to human skin. From the perspective of safety evaluation indicators, this composition has good safety performance.

[0078] Test Example 3: Anti-wrinkle and firming experiment

[0079] Elastin and collagen have the functions of preventing skin aging and promoting regeneration, making the skin elastic and firm. A CBS-805 professional skin tester was used to detect elastin and collagen.

[0080] According to the "Technical Specifications for Cosmetics Safety" (2015 edition), 90 Chinese women aged 35 - 65 were selected for subject screening and randomly divided into three groups, and the compositions prepared by the experimental groups (groups 1, 4, and 7) were used respectively. The subjects used it once daily after cleansing their faces, and evenly applied the corresponding composition on the cheek area of the face. Other anti-wrinkle products were discontinued during the experiment. Skin tests were conducted at the 0th week, 2nd week, 4th week, and 8th week. Before the test, the facial skin was cleaned with water, and after cleaning, the subjects sat quietly in a constant temperature and humidity room for 30 minutes. After 30 minutes, elastin and collagen tests were conducted on the subjects participating in the experiment, and the control was the self-before-and-after comparison. The elastin and collagen test data of all subjects before using the product were used as the control group indicators, and the indicators of the subjects after using the product for 2 weeks, 4 weeks, and 8 weeks were used as the experimental group indicators. The experimental results are shown in Tables 5 and 6.

[0081] Table 5 Elastin Test Results

[0082]

[0083] Note: Compared with the 0th week, P≤0.05, * indicates significant difference.

[0084] According to the indicators of the CBS-805 professional skin tester, the elastin value of the subjects was detected. The higher the value, the firmer the skin. The detection value of this instrument is in the range of 59 - 70, and the skin state is firm. According to the test results in Table 5: The content of elastin in the subjects using groups 1, 4, and 7 increased significantly after 2 weeks, 4 weeks, and 8 weeks of testing. Among them, the effect of group 4 was significantly higher than that of the subjects using group 1 and group 7, indicating that at different time points, group 4 had a more significant effect on increasing the elastin value. However, although the elastin of the subjects in groups 11 - 16 using single components or two-component combinations also increased, the effect was significantly lower than that of groups 1, 4, and 7 with three-component combination and synergy, indicating that the recombinant human albumin, wine lees extract, and raspberry extract in the anti-aging composition prepared by this method have a specific synergistic effect and can significantly promote elastin regeneration.

[0085] Table 6 Collagen Test Results

[0086]

[0087] Note: Compared with the 0th week, P≤0.05, * indicates significant difference.

[0088] According to the indicators of the CBS-805 professional skin tester, when the detected value is in the range of 25-50, there is a lack of collagen fibers; when it is 50-65, the collagen fibers are sparse; and when it is greater than 65, the collagen fibers are normal. According to the test results in Table 6: in terms of the collagen index, the content of collagen protein in the subjects of usage groups 1, 4, and 7 increased significantly after 2 weeks, 4 weeks, and 8 weeks of testing. Among them, the collagen content of the subjects in group 4 was significantly higher than that of the subjects in usage groups 1 and 7, indicating that the combination method adopted in group 4 has a more prominent effect in enhancing the collagen content. However, although the elastin in the subjects of usage groups 11-16 using single components or pairwise combinations also increased, the effect was significantly lower than that of groups 1, 4, and 7 with three-component synergy, indicating that the recombinant human albumin, grape wine lees extract, and raspberry extract in the anti-aging composition prepared by this method have a specific synergistic effect, which can effectively increase the collagen content in the skin, thereby improving the collagen fiber state of the skin and helping to maintain the normal structure and function of the skin.

[0089] In summary, the anti-aging composition prepared by this method can significantly promote the regeneration of collagen and elastin, increase the content of collagen and elastin, and has the effects of anti-wrinkle and firming.

[0090] Example 2: Preparation of a freeze-dried anti-aging mask containing recombinant human albumin, which contains the following components in parts by mass: the active ingredients are shown in Table 7.

[0091] Table 7 Composition table of various freeze-dried anti-aging masks containing recombinant human albumin

[0092]

[0093] The specific steps for preparing a freeze-dried anti-aging mask containing recombinant human albumin are as follows:

[0094] (1) The grape wine lees polysaccharide extract is prepared by the following method: subject the grape wine lees to high-pressure sterilization (121 °C, 15-30 min), take the sterilized grape wine lees for drying, crush the dried grape wine lees after drying, pass through a 10-mesh sieve to obtain powder, extract with deionized water, soak for 1 h according to a solid-liquid ratio of 1:5, reflux and extract for 15 min at a water bath temperature, extract 3 times, combine the filtrates and concentrate, add 3 times the amount of 95% ethanol to the concentrated solution and place it in the refrigerator for 24 h, centrifuge the polysaccharide precipitate obtained by alcohol precipitation at 5000-8000 r / min for 15-30 min, take the precipitate and suspend it with purified water, and spray-dry the suspended solution to obtain the product.

[0095] (2)The raspberry extract is prepared by the following method: The dried raspberries are crushed and sieved through a 10-mesh sieve to obtain powder, which is soaked in 75% - 80% (v / v) ethanol for 6 - 8 h, the ethanol is distilled off, and then it is suspended in deionized water and spray-dried to obtain the spray-dried powder of the raspberry extract;

[0096] (3)Inlet air temperature for spray drying: 120 - 160 °C, preferably 150 °C; atomization pressure: 0.1 - 1.0 Mpa, preferably 0.5 Mpa; feeding rate: 200 - 800 ml / h, preferably 450 ml / h.

[0097] (4)Take an appropriate amount of deionized water, and successively add the humectant, stabilizer, and thickener into a beaker, stir with a digital constant temperature magnetic stirrer until it is uniform without insoluble substances, then heat to about 80 °C and keep for 20 min, and then cool to below 40 °C as phase A. Take the emollient and stir at 35 - 80 rpm, at a temperature of 55 °C - 85 °C for 15 - 45 min to obtain phase B. Add phase B, phase A, the spray-dried powder of the polysaccharide extract from wine distillers' grains and the spray-dried powder of the raspberry extract in parts by mass to the recombinant human albumin in parts by mass, and use a high-pressure homogenizer at 500 - 1500 bar for high-pressure homogenization for 30 min at a temperature of 25 °C - 50 °C to obtain the mask solution.

[0098] (5)Preparation of the anti-aging freeze-dried mask: The mask solution is evenly dispersed on a hydrogel film, and at the same time, it is freeze-dried using the freeze-drying flash release technology to obtain it. The freeze-drying procedure is that under the pre-freezing preliminary conditions at a temperature of -196 °C, the vacuum degree is 195 - 205 Pa, and sequential addition type sublimation is carried out 2 - 4 times successively.

[0099] Test Example 4: Tyrosinase activity inhibition test of the freeze-dried mask

[0100] Replace the experimental steps in Test Example 1 with the supernatant of each group of masks prepared in Example 2, and the results are shown in Table 8.

[0101] Table 8 Experimental results of tyrosinase activity inhibition test

[0102]

[0103] Note: Compared with Group 8, P ≤ 0.05, * indicates significant difference.

[0104] From the data in Table 8, it can be seen that the inhibition rate of tyrosinase activity of the freeze-dried masks in each group is > 93%. Compared with the results in Table 3, the inhibition rate of tyrosinase activity of the freeze-dried mask in Example 2 is slightly higher than that of the anti-aging composition in Example 1, indicating that the recombinant human albumin anti-aging freeze-dried mask solution prepared by this method has a significant inhibitory effect on tyrosinase and can play an anti-aging role.

[0105] Test Example 5: Determination Experiment on Hygienic Indexes of Freeze-dried Mask and Heavy Metal Determination

[0106] According to the microbial inspection method in the "Technical Specifications for Cosmetics Safety" (2015 Edition), the total number of colonies, thermotolerant Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, molds, and yeasts in a recombinant human albumin mask liquid (Groups 1 - 7) prepared in Example 2 were detected and heavy metals were determined. The microbiological index data are shown in Table 9, and the experimental results of heavy metal detection are shown in Table 10.

[0107] Table 9 Determination Results of Microbiological Indexes for Each Group

[0108]

[0109] Table 10 Determination Results of Heavy Metal Indexes for Each Group

[0110]

[0111] As can be seen from the data in Table 9 and Table 10, the test results of each index in the hygiene of the recombinant human albumin anti-aging freeze-dried mask all meet the regulations.

[0112] Test Example 6: Human Skin Irritation Test of Freeze-dried Mask

[0113] Replace the experimental steps in Test Example 2 with the supernatant of the freeze-dried mask in Groups 1, 4, and 7 prepared in Example 2.

[0114] The experimental results show that the patch test on the skin of the subjects shows that the patch test results of all 30 subjects are negative, indicating that the cream prepared by the invention patent has no obvious irritation and sensitization to the human body and has good safety performance.

[0115] Test Example 7: Anti-wrinkle and Firming Experiment of Freeze-dried Mask

[0116] Replace the experimental steps in Test Example 3 with the supernatant of the mask in each group prepared in Example 2, and the results are shown in Table 11 and Table 12.

[0117] Table 11 Test Results of Elastin

[0118]

[0119] Note: Compared with week 0, P ≤ 0.05, * indicates significant difference.

[0120] According to the indicators of the CBS-805 professional skin tester, the elastin value of the subjects was detected. The higher the value, the firmer the skin. The detected value of this instrument is in the range of 59-70, and the skin state is firm. According to the test results in Table 11: In terms of the elastin index, the content of elastin in the subjects of usage groups 1, 4, and 7 increased significantly after 2 weeks, 4 weeks, and 8 weeks of testing. Among them, the elastin content of the subjects in usage group 4 after 2 weeks, 4 weeks, and 8 weeks of testing was significantly higher than that of the subjects in usage groups 1 and 7, indicating that at different time points, group 4 has a more significant effect on increasing the elastin value. At the same time, the elastin content after 8 weeks of using this product was significantly different compared with that at 0 week. At the same time, compared with the mask without additives, the elastin content was also significantly increased, indicating that the freeze-dried mask prepared by this method can increase the elastin content.

[0121] Table 12 Collagen Test Results

[0122]

[0123] Note: Compared with 0 week, P≤0.05, * indicates significant difference.

[0124] According to the indicators of the CBS-805 professional skin tester, when the detected value is in the range of 25-50, collagen fibers are missing; when it is 50-65, collagen fibers are sparse; when it is greater than 65, collagen fibers are normal. According to the test results in Table 12: In terms of the collagen index, the content of elastin in the subjects of usage groups 1, 4, and 7 increased significantly after 2 weeks, 4 weeks, and 8 weeks of testing. Among them, the collagen content of the subjects in usage group 4 after 2 weeks, 4 weeks, and 8 weeks of testing was significantly higher than that of the subjects in usage groups 1 and 7, indicating that the combination method adopted by group 4 has a more prominent effect on increasing the collagen content. At the same time, the elastin content after using this product for 8 weeks was significantly different compared with that at 0 week. At the same time, when this product was compared with the mask without additives, the elastin content of the users of this product was significantly increased. The mask prepared by this method can effectively increase the elastin content of the skin, thereby improving the skin firmness.

[0125] In summary, the anti-aging freeze-dried mask prepared by this method can significantly promote the regeneration of collagen and elastin, increase the content of collagen and elastin, and has the effect of anti-wrinkle and firming.

[0126] As mentioned above, it is only the embodiments of the present invention. The protection scope of the present invention is not limited by these specific embodiments, but is determined by the claims of the present invention. For those skilled in the art, various changes and modifications can be made to the present invention. Any modification, equivalent replacement, improvement, etc. made within the technical idea and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An anti-aging composition, characterized in that: The composition comprises: The quantity of recombinant human albumin is 0.1~15.0; 0.1-10.0 parts of wine lees polysaccharide extract; 0.1~5.0 parts of red raspberry extract; The preparation steps of the wine lees polysaccharide extract are as follows: sterilizing the wine lees, drying, crushing, and passing through a 10-mesh sieve to obtain a crude extract powder of wine lees polysaccharide, soaking the crude extract with deionized water at a mass ratio of 1:3 to 7 for extraction, concentrating the extract, and then using an ethanol solution with a concentration of 85% v / v or more to precipitate, centrifuging, adding purified water to suspend, and then spray drying to obtain the wine lees polysaccharide extract; The preparation steps of the red raspberry extract are as follows: crushing the dried red raspberries, passing through a 10-mesh sieve to obtain powder, soaking in 75% to 80% v / v ethanol for 6 to 8 hours, distilling to remove the ethanol, then suspending with deionized water, spray drying, and obtaining the red raspberry extract spray-dried powder.

2. The composition according to claim 1, characterized in that The composition may further include additives acceptable in the cosmetic field.

3. The composition according to claim 2, characterized in that The additives include at least one of a preservative, a moisturizer, a thickener, a pH adjuster, a surfactant, a stabilizer, an antioxidant, and a moisturizer.

4. The composition according to any one of claims 1 to 3, characterized in that The dosage form of the composition includes at least one of aqueous solution, emulsion, paste, gel, mask and powder.

5. Use of the composition according to any one of claims 1 to 3 in any of the following aspects: A1: Application in the preparation of anti-aging products; A2: Application in the preparation of products that inhibit tyrosinase activity.

6. Use of the composition according to any one of claims 1 to 3 in the preparation of products promoting the regeneration of collagen and / or elastin.

7. Use of the composition according to any one of claims 1 to 3 in the preparation of firming and anti-wrinkle products.

8. The use according to claim 5, characterized in that: The anti-aging products or products that inhibit tyrosinase activity include cosmetics.

9. The use according to claim 6, characterized in that: The products that promote the regeneration of collagen and / or elastin include cosmetics.

10. The use according to claim 7, characterized in that: The firming and anti-wrinkle products include cosmetics.

11. An anti-aging freeze-dried facial mask, characterized in that: The freeze-dried facial mask comprises: The composition of claim 1; The mass fraction of moisturizer is 5.0~10.0 parts; The mass fraction of thickener is 0.1~0.8 parts; The mass fraction of the moisturizer is 0.3~1.0 parts; The mass fraction of the stabilizer is 0.01~0.20 parts.

12. A freeze-dried facial mask, characterized in that: The freeze-dried facial mask comprises: The composition of claim 1; The mass fraction of glycerol is 5.0~10.0 parts; The mass fraction of carbomer is 0.1~0.8 parts; The mass fraction of ethylhexyl palmitate is 0.3~1.0 parts; The mass fraction of polysorbate 80 is 0.01 to 0.20 parts.

13. A method for preparing an anti-aging composition, characterized in that: The method comprises the following steps: S1: Preparation of wine lees polysaccharide extract: sterilize the wine lees, dry, crush, and pass through a 10-mesh sieve to obtain a crude extract powder of wine lees polysaccharide, soak the crude extract in deionized water at a mass ratio of 1:3 to 7, extract, concentrate the extract, and then precipitate it with an ethanol solution with a concentration of 85% v / v or more, centrifuge, suspend it in purified water, and then spray dry it to obtain the wine lees polysaccharide extract; S2: Preparation of red raspberry extract: crush the dried red raspberries, pass through a 10-mesh sieve to obtain powder, soak them in 75%-80% v / v ethanol for 6-8 hours, remove the ethanol by distillation, suspend them in deionized water, spray dry them, and obtain spray-dried powder of red raspberry extract; S3: Add 0.1-10.0 parts by mass of spray-dried powder of wine sump polysaccharide extract and 0.1-5.0 parts by mass of spray-dried powder of red raspberry extract to 0.1-15.0 parts by mass of recombinant human albumin aqueous solution or salt buffer, and use a high-pressure homogenizer at 500-1000 bar for 5-15 minutes at a temperature of 35°C-60°C to obtain a mixed solution of an anti-aging composition.

14. The method according to claim 13, characterized in that The spray drying conditions in steps S1 and S2 are: air inlet temperature: 120-160° C.; atomization pressure: 0.1-1.0 MPa; feed rate: 200-800 mL / h.

15. The method according to claim 13, characterized in that The method further comprises the step of adding additives acceptable in the field of cosmetics to the anti-aging composition mixed solution obtained in step S3 and mixing the mixture.

Images