Composition containing hydrolase recombinant protein and application thereof

Abstract

The invention discloses a composition containing hydrolase recombinant protein and application of the composition, and particularly belongs to the field of preparations for dressing and makeup. The composition comprises the following components: composite peptide, composite collagen and hydrolase recombinant protein, the composite peptide is composed of palmitoyl tetrapeptide-3, hexapeptide-1, hexapeptide-10, pentapeptide-4 and acetyl hexapeptide-8, and is characterized in that the hexapeptide-3, the hexapeptide-1, the hexapeptide-10, the pentapeptide-4 and the acetyl The composite collagen consists of a recombinant collagen II type, a recombinant collagen VII type and a recombinant collagen X VII type; the hydrolase recombinant protein is composed of recombinant collagenase I type and recombinant hyaluronidase. The composition can effectively promote synthesis of type I collagen, has a promoting effect on proliferation and migration of keratinocytes, and has certain anti-wrinkle and repairing effects; the resident skin external preparation provided by the invention has excellent tightening and anti-wrinkle effects, and meanwhile, the tightening and anti-wrinkle performance has positive correlation with the additive amount of the composition.

Description

Technical Field

[0001] The invention belongs to the field of cosmetic preparations, and in particular relates to a composition containing hydrolase recombinant protein and application thereof in the preparation of anti-wrinkle and repairing preparations. Background Art

[0002] With the growing demand for beauty and skin care, anti-wrinkle and repair functions have become an indispensable part of the cosmetics and skin care market. As the largest organ in the human body, the health and beauty of the skin directly affect the overall image and self-confidence of the individual. However, with the increase of age, environmental factors (such as ultraviolet radiation, air pollution) and bad living habits (such as staying up late, unbalanced diet), the skin gradually shows signs of aging, the most notable of which is the decrease in skin elasticity, the formation of fine lines and wrinkles, and the damage of the skin barrier function.

[0003] During the aging process of the skin, collagen and elastin, as key components for maintaining skin elasticity and firmness, gradually decrease in content with age, causing the skin to lose its original support and form wrinkles. At the same time, the decline in skin cell function also reduces the skin's ability to repair itself, making it difficult to effectively respond to external environmental damage, further exacerbating the problem of skin aging.

[0004] In response to the above problems, a large number of cosmetics and skin care products claiming to have anti-wrinkle and repair functions have emerged on the market. Most of these products add specific active ingredients, such as collagen, hyaluronic acid, vitamin C, peptides, etc., in order to improve skin condition and delay aging. However, due to the existence of the skin barrier and the limitation of skin absorption capacity, a single ingredient is often difficult to fully exert its effect, and may cause skin irritation or allergic reactions due to excessive concentration.

[0005] Therefore, how to develop an anti-wrinkle and repair composition that can integrate the advantages of multiple active ingredients, achieve synergistic effects, and take into account safety and economy has become a technical problem that needs to be urgently solved in the cosmetics and skin care industries. Summary of the invention

[0006] In view of the deficiencies in the prior art, the object of the present invention is to provide a composition with excellent firming, anti-wrinkle and repairing properties and its application in anti-wrinkle and repairing preparations. The components in the composition synergize with each other and can effectively promote collagen synthesis and skin barrier repair.

[0007] In order to achieve the above object, the present invention discloses the following technical solutions:

[0008] In a first aspect, the present invention provides a composition containing a hydrolase recombinant protein, wherein the composition comprises the following components by weight:

[0009] Composite peptide 4.1-4.6 parts;

[0010] Composite collagen 2.8-3.2 parts;

[0011] Hydrolase recombinant protein 0.007-0.009 parts;

[0012] The composite peptide consists of palmitoyl tetrapeptide-3, hexapeptide-1, hexapeptide-10, pentapeptide-4, and acetyl hexapeptide-8;

[0013] The composite collagen is composed of recombinant collagen type II, recombinant collagen type VII and recombinant collagen type XVII;

[0014] The hydrolase recombinant protein consists of recombinant collagenase type I and recombinant hyaluronidase.

[0015] Preferably, the mass ratio of palmitoyl tetrapeptide-3, hexapeptide-1, hexapeptide-10, pentapeptide-4 and acetyl hexapeptide-8 is (4.1-4.5):(0.2-0.5):(1.2-1.4):(2.5-3.1):1.

[0016] Preferably, the mass ratio of the recombinant collagen type II, recombinant collagen type VII and recombinant collagen type XVII is 1:(1.3-2.0):(0.7-1.0).

[0017] Preferably, the mass ratio of the recombinant collagenase type I to the recombinant hyaluronidase is 1:(0.5-1.0).

[0018] In a second aspect, the present invention provides use of the composition containing the hydrolase recombinant protein described in the first aspect in the preparation of a skin care preparation having anti-wrinkle and / or repairing effects.

[0019] In a third aspect, the present invention provides an anti-wrinkle and repairing resident skin external preparation, comprising the composition containing the hydrolase recombinant protein according to the first aspect;

[0020] The amount of the composition added to a leave-on skin external preparation is ≤0.5 wt %.

[0021] Preferably, moisturizers, emollients, preservatives, thickeners, antioxidants, pH regulators and solvents are also included.

[0022] Further preferably, the moisturizer is at least one of butylene glycol, 1,2-hexanediol, glycerin, sodium hyaluronate, and panthenol;

[0023] The emollient is at least one of C10-18 fatty acid triglycerides, coconut oil alcohol-caprylate / caprate, and tridecyl trimellitate;

[0024] The thickener is at least one of xanthan gum and cellulose gum;

[0025] The preservative is p-hydroxyacetophenone;

[0026] The antioxidant is at least one of niacinamide and vitamin C;

[0027] The pH regulator is arginine;

[0028] The solvent is water.

[0029] In a fourth aspect, the present invention provides a method for preparing the anti-wrinkle and repairing residual skin external preparation according to the third aspect, characterized in that it comprises the following steps:

[0030] S1, add each component of component A into a reaction container, stir and heat to 80-85°C to dissolve and mix evenly, keep warm for 15 minutes, and start cooling to 45°C to obtain mixture A;

[0031] S2, heating each component of component B to 80°C until completely dissolved, maintaining 80°C for mixing and stirring evenly, then cooling to 45°C for standby use;

[0032] S3, adding the component B dissolved and stirred evenly in step S2 to the mixture A, and adding the components C and D in sequence, maintaining the temperature at 45° C. and stirring and dispersing evenly, cooling to 35° C. and adding a pH adjuster to adjust the pH value to 6.0-7.0, and filtering to obtain a leave-on skin topical preparation;

[0033] The components of component A include xanthan gum, cellulose gum, glycerin, sodium hyaluronate and water;

[0034] The B component includes 1,2-hexanediol, p-hydroxyacetophenone, C10-18 fatty acid triglycerides, coconut oil alcohol-caprylate / caprate, tridecyl trimellitate and butylene glycol;

[0035] The C component includes panthenol, vitamin C, niacinamide and water;

[0036] The D component includes a composition containing a hydrolase recombinant protein and water.

[0037] Preferably, the step S1 is stirred at 120 r / min;

[0038] In step S2, the mixture is mixed and stirred at 30 r / min and then cooled;

[0039] In the step S3, the mixture is stirred and dispersed uniformly at 200 r / min and then cooled to 35° C., and the filtration is performed through a 100-mesh filter cloth.

[0040] In the present invention:

[0041] The composite peptide provided by the present invention comprises a mixture of multiple different peptide compounds including palmitoyl tetrapeptide-3, hexapeptide-1, hexapeptide-10, pentapeptide-4 and acetyl hexapeptide-8. These peptide compounds have different biological activities, such as anti-aging, anti-oxidation, wrinkle removal and whitening. They can stimulate the production of collagen and elastin, improve skin elasticity and firmness; they can also inhibit muscle contraction and reduce the formation of wrinkles.

[0042] Composite collagen is composed of recombinant collagen type II, recombinant collagen type VII and recombinant collagen type XVII. It is a compound substance that can replenish collagen in the skin by penetrating deeper into the skin, improving the condition of the skin. At the same time, it has a good moisturizing effect, can absorb and retain a large amount of water molecules, and keep the skin hydrated and smooth. In addition, it can also promote the production of collagen and enhance the elasticity and firmness of the skin.

[0043] Recombinant collagenase type I plays a regulating and promoting role in catalyzing the degradation and reconstruction of collagen in the body. Collagenase has the ability to degrade collagen and stimulate the synthesis of new collagen, making the skin firmer and smoother. At the same time, it can break down large-molecule collagen into small-molecule peptides through hydrolysis, thereby increasing the skin's absorption rate of collagen, more effectively replenishing collagen in the skin, improving skin elasticity and reducing wrinkles. Collagenase can work synergistically with other ingredients such as hyaluronic acid, vitamins and plant extracts to enhance the overall effect of cosmetics.

[0044] Recombinant hyaluronidase is a recombinant enzyme protein whose main function is to degrade or break down hyaluronic acid molecules. Hyaluronic acid is a polysaccharide and a component with important functions in human skin tissue. It exists in collagen and elastin, and together with them maintains the elasticity and moisture of the skin, making the skin look smooth and firm. Hyaluronidase can promote the metabolism of hyaluronic acid in the skin, promote skin renewal, and improve the skin's repair ability.

[0045] The present invention finds that when the components of composite peptides, composite collagen and hydrolase recombinant protein are combined in a certain proportion and used in combination, they can promote each other and exert a more excellent firming and anti-wrinkle effect. Therefore, while maintaining the same or better skin care effect, the addition amount of each component can be significantly reduced, thereby reducing the cost of the product and improving safety.

[0046] Beneficial effects of the present invention:

[0047] 1. The composition provided by the present invention can effectively promote the synthesis of type I collagen, and at the same time promote the proliferation and migration of keratinocytes, and has certain anti-wrinkle and repair effects;

[0048] 2. The leave-on skin topical preparation provided by the present invention has excellent firming and anti-wrinkle effects. At the same time, the firming and anti-wrinkle performance has a certain dependence on the additive amount of the composition containing the hydrolase recombinant protein. DETAILED DESCRIPTION

[0049] The technical solutions in the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0050] Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in the field without making any creative work shall fall within the scope of protection of the present invention.

[0051] The technical solution of the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only used to explain the present invention and are not used to limit the present invention.

[0052] In the present invention:

[0053] raw material Purchased from Palmitoyl Tetrapeptide-3 Shanghai Qiangyao Biotechnology Co., Ltd. Hexapeptide-1 Guangzhou Novo Biotechnology Co., Ltd. Hexapeptide-10 Shanghai Qiangyao Biotechnology Co., Ltd. Pentapeptide-4 Shanghai Qiangyao Biotechnology Co., Ltd. Acetyl Hexapeptide-8 Guangzhou Novo Biotechnology Co., Ltd. Recombinant collagen type II Jiangsu Jiangshan Juyuan Biotechnology Co., Ltd. Recombinant collagen type VII Jiangsu Jiangshan Juyuan Biotechnology Co., Ltd. Recombinant collagen type XVII Jiangsu Jiangshan Juyuan Biotechnology Co., Ltd. Recombinant collagenase type I Beijing Biolab Technology Co., Ltd. Recombinant Hyaluronidase Beijing Biolab Technology Co., Ltd. Other raw materials Commercially available

[0054] Preparation of composite peptides:

[0055] The composite peptide was obtained by compounding the raw materials according to the mass ratio in Table 1.

[0056] Table 1. Mass ratio of composite peptide raw materials

[0057] raw material Composite peptide 1 Composite peptide 2 Composite peptide 3 Composite peptide 4 Composite peptide 5 Composite peptide 6 Composite peptide 7 Composite peptide 8 Palmitoyl Tetrapeptide-3 4.1 4.3 4.5 1 - 4.1 4.1 4.1 Hexapeptide-1 0.5 0.3 0.2 1 0.5 - 0.5 0.5 Hexapeptide-10 1.2 1.3 1.4 1 1.2 1.2 - 1.2 Pentapeptide-4 2.5 2.8 3.1 1 2.5 2.5 2.5 - Acetyl Hexapeptide-8 1 1 1 1 1 1 1 1 raw material Composite peptide 9 Compound peptide 10 Composite peptide 11 Compound peptide 12 Composite peptide 13 / / / Palmitoyl Tetrapeptide-3 4.1 4.1 4.1 4.1 4.1 / / / Hexapeptide-1 0.5 0.5 - - - / / / Hexapeptide-10 1.2 - 1.2 - - / / / Pentapeptide-4 2.5 - - 2.5 - / / / Acetyl Hexapeptide-8 - - - - 1 / / /

[0058] Note: “-” in the table means not added.

[0059] Composite peptide performance test:

[0060] 1. Test consumables:

[0061] The composite peptide prepared above and deionized water were prepared into test samples at a concentration of 1 wt%, with a total of 13 groups;

[0062] Healthy 10-week-old nude mice, male, weighing about 20 g;

[0063] Skin hydroxyproline assay kit;

[0064] Spectrophotometer.

[0065] 2. Test methods:

[0066] The test samples were applied to the right ear of nude mice, once in the morning and evening, with a dosage of 1g. The left side was not treated as a blank control group. The mice were fed with the same feed in a clean, ventilated, dry environment at room temperature of 22±2℃. Five mice were taken from each group 30 days after the medication, and the skin at the drug action site and the corresponding site on the opposite side was taken.

[0067] Weigh 45 mg of the tissue wet weight on a balance and put it into a clean test tube. Add 1 mL of the hydrolyzate in the hydroxyproline assay kit and mix well. Cover and hydrolyze in a boiling water bath for 20 minutes, mixing every 10 minutes to make the hydrolysis more complete. Take out the test tube, cool it, add 1 drop of the indicator in the kit to each tube, and shake it well. Add 1 mL of solution A in the kit to adjust the pH and shake it well. Then use a pipette to absorb the pH-adjusting solution B and add it drop by drop to each test tube until the indicator in each tube turns yellow-green. Add distilled water to 10 mL and mix well. Take 3 mL of the hydrolyzate, add 20-30 mg of activated carbon and mix well. Centrifuge at 3500 rpm for 10 minutes and take 1 mL of the supernatant.

[0068] Take a clean test tube and add 1mL of 5μg / mL standard application solution as the standard tube. Add reagent 1 to each tube and mix well, let stand for 10 minutes, then add reagent 2, mix well and let stand for 5 minutes, add reagent 3, mix well in a 60℃ water bath for 15 minutes, cool and centrifuge at 3500 rpm for 10 minutes, take the supernatant and measure the absorbance at 550nm, 1cm optical path (specific standard application solution, reagents 1, 2, 3 are prepared according to the instructions of the detection kit).

[0069] 3. Result determination and analysis

[0070] Hydroxyproline is a unique amino acid in collagen, and the content of hydroxyproline is often used to convert the content of collagen. The content of hydroxyproline (HYP) was determined by spectrophotometry, and the content of collagen in mouse skin was converted according to 14% HYP in collagen.

[0071] Table 2 Effects of each group of test samples on collagen production in mouse skin

[0072] Grouping Collagen content in skin (%) (30 days) Blank control group 13.21 Composite peptide 1 37.48** Composite peptide 2 41.22** Composite peptide 3 39.71** Composite peptide 4 29.73# Composite peptide 5 21.11# Composite peptide 6 23.45# Composite peptide 7 21.52# Composite peptide 8 17.33## Composite peptide 9 20.06## Compound peptide 10 22.17# Composite peptide 11 24.92# Compound peptide 12 23.71# Composite peptide 13 25.16#

[0073] Note: ** indicates that the difference between each composite peptide 1-3 and the blank control group is extremely significant, p < 0.01. # indicates that the difference between composite peptide 1 and composite peptide 4-13 is significant, p < 0.05, ## indicates that the difference is extremely significant, p < 0.01.

[0074] Result analysis:

[0075] According to the test results in Table 2, palmitoyl tetrapeptide-3, hexapeptide-1, hexapeptide-10, pentapeptide-4, and acetyl hexapeptide-8 in the composite peptide can promote high expression of collagen under a certain mass ratio. At the same time, the components in the composite peptide are indispensable, and the lack of related components will lead to weakened performance; composite peptide 2 has the best performance, and composite peptide 2 is selected as the raw material for subsequent preparation;

[0076] Preparation of composite collagen:

[0077] The composite collagen was obtained by compounding the raw materials according to the mass ratio of Table 3.

[0078] Table 3. Mass ratio of composite collagen raw materials

[0079]

[0080] Note: “-” in the table means not added.

[0081] Preparation of hydrolase recombinant protein:

[0082] The hydrolase recombinant protein was obtained by compounding the raw materials according to the mass ratio of Table 4.

[0083]

[0084] Note: “-” in the table means not added.

[0085] Preparation of the composition:

[0086] The raw materials were compounded according to the mass fractions in Table 5 to obtain the composition.

[0087] Table 5 Composition raw materials weight parts

[0088]

[0089] Note: “-” in the table means not added.

[0090] Composition performance test:

[0091] 1. Test to promote the synthesis of type I collagen

[0092] In this experiment, fibroblasts were used as the test object, and a cell photoaging damage model was established through UVA irradiation. The changes in the content of type I collagen (Collagen I) after the test sample was exposed to the test sample were detected to evaluate the firming and anti-wrinkle effects of the test sample.

[0093] (1) Inoculation: 1×10 4 The cells were inoculated into a 96-well plate at a density of 100 cells / well and placed in an incubator for overnight incubation (37° C., 5% CO 2 ).

[0094] (2) UVA irradiation: After washing the cells with PBS, the UVA irradiation group was irradiated with 30 J / cm 2 of UVA radiation.

[0095] (2) Solution preparation: Dissolve compositions 1-12 in deionized water to prepare a composition solution with a concentration of 0.5 wt%, which is the sample solution to be tested; TGF-β (100 ng / mL) is used as a positive control for the experiment. Add 100 uL to each well, and set up 3 replicate wells for each group. After the administration is completed, place the 96-well plate in a 37°C incubator away from light and incubate for 24 hours.

[0096] (4) Collection of cell supernatant: 24 h after administration, the 96-well plate was removed and the cell supernatant corresponding to each sample group was collected into a centrifuge tube. The cells were centrifuged at 1000 rpm for 10 min. The supernatant was collected and placed in a 1.5 mL centrifuge tube and stored at -20°C for later use.

[0097] (5) Determination of type I collagen concentration in cell supernatant using ELISA kit: Take out the kit and the sample to be tested 30 min before the test and place them at room temperature before use. The test operation shall be performed strictly according to the instructions of the kit.

[0098] Table 6 Type I collagen content detection results

[0099]

[0100]

[0101] 2. Effect of the composition on repairing the skin barrier

[0102] 1. Experimental Materials

[0103] 1.1 Experimental cells

[0104] Human immortalized keratinocytes (HaCaT).

[0105] 1.2 Samples and reagents

[0106] Composition 1-12, fetal bovine serum (FBS), DMEM culture medium, dimethyl sulfoxide (DMSO).

[0107] 2. Experimental Methods

[0108] 2.1 Detection of the effect of promoting skin keratinocyte scratch healing

[0109] 2.1.1 HaCaT cell lines were cultured in a 37°C cell culture incubator using DMEM containing 10% FBS, with the culture medium replaced every 24 hours. Cells were passaged to 80% and maintained in logarithmic growth. After the cells reached the third generation and were stable, they were used in the experiment.

[0110] 2.1.2 Take 10 mg of composition 1-12 and 100 μL DMSO respectively, vortex for 2-3 min to mix evenly, add 900 μL DMEM culture medium, vortex to mix evenly to prepare the stock solutions (10 mg / mL) of each group. The culture medium in the cell experiment was prepared by diluting the stock solutions (10 mg / mL) of each group in proportion.

[0111] 2.1.3 HaCaT cells in the logarithmic growth phase were digested and made into a cell suspension using DMEM complete medium containing 10% fetal bovine serum. HaCaT cells were plated at 4×10 4 Each well was inoculated into the cell insert and cultured in a constant temperature incubator at 37°C and 5% CO2.

[0112] 2.1.4 When the cell density is about 80%, remove the cell plug and the liquid in the 6-well plate, and divide the experiment into a blank control group and combination groups 1 to 12. Dilute the stock solution (10 mg / mL) of each group in 2.1.2 with DMEM medium to an intervention concentration of 20 μg / mL, add 2 mL / well respectively, and continue to culture in an incubator.

[0113] 2.1.5 Take photos at 0h and 24h, calculate the cell area and the corresponding migration rate using ImageJ. Migration rate = (0h area - 24h area) / 0h area, relative migration rate = sample migration rate - blank group migration rate, the results are shown in Table 7.

[0114] Table 7 Effect of the composition on the migration rate of skin keratinocytes

[0115]

[0116]

[0117] Preparation of leave-on skin topical preparations

[0118] The specific raw materials are as shown in Table 8;

[0119] Step S1, add each component of component A into a reaction kettle, stir at 120r / min and heat to 80-85°C to dissolve, mix and stir evenly, keep warm for 15 minutes, and start cooling;

[0120] Step S2, heating each component of component B to 80°C until completely dissolved, maintaining 80°C at 30r / min for mixing and stirring, then cooling to 45°C for standby use;

[0121] Step S3, after the reaction kettle is cooled to 45°C, the B component dissolved and stirred evenly in step S2 is added into the reaction kettle under stirring, and component C and component D are added in sequence, and the temperature is maintained at 45°C and stirred at 200r / min to disperse evenly. After cooling to 35°C, a pH adjuster is added to adjust the pH value to 6.0-7.0, and filtered through a 100-mesh filter cloth to obtain a residual skin topical preparation.

[0122] Table 8 Raw materials for skin external preparations

[0123]

[0124]

[0125] Note: The units in the table are mass percentage.

[0126] Embodiment performance test:

[0127] 1. Test method:

[0128] Fifteen volunteers aged 30±3 years with many fine lines and wrinkles, loose, inelastic and dry faces were selected. The samples were used for 4 weeks. The skin elasticity R2 value and skin elasticity F4 value (average value) of each group of volunteers were measured by MPA580 multifunctional skin tester to evaluate the anti-wrinkle and firming effects.

[0129] Take the skin external preparation examples 1-3 prepared above as test samples and use them according to the following table.

[0130]

[0131] Before the experiment, volunteers carefully read and signed the informed consent form. The samples were confirmed to be mild to the skin and no adverse reactions occurred through human patch tests. The experiment adopted a before-and-after comparison method. The test period was 4 weeks and the skin data was collected 3 times, namely 0 days, 14 days and 28 days.

[0132] 2. Evaluation basis:

[0133] Skin elasticity R2 value: the total elastic-plasticity of the rebound part / the total elastic-plasticity of the stretching part. The closer the value is to 100%, the greater the elasticity of the curve. It characterizes the viscoelasticity of the skin. The higher the value, the better the skin elasticity.

[0134] Skin elasticity F4 value: It is the area value within the envelope curve formed during the application and elimination of negative pressure in all test cycles. The smaller the value, the tighter the skin.

[0135] 3. Experimental results:

[0136] Table 9 Skin elasticity R2 value test results

[0137]

[0138] Table 10 Skin elasticity F4 value test results

[0139]

[0140]

[0141] Result analysis:

[0142] According to the test results in Tables 6 and 7, compositions 1-3 have a promoting effect on the synthesis of type I collagen, indicating that the compositions provided by the present invention have a certain firming and anti-wrinkle effect. Compositions 1-3 have the greatest effect on the migration rate of skin keratinocytes. The cell scratch test is a simple and quick method for measuring cell migration and repair ability. Similar to the in vitro wound healing model, the detection of cell migration rate can reflect the barrier repair ability of cells. The greater the migration rate, the stronger the barrier repair ability, indicating that the compositions provided by the present invention have a certain repair effect on the skin; in the performance test, the components in the composition were reduced accordingly, resulting in a certain degree of weakening of the performance of the composition, indicating that the components in the composition are indispensable and the components are interdependent under a certain ratio;

[0143] According to the test results in Tables 9 and 10, the skin external preparation provided by the present invention has a firming and anti-wrinkle effect. At the same time, the firming and anti-wrinkle effect has a certain dependence on the concentration of the composition.

[0144] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A composition containing a hydrolase recombinant protein, characterized in that: The composition comprises the following components by weight: Composite peptide 4.1-4.6 parts; Composite collagen 2.8-3.2 parts; Hydrolase recombinant protein 0.007-0.009 parts; The composite peptide consists of palmitoyl tetrapeptide-3, hexapeptide-1, hexapeptide-10, pentapeptide-4, and acetyl hexapeptide-8; The composite collagen is composed of recombinant collagen type II, recombinant collagen type VII and recombinant collagen type XVII; The hydrolase recombinant protein consists of recombinant collagenase type I and recombinant hyaluronidase.

2. The composition containing hydrolase recombinant protein according to claim 1, characterized in that: The mass ratio of palmitoyl tetrapeptide-3, hexapeptide-1, hexapeptide-10, pentapeptide-4 and acetyl hexapeptide-8 is (4.1-4.5):(0.2-0.5):(1.2-1.4):(2.5-3.1):

1.

3. The composition containing hydrolase recombinant protein according to claim 1, characterized in that: The mass ratio of the recombinant collagen type II, recombinant collagen type VII and recombinant collagen type XVII is 1:(1.3-2.0):(0.7-1.0).

4. The composition containing hydrolase recombinant protein according to claim 1, characterized in that: The mass ratio of the recombinant collagenase type I to the recombinant hyaluronidase is 1:(0.5-1.0).

5. Use of the composition containing the hydrolase recombinant protein according to any one of claims 1 to 4 in the preparation of a skin care preparation with anti-wrinkle and / or repairing effects.

6. An anti-wrinkle and repairing leave-on skin external preparation, characterized in that: A composition comprising a hydrolase recombinant protein according to any one of claims 1 to 4; The amount of the composition added to a leave-on skin external preparation is ≤0.5 wt %.

7. The anti-wrinkle and repairing leave-on skin external preparation according to claim 6, characterized in that: It also includes moisturizers, emollients, preservatives, thickeners, antioxidants, pH adjusters, and solvents.

8. The anti-wrinkle and repairing leave-on skin external preparation according to claim 7, characterized in that: The moisturizing agent is at least one of butylene glycol, 1,2-hexanediol, glycerin, sodium hyaluronate, and panthenol; The emollient is at least one of C10-18 fatty acid triglycerides, coconut oil alcohol-caprylate / caprate, and tridecyl trimellitate; The thickener is at least one of xanthan gum and cellulose gum; The preservative is p-hydroxyacetophenone; The antioxidant is at least one of niacinamide and vitamin C; The pH regulator is arginine; The solvent is water.

9. A method for preparing the anti-wrinkle and repairing leave-on skin external preparation according to claim 7, characterized in that: The following steps are involved: S1, add each component of component A into a reaction container, stir and heat to 80-85°C to dissolve and mix evenly, keep warm for 15 minutes, and start cooling to 45°C to obtain mixture A; S2, heating each component of component B to 80°C until completely dissolved, maintaining 80°C for mixing and stirring, then cooling to 45°C for standby use; S3, adding the component B dissolved and stirred evenly in step S2 to the mixture A, and adding the components C and D in sequence, maintaining the temperature at 45° C. and stirring and dispersing evenly, cooling to 35° C. and adding a pH adjuster to adjust the pH value to 6.0-7.0, and filtering to obtain a leave-on skin topical preparation; The components of component A include xanthan gum, cellulose gum, glycerin, sodium hyaluronate and water; The B component includes 1,2-hexanediol, p-hydroxyacetophenone, C10-18 fatty acid triglycerides, coconut oil alcohol-caprylate / caprate, tridecyl trimellitate and butylene glycol; The C component includes panthenol, vitamin C, niacinamide and water; The D component includes a composition containing a hydrolase recombinant protein and water.

10. The method for preparing the anti-wrinkle and repairing leave-on skin external preparation according to claim 9, characterized in that: The step S1 is stirring at 120 r / min; In step S2, the mixture is mixed and stirred at 30 r / min and then cooled; In the step S3, the mixture is stirred and dispersed uniformly at 200 r / min and then cooled to 35° C., and the filtration is performed through a 100-mesh filter cloth.

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