CN110772446B

CN110772446B - Skin repair polypeptide composition of water-in-oil system

Info

Publication number: CN110772446B
Application number: CN201911319241.1A
Authority: CN
Inventors: 丁文锋; 观富宜; 彭晏; 黄春青
Original assignee: Shenzhen Weiqi Technology Co ltd
Current assignee: Shenzhen Weiqi Technology Co ltd
Priority date: 2019-12-19
Filing date: 2019-12-19
Publication date: 2023-08-22
Anticipated expiration: 2039-12-19
Also published as: CN110772446A

Abstract

The invention discloses a skin repair polypeptide composition of a water-in-oil system, which belongs to the technical field of cosmetic polypeptides, and comprises polypeptides with collagen generation promoting effect, wherein the polypeptides exist in a water phase, the mass percentage concentration of each polypeptide is 0.0001% -5%, and the water phase and an oil phase form a water-in-oil system. The oil phase comprises a surfactant, an oily emollient. Compared with a water-based system, the skin repair polypeptide composition adopts a water-in-oil system, and polypeptide components are dissolved in a small amount of water phase, so that the local polypeptide concentration in the water-in-oil system is higher, the skin repair polypeptide composition is easier to be absorbed through skin, the accumulation amount of the skin is higher, the same feeding amount can achieve a better skin repair effect, and the skin repair polypeptide composition has the advantages of good stability, high safety, simple preparation process and low cost, and is suitable for industrial mass production.

Description

Skin repair polypeptide composition of water-in-oil system

Technical Field

The invention belongs to the technical field of cosmetic polypeptides, and relates to a water-in-oil system skin repair polypeptide composition.

Background

In human skin, 70% are composed of collagen. With the increase of age and the influence of ultraviolet rays, collagen is lost in a large amount, collagen synthesized by fibroblasts and glycosaminoglycan are reduced, so that collagen peptide bonds and elastic nets supporting skin are hardened and broken, the reticular structure of dermis of the skin is loose and then destroyed, collapse occurs, and the skin is loose, wrinkled and dry.

In the natural skin aging defending and nursing process, the active polypeptide plays a unique and important physiological role, is taken as an important molecule in the natural skin repairing process, participates in the skin repairing process in a plurality of natural states, not only can promote the proliferation of skin cells, but also can provide nutrition for the skin, delay the skin aging and promote the skin wound repair, and draws extensive attention of researchers. Polypeptides are currently widely added to skin care products as one of the active ingredients for skin repair. These active polypeptides are all water-soluble, and thus the products on the market are also water-based, in which the polypeptides are dissolved. This presents another problem in that the polypeptide is easily degraded in an aqueous solution, and the addition of the polypeptide component directly to the water-based product results in poor stability of the polypeptide in the product, so that the polypeptide originally having high activity cannot sufficiently exert its intended skin repair effect, and degradation products generated by degradation of the polypeptide in the product may potentially harm human body. In addition, water-soluble substances are difficult to be absorbed through the skin barrier, and thus it is difficult to sufficiently exert their skin-repair effects. In order to increase transdermal absorption to achieve the desired effect, it is necessary to increase the amount of the material to be fed, thereby bringing about an increase in cost.

Therefore, there is an urgent need in the art for a skin repair product that has good stability, high safety, easy transdermal absorption, low cost, simple preparation process, suitability for industrial mass production, excellent effect, and capability of compensating for the defects of the prior art.

Disclosure of Invention

The technical problem to be solved by the invention is to provide the skin repair polypeptide composition which has the advantages of good stability, high safety, easy transdermal absorption, low cost, simple preparation process, suitability for industrial mass production and excellent effect.

Through experimental research, the inventor finds that polypeptide components are dispersed in an oil phase to form a water-in-oil system, a small amount of water phase in the system can reduce the contact between the polypeptide and water, so that the polypeptide is concentrated in the water phase, the stability of the polypeptide is improved, and the polypeptide with larger local concentration is beneficial to exerting better skin repairing effect; the external oil phase can make polypeptide component penetrate the skin barrier better and promote absorption, so that the product has more excellent skin repairing effect, and the invention is formed.

The invention provides a water-in-oil system skin repair polypeptide composition, which comprises polypeptides with collagen generation promoting effect, and is selected from one or more of the following polypeptides, including but not limited to: palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, palmitoyl pentapeptide-4, tripeptide-1, tetrapeptide-21, palmitoyl hexapeptide-6, palmitoyl tripeptide-5, myristoyl pentapeptide-11, myristoyl hexapeptide-4, myristoyl octapeptide-1, hexapeptide-3, tripeptide-1 copper, wherein the polypeptide with collagen production promoting effect is present in an aqueous phase, the mass percentage concentration of each polypeptide is 0.0001% -5%, and the aqueous phase and the oil phase form a water-in-oil system.

The polypeptide composition further comprises a polypeptide having a laminin production promoting effect selected from one or more of the following polypeptides, including but not limited to: hexapeptide-8, hexapeptide-9, hexapeptide-10, palmitoyl dipeptide-5 diaminobutyryl hydroxythreonine and palmitoyl dipeptide-6-diaminohydroxybutyric acid, wherein the mass percentage concentration of each polypeptide is 0.0001-5%.

The polypeptide composition may further comprise a polypeptide having an elastin-promoting effect, including, but not limited to: palmitoyl hexapeptide-12 with polypeptide mass percentage concentration of 0.0001% -5%.

The polypeptide composition may further comprise a polypeptide having a complement to the proteoglycan deficiency, selected from one or more of the following polypeptides, including but not limited to: acetyl tetrapeptide-9, acetyl tetrapeptide-11 and tripeptide-10citrulline, wherein the mass percentage concentration of each polypeptide is 0.0001-5%.

The oil phase of the present invention comprises a surfactant and an oily emollient.

The surfactant is one or a combination of more of PEG-20 triisostearate, PEG-7 glycerol cocoate, PEG-6 caprylic/capric glyceride, polyglycerol-2 isostearate and polyglycerol-4 laurate, and the mass percentage concentration of each component is 1-10%.

The oily emollient is one or a combination of a plurality of hydrogenated polyisobutene, caprylic/capric triglyceride and isononyl isononanoate, and the mass percentage concentration of the oily emollient is 70-90%.

The oily emollient may further comprise a vegetable oil which is camellia seed oil, jojoba seed oil, nut oil, sunflower seed oil.

The water-in-oil system skin repair polypeptide composition is mainly used for preparing skin care products or medical products for external use.

In order to facilitate understanding of the present invention, the principle of the present invention, the mechanism of action of skin repair polypeptides of different action targets is described as follows:

1. principles of the invention

Analysis of skin structure shows that collagen is the major protein component in skin and accounts for about 70% of skin protein content. The collagen can be further crosslinked to form collagen fibers, so that the skin connection strength and stability are improved, and the collagen fibers can support the skin. With age, the collagen content in skin gradually decreases, which causes the skin to lose elasticity, and wrinkles, skin slackening, dryness, etc. appear. Laminin is the protein in the extracellular matrix (ECM) with the greatest content other than collagen, is an indispensable part of skin composition, and is combined with various basement membrane components to regulate cell growth and differentiation. Elastin is the main component of elastic fiber, responsible for maintaining and supporting the elasticity of the skin, giving it its elasticity and flexibility. With age, elastin in the skin gradually runs off and the skin becomes loose. Proteoglycans are also important components of the extracellular matrix (ECM) and can maintain skin integrity, making the skin soft and elastic.

Based on the characteristics of the skin structural components, the skin repair polypeptide composition comprises a polypeptide with the function of promoting collagen production, and the polypeptide with the function of promoting laminin production, the polypeptide with the function of promoting elastin production and the polypeptide with the function of compensating proteoglycan deficiency are added on the basis.

2. Mechanism of action of skin repair polypeptides

2.1 polypeptide action mechanism with collagen production promoting action

Palmitoyl tripeptide-1 (Palmitoyl Tripeptide-1, pal-GHK, CAS number: 147732-56-7) is obtained by modifying the N end of the peptide chain of GHK with palmitic acid, has better transdermal performance, can promote skin to synthesize a large amount of collagen and polysaccharide, and can tighten skin, lock water and moisturize, lighten wrinkles and deeply repair skin.

Palmitoyl tetrapeptide-7 (Palmitoyl Tetrapeptide-7, CAS number 221227-05-0) helps promote collagen production and repair thickened epidermis. In addition, palmitoyl tetrapeptide-7 may regulate the secretion of interleukin-6 (IL-6). IL-6 participates in chronic inflammatory reaction of skin, and the increase of IL-6 accelerates skin aging, and palmitoyl tetrapeptide-7 can reduce the level of IL-6 in the skin aging process, thereby re-maintaining the balance of cytokines in skin, remarkably eliminating skin inflammation and reducing glycosylation damage, so as to achieve the purposes of delaying skin aging and increasing skin elasticity and compactness.

Palmitoyl pentapeptide-4 (Palmitoyl Pentapeptide-4) can stimulate fibroblast proliferation and stimulate synthesis of collagen, glycosaminoglycans (GAGs) and hyaluronic acid, so as to increase skin thickness and achieve the purpose of skin repair.

Tripeptide-1 (GHK), an endogenous growth factor, acts as a transporter for copper ions, stimulating collagen synthesis, angiogenesis and fibroblast growth. The GHK with proper dosage has anti-inflammatory and repairing effects on skin. Furthermore, marc Heidl et al found that tripeptide-1 could promote the synthesis of skin hyaluronic acid. Therefore, the tripeptide-1 can be used for repairing skin, moisturizing and improving skin quality.

Tetrapeptide-21 (gekg) can promote collagen production in extracellular matrix (ECM), and induce increased expression of collagen gene, thereby thickening skin and improving skin roughness.

Palmitoyl hexapeptide-6 (Palmitoyl Hexapeptide-6) is a peptide developed by taking hereditary immune peptide as a template, and can effectively stimulate fibroblast proliferation and linking, promote collagen synthesis, repair dermis and improve skin quality.

Palmitoyl tripeptide-5 (Palmitoyl Tripeptide-5) stimulates fibroblast to divide and proliferate by activating transforming growth factor-beta (TGF-beta), strongly promotes collagen production, supplements extracellular matrix protein, strengthens connective tissue, inhibits the activity of matrix metalloproteinase, prevents collagen from decomposing, and thus makes skin elastic and deeply repairs skin barrier.

Myristoyl pentapeptide-11 (Myristoyl Pentapeptide-11) can help restore skin elasticity, repair damaged skin, stimulate collagen synthesis, thereby increasing skin water storage capacity and achieving the effects of moisturizing. Similar mechanisms of action are myristoyl hexapeptide-4 (Myristoyl Hexapeptide-4) and myristoyl octapeptide-1 (Myristoyl Octapeptide-1).

Hexapeptide-3 (Hexapeptide-3) is a small molecule peptide resembling the 6-amino acid fragment sequence in fibronectin type III units, promoting the expression of β1 integrin, collagen adhesion, enhancing the adhesion of cells to extracellular matrix (ECM), and thus promoting tissue remodeling and skin repair.

Tripeptide-1 Copper (Copper Peptide) can promote the mass production of collagen, elastin, proteoglycan and glycosaminoglycan (GAGs), increase skin elasticity, and tighten skin, thereby smoothing wrinkles; antioxidant, free radical scavenging, and skin aging delaying; promoting proliferation of epidermal cells and repairing skin barrier.

2.2 polypeptide mechanism of action with laminin production promoting action

Hexapeptide-8 (Hexapeptide-8) compacts the skin by stimulating collagen production of laminin-V and type IV, VII. Also similar in mechanism of action are Hexapeptide-10 (Hexapeptide-10), palmitoyl dipeptide-5 diaminobutyryl hydroxythreonine (Palmitoyl Dipeptide-5Diaminobutyroyl Hydroxythreonine) and palmitoyl dipeptide-6Diaminohydroxybutyrate (Palmitoyl Dipeptide-6 diaminobutyrate).

Hexapeptide-9 (Hexapeptide-9) can increase the amount of laminin-V and integrin, increase the synthesis of collagen in dermis, promote the regeneration and reinforcement of structural tissues in Dermis Epidermis Junction (DEJ) area, promote the differentiation of keratinocytes and the synthesis of keratin, thereby repairing wounds, recombining complete epidermis layers, promoting the regeneration of skin, enabling the skin to have elasticity and reducing wrinkles.

2.3 polypeptide mechanism of action with an effect of promoting elastin production

Palmitoyl hexapeptide-12 (Palmitoyl Hexapeptide-12, CAS number 171263-26-6), derived from Spring fragments of elastin, can promote fibroblast proliferation and elastin synthesis, and improve skin elasticity. In addition, it can induce fibroblasts and monocytes to reach specific locations, promoting wound repair and tissue renewal.

2.4 Polypeptides having a mechanism of action that compensates for proteoglycan deficiency

Acetyl tetrapeptide-9 (Acetyl Tetrapeptide-9) can promote the synthesis of photoprotein glycans. The photoprotein glycan is a member of the family of small-molecule proteoglycans, is an important component of extracellular matrix (ECM), and can regulate collagen fibers to be connected with high efficiency, so that functionalized collagen fibers are formed, and the skin thickness and compactness are maintained. Acetyl tetrapeptide-9 also promotes the synthesis of type I collagen, deeply repairing the skin barrier by simultaneously stimulating the synthesis of laser proteoglycan and type I collagen.

Acetyl tetrapeptide-11 (Acetyl Tetrapeptide-11) stimulates keratinocyte growth and keratinocyte synthesis of syndecan-1.syndecan-1 is a proteoglycan that can strengthen the cohesive force between epidermal cells. Acetyl tetrapeptide-11 also promotes XVII-type collagen production. The type XVII collagen is the main protein of the hemidesmosome and plays an important role in the attachment of the epidermis and the epidermis-dermis junction. Acetyl tetrapeptide-11 remodels and reinforces the skin structure by simultaneously stimulating synthesis of syndecan-1 and XVII type collagen.

Tripeptide-10citrulline (CAS number: 960531-53-7) can compensate for the deficiency of core proteoglycan function caused by aging, can be combined with collagen fibrils to regulate collagen fibril formation, enhance collagen fibril stability, ensure uniform fibril diameter and spatial structure, maintain skin integrity, and make skin soft and elastic.

The invention combines the structural component characteristics of skin, and utilizes different action targets of polypeptide on skin to obtain the water-in-oil skin repair polypeptide composition by dispersing the polypeptide in an oil phase system. The beneficial effects obtained by the invention relative to the prior art include:

(1) The polypeptide is dissolved in a small amount of water phase and dispersed in oil phase to form a water-in-oil system, so that the stability and the use safety of the polypeptide are improved.

(2) The polypeptide is concentrated in the water phase, so that the concentration of the local active ingredients is increased, and the better skin repair effect is achieved.

(3) The oil phase in the system can promote the transdermal absorption of the water-soluble polypeptide, and the same dosage can achieve better skin repair effect.

(4) The invention can achieve better skin repair effect with lower cost, and the preparation process is simple, thus being suitable for industrial mass production.

Drawings

Microscopic image of the water-in-oil system of the polypeptide composition of fig. 1 (40 x 10 times)

FIG. 2 polypeptide cumulative transdermal and cumulative skin hold-up in vitro (24 h)

Detailed Description

For a better understanding of the present invention, the following detailed description of the invention is provided in connection with the accompanying examples and figures, but is not limited to the following examples.

Examples preparation of the compositions of examples 1-5 and comparative examples 1-5

The formulation ratios (percentages) of the compositions of examples 1-5 and comparative examples 1-5 are as follows:

methods for preparing the compositions of examples 1-5 and comparative examples 1-5:

1. accurately weighing the raw materials 5, 6, 7, 8 and 9, adding the raw material 1 with the prescription amount, and fully dissolving;

2. accurately weighing the raw materials 2, 3 and 4, and uniformly mixing the raw materials with the solution obtained in the step 1. Microscopic images of example 3 were observed using a biological digital microscope, as shown in fig. 1, and a number of droplets of uniform size were observed under a microscope at 40 x 10 x and uniformly distributed in the oil phase to form a water-in-oil system.

Test example 1 stability test

1.1 instruments

Constant temperature and humidity box, high Performance Liquid Chromatograph (HPLC)

1.2 test samples

Examples 1 to 5, comparative examples 1 to 5

1.3 test basis

Guidelines for testing stability of crude drug and preparation of four general rules 9001 of 2015 edition of Chinese pharmacopoeia

1.4 test conditions and test items

Acceleration test: the content of the polypeptide in each sample was measured by HPLC at 40.+ -. 2 ℃ and 75%.+ -. 5% RH in a constant temperature and constant humidity cabinet at 1, 2, 3 and 6 months, respectively, to evaluate the stability.

Long-term test: the content of the polypeptide in each sample was measured by HPLC at 25.+ -. 2 ℃ and 60%.+ -. 10% RH in a constant temperature and humidity oven at 3, 6, 9, 12, 18, 24, 36 months, respectively, to evaluate the stability.

1.5 stability test results

The stability data for the samples of examples 1-5, comparative examples 1-5 after 6 months of standing under accelerated test conditions are shown in Table 1 below:

TABLE 1 stability test data (content should be 95% -105% of the indicated amount) accelerated for 6 months

The stability data for the samples of examples 1-5, comparative examples 1-5 after 6 months of standing under long term test conditions are shown in Table 2 below:

TABLE 2 stability test data (content should be 95% -105% of the indicated amount) for long term 6 months

From the results shown in tables 1 and 2, the polypeptide content of example 1 was not significantly changed and was still within the quality standard after 6 months of the acceleration test and the long-term test, while the polypeptide content of comparative example 1 was significantly reduced, indicating that the polypeptide had better stability in water-in-oil systems. Similarly, comparing the results of the different tests of example 2 and comparative example 2, example 3 and comparative example 3, example 4 and comparative example 4, and example 5 and comparative example 5, it is known that the stability of the polypeptides in different types in the water-in-oil system is better than that in the water-based system, the polypeptide content in the water-based system is reduced to different degrees after the acceleration test and the long-term test for 6 months, the reduction of the polypeptide content inevitably leads to the reduction of the efficacy, and even harmful degradation products can be generated, so that the polypeptide has potential harm to human bodies. Therefore, the stability and safety of the active polypeptide can be improved by dispersing the active polypeptide in a water-in-oil system, and a more excellent skin repair effect can be obtained under the condition of the same dosage.

Test example 2 in vitro cumulative transdermal and cumulative skin hold-up test

2.1 instruments

Intelligent drug transdermal diffusion tester and High Performance Liquid Chromatograph (HPLC)

2.2 test samples

Example 1, comparative example 1

2.3 test methods

Samples were evaluated for transdermal properties using vertical Franz diffusion Chi Fa. The isolated skin of the abdomen of SD rat is fixed between the receiving chamber and the supply chamber of the diffusion cell, 1g sample is taken on the skin surface of the supply chamber, and the effective diffusion area is 3.14cm ² Physiological saline is added into the receiving pool as receiving liquid, and the clean bubbles are discharged to ensure that one side of the dermis is completely contacted with the receiving liquid, and the mixture is stirred and diffused at 32 ℃ and 300 r/min. And respectively taking 0.5mL of receiving solution at 4h, 8h, 12h, 16h, 20h and 24h, and timely supplementing the same amount of constant temperature blank receiving solution. The concentration of the polypeptide in the receiving liquid is measured by HPLC, and the accumulated transdermal quantity of the polypeptide per unit area at different times is calculated according to the following formula:

wherein: q (Q) _n To accumulate transdermal flux; c (C) _n The concentration of the polypeptide in the receiving liquid is obtained during the sampling; v is the volume of normal saline in the receiving tank; c (C) _i The concentration of the polypeptide in the receiving liquid is the 1 st time to the last sampling; v (V) _i For each sample volume; a is the effective diffusion area.

After 24 hours, the skin is taken off, the residual liquid of the sample is washed off by ultrapure water, then sheared, ultrapure water is added for homogenization treatment, ultrasound is carried out for 5min, centrifugation is carried out for 10min at 10000r/min, the supernatant is taken and detected by an HPLC method, and the skin retention of the polypeptide unit area is calculated according to the following formula:

Q _s ＝C _s ×V/A

wherein Q is _s Is the accumulated hold-up; c (C) _s The mass concentration of the polypeptide in the skin sample liquid is measured at a sampling time point; v is the volume of the supernatant; a is the effective diffusion area.

2.4 test results

Example 1, comparative example 1 the in vitro cumulative transdermal and cumulative skin hold up of the polypeptides in the samples after 24h transdermal diffusion test are shown in figure 2.

FIG. 2 shows that the cumulative transdermal amount of the polypeptide component of example 1 over 24 hours is 63.19. Mu.g/cm ² The cumulative skin hold-up was 30.28. Mu.g/cm ² The cumulative transdermal amount of the polypeptide component of comparative example 1 over 24 hours was 24.64. Mu.g/cm ² Cumulative skin hold up was 3.97 μg/cm ² . Therefore, the polypeptide is directly dissolved in the water-based system, the water-soluble polypeptide is difficult to penetrate the skin barrier due to the limitation of the skin barrier, the transdermal quantity and the skin retention are low, the polypeptide is easier to be absorbed in the skin by virtue of the similar principle of the oil phase and the skin barrier after being dispersed in the oil phase, and the polypeptide component is dissolved in a small amount of water phase, so that the local polypeptide concentration in the water-in-oil system is higher, and the increase of the concentration gradient is also beneficial to promoting the transdermal absorption. The comparison test result shows that the polypeptide is dispersed in the water-in-oil system, the transdermal quantity and the skin retention are obviously improved, and the skin retention is obviously improvedTo accumulate in the skin to form an active ingredient reservoir, thereby more effectively and permanently exerting a skin repair effect. Test example 3 experiment for promoting collagen formation

3.1 animal test

Healthy 10 weeks nude mice, males, and body weight about 20 g.

3.2 reagents and instruments

Skin hydroxyproline assay kit (institute of bioengineering, nanjing), examples 1-5, comparative examples 1-5, spectrophotometry.

3.3 test methods

The samples of examples 1-5 and comparative examples 1-5 were applied to the dorsum of the right ear of the nude mice, respectively, and 1g of the composition was administered once a day in the morning and evening. The left side was not used as a blank. The same feed was fed under clean, ventilated, dry, room temperature 22±2 ℃.5 medicines are taken after 30 days of administration, and the medicine is taken for acting the part and the skin at the corresponding part on the opposite side.

The wet tissue weight of 45mg is weighed by a balance and put into a clean test tube, 1mL of hydrolysate in a hydroxyproline assay kit is added, and the mixture is uniformly mixed. Covering with boiling water bath for hydrolysis for 20min, and mixing every 10min for more complete hydrolysis. The test tube was removed, cooled, 1 drop of indicator of the kit was added to each tube, and shaken well. Then adding 1mL of the first solution into the kit, adjusting the pH value, and shaking uniformly. Then, the pH-adjusted solution B is sucked by a sample feeder and is added into each test tube drop by drop until the indicator in each tube is seen to turn yellow-green. Distilled water was added to 10mL and mixed well. Taking 3mL of hydrolysate, adding 20-30mg of active carbon, uniformly mixing, centrifuging at 3500 rpm for 10min, and taking 1mL of supernatant.

Clean tubes were taken and 1mL of standard application solution at 5. Mu.g/mL was added as standard tubes. Adding the reagent 1 into each tube, uniformly mixing, standing for 10min, adding the reagent 2, uniformly mixing, standing for 5min, adding the reagent 3, uniformly mixing in a water bath at 60 ℃ for 15min, cooling, centrifuging for 10min, taking the supernatant, and measuring the absorbance value at 550nm and 1cm optical path.

3.4 results determination and analysis

Hydroxyproline is a unique amino acid of collagen, and the content of the collagen is often converted by the content of the hydroxyproline. Spectrophotometry is adopted to measure the content of Hydroxyproline (HYP), and the content of collagen in the skin of the mice is converted according to 14% of HYP in the collagen.

TABLE 3 influence of test samples of groups on collagen production in mouse skin

Note that: * Indicating that each of examples 1-5 was very different from the blank, p <0.001.# indicates that example 1 has statistical significance compared to comparative example 1, p <0.05.# indicates that example 2 is significantly different from comparative example 2, p <0.01.# # # indicates that examples 3-5 differ significantly from the corresponding comparative examples 3-5, respectively, with p <0.001.

As can be seen from the results in table 3, the collagen content in the skin of the mice was greatly increased after the treatment of examples 1, 2, 3, 4 and 5, compared to the control group, and the polypeptide compositions of the examples showed a very significant difference (p < 0.001) compared to the control group, indicating that the polypeptide compositions of the examples were able to significantly promote collagen synthesis. By comparing the examples with corresponding comparative examples, under the condition that the types and the contents of the polypeptides are consistent, the polypeptides can better promote collagen synthesis in a water-in-oil system, and the content of the collagen in the skin of the mice is higher. In addition, under the condition that the concentration of the active ingredients is kept consistent, the content of the collagen increases along with the increase of the types of the polypeptides, which indicates that different polypeptides can promote the synthesis of the collagen from different action targets, and the different polypeptides have synergistic effect, so that the content of the collagen in the skin is obviously increased, and a better skin repair effect is achieved.

Test example 4 aged skin repair test

4.1 subject

330 healthy women aged 50-65 were randomized into 11 groups, with an average of 30 individuals per group.

4.2 test samples

Examples 1-5, comparative examples 1-5, with placebo as a blank.

4.3 test instruments

Cutometer skin elasticity tester (Germany CK company)

4.4 test procedure

The blank value of 30min after the face area is cleaned is measured firstly, then the test sample is uniformly coated on the face, a placebo is coated on one side, the test sample is coated on the other side, the test sample is used twice a day, the test sample is used continuously in the morning and evening, other cosmetics cannot be used, the test sample is continuously used for 4 weeks, and the mechanical properties of the skin of the test area before and after the administration are respectively tested and recorded. Testing of the same subject was performed by the same measurer.

4.5 test results

The skin mechanical properties were evaluated by measuring the skin firmness, skin tension, skin viscoelasticity and skin plasticity index of each sample before and after use using a Cutometer, and the repairing effect of the test sample on aged skin was judged.

The percent improvement in skin firmness, skin tension, skin viscoelasticity, and skin plasticity after 4 weeks of use of the test samples were calculated and the results are given in table 4 below.

Table 4 percent improvement in skin firmness, skin tone, skin viscoelasticity, and skin plasticity (%)

Group of	Skin firmness	Skin tension	Skin adhesiveElasticity of	Skin plasticity
					Placebo	0.92％	-0.59％	0.84％	1.21％
Example 1	18.53％	19.26％	16.82％	25.47％
					Comparative example 1	10.81％	9.95％	11.46％	16.28％
Example 2	22.74％	24.38％	20.94％	30.35％
					Comparative example 2	13.43％	15.62％	14.77％	21.68％
Example 3	31.45％	30.96％	28.57％	39.63％
					Comparative example 3	19.26％	20.58％	19.06％	27.72％
Example 4	34.92％	35.50％	32.83％	44.18％
					Comparative example 4	23.68％	25.93％	22.75％	32.26％
Example 5	39.26％	41.37％	39.45％	50.62％
					Comparative example 5	27.51％	30.25％	28.12％	39.75％

From the data in the table, examples 1-5 and comparative examples 1-5 all showed improved skin firmness, skin tone, skin viscoelasticity and skin plasticity measured after 4 weeks of use relative to placebo. Comparing the test results of example 1 with the test results of comparative example 1, under the condition that the types and the amounts of the polypeptides are the same, the improvement percentages of the skin compactness, the skin tension, the skin viscoelasticity and the skin plasticity indexes of example 1 are more obvious after the application for 4 weeks, and the repairing effect of example 1 on aged skin is better than that of comparative example 1, which indicates that the polypeptides are dispersed in an oil phase system, and can exert better skin repairing effect. Similarly, comparison of the results of the different tests of example 2 and comparative example 2, example 3 and comparative example 3, example 4 and comparative example 4, and example 5 and comparative example 5 shows that the skin repair effect of the polypeptides of different kinds is better in the water-in-oil system than in the water-based system. In addition, under the condition that the concentration of the active ingredients is kept consistent, the improvement range of the mechanical properties of the skin is larger along with the increase of the types of the polypeptides, which indicates that different polypeptides can exert effects from different action targets, and the different polypeptides have obvious synergistic effects, so that better skin repair effect is achieved.

The foregoing is a further detailed description of the invention in connection with the preferred embodiments, but is not intended to limit the practice of the invention to such description. It will be apparent to those skilled in the art that several simple deductions or substitutions can be made without departing from the spirit of the invention, and the invention is not limited to the above-mentioned embodiments.

Claims

1. A water-in-oil skin repair polypeptide composition comprising, in mass percent, 0.085% palmitoyl tripeptide-1, 0.085% palmitoyl tetrapeptide-7, 4.83% purified water, 5% peg-6 caprylic/capric glycerides, 10% polyglycerol-4 laurate, and 80% hydrogenated polyisobutene; the palmitoyl tripeptide-1 and the palmitoyl tetrapeptide-7 are present in the purified water, which forms a water-in-oil system with the PEG-6 caprylic/capric glycerides, the polyglycerol-4 laurate, and the hydrogenated polyisobutene.

2. A water-in-oil skin repair polypeptide composition comprising, in mass percent, 0.05% palmitoyl tripeptide-1, 0.05% palmitoyl tetrapeptide-7, 0.07% hexapeptide-9, 4.83% purified water, 5% peg-6 caprylic/capric glycerides, 10% polyglyceryl-4 laurate, and 80% hydrogenated polyisobutene; the palmitoyl tripeptide-1, the palmitoyl tetrapeptide-7, and the hexapeptide-9 are present in the purified water, which forms a water-in-oil system with the PEG-6 caprylic/capric glycerides, the polyglycerol-4 laurate, and the hydrogenated polyisobutene.

3. A water-in-oil skin repair polypeptide composition comprising, in mass percent concentration, 0.04% palmitoyl tripeptide-1, 0.04% palmitoyl tetrapeptide-7, 0.04% hexapeptide-9, 0.05% palmitoyl hexapeptide-12, 4.83% purified water, 5% peg-6 caprylic/capric glycerides, 10% polyglyceryl-4 laurate, and 80% hydrogenated polyisobutene; the palmitoyl tripeptide-1, the palmitoyl tetrapeptide-7, the hexapeptide-9, and the palmitoyl hexapeptide-12 are present in the purified water, which forms a water-in-oil system with the PEG-6 caprylic/capric glycerides, the polyglycerol-4 laurate, and the hydrogenated polyisobutene.

4. A water-in-oil skin repair polypeptide composition comprising, in mass percent concentration, 0.03% palmitoyl tripeptide-1, 0.03% palmitoyl tetrapeptide-7, 0.03% hexapeptide-9, 0.04% palmitoyl hexapeptide-12, 0.04% acetyl tetrapeptide-9, 4.83% purified water, 5% peg-6 caprylic/capric glycerides, 10% polyglycerol-4 laurate, and 80% hydrogenated polyisobutene; the palmitoyl tripeptide-1, the palmitoyl tetrapeptide-7, the hexapeptide-9, the palmitoyl hexapeptide-12, and the acetyl tetrapeptide-9 are present in the purified water, which forms a water-in-oil system with the PEG-6 caprylic/capric glycerides, the polyglycerol-4 laurate, and the hydrogenated polyisobutene.

5. Use of a water-in-oil skin repair polypeptide composition according to any one of claims 1-4 for the preparation of a skin care product or a medical product for external application to the skin.