Amphiphilic polypeptide assembly as well as preparation method and application thereof

Abstract

The invention relates to an amphiphilic polypeptide assembly as well as a preparation method and application thereof. The amphiphilic polypeptide assembly is obtained by assembling palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 in water. The amphiphilic polypeptide assembly provided by the invention synergistically improves the slow release ability of the three polypeptides, contributes to long-acting exertion of the tightening, anti-wrinkle and anti-inflammatory effects of the three polypeptides on the skin, and has a wide application prospect in the field of cosmetic preparation. The invention further provides a co-assembly containing the amphiphilic polypeptide, a surfactant is additionally added on the basis of the three polypeptides, and the ability of the original polypeptide penetrating through the skin cuticle is further improved.

Description

Technical Field

[0001] The present invention relates to the field of cosmetics, in particular to an amphiphilic polypeptide assembly and a preparation method and application thereof, and specifically to the self-assembly and co-assembly of amphiphilic polypeptide molecules and the application thereof in cosmetics. Background Art

[0002] Polypeptides are compounds formed by amino acids connected by peptide bonds. Amphiphilic polypeptide molecules have amphiphilic properties similar to natural phospholipid molecules. They can spontaneously form micelles in water and self-assemble to form aggregates, in which the lipid tails are wrapped in the core center and the hydrophilic head groups of the polypeptides are exposed on the outer surface. This unique self-assembly structure allows these polypeptide self-assembly materials to play a role in in vitro biological activity and is also related to potential applications in the field of cosmetics. For example, CN103012558A discloses a cationic symmetrical amphiphilic self-assembling polypeptide, which has a good antibacterial effect on Staphylococcus aureus, can be used to develop new nano antibacterial drugs, and can be used as a molecular sensor to detect changes in the microenvironment.

[0003] In skin care products or similar personal care formulations, the presence of peptide self-assembly may provide a reservoir for cosmetic active peptides, which are slowly released in a dynamic equilibrium between monomers and self-assembly structures, thereby maintaining the maximum steady-state concentration of free monomeric peptides absorbed by the stratum corneum, while there is no way to achieve this slow release phenomenon in amphiphilic peptide monomers. In addition, the self-assembled gel formed by amphiphilic peptides can provide wrinkle filling, skin tightening, barrier repair and other skin-friendly effects in skin care products.

[0004] In summary, how to continuously release functional peptides and utilize the assembly structure formed by amphiphilic peptides to allow skin care products to penetrate into the deep skin as much as possible and maximize the efficacy of skin care products has become one of the urgent problems to be solved in this field. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides an amphiphilic polypeptide assembly and a preparation method and application thereof. The raw materials for preparing the amphiphilic polypeptide assembly include palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12. The assembly formed by the above three polypeptides synergistically improves the sustained-release ability of the polypeptides, which helps to long-term exert the firming, anti-wrinkle and anti-inflammatory effects of the three polypeptides on the skin.

[0006] To achieve this object, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides an amphiphilic polypeptide assembly, wherein the amphiphilic polypeptide assembly is assembled in water from palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12.

[0008] Palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 used in the present invention are all amphipathic polypeptides. The palmitoyl group is hydrophobic, and the peptide chain has hydrophilic groups such as amino, carboxyl, and hydroxyl groups, thereby forming an amphipathic polypeptide. Under ultrasonic conditions, the amphipathic polypeptide forms a self-assembly and co-assembly structure. Among them, palmitoyl tripeptide-1 has the effects of promoting collagen synthesis, firming and anti-wrinkle; palmitoyl tetrapeptide-7 has the effect of inhibiting the expression of inflammatory factor IL-6; palmitoyl hexapeptide-12 is an elastin fragment peptide, which has the effect of promoting fibroblasts to synthesize elastin and collagen. The present invention also synergistically improves the sustained-release capacity of the three polypeptides by compounding and using the above three polypeptides, which helps to exert the firming, anti-wrinkle and anti-inflammatory effects of the three polypeptides on the skin for a long time.

[0009] Preferably, the mass percentage of palmitoyl tripeptide-1 in the water is 0.001% to 0.1%, the mass percentage of palmitoyl tetrapeptide-7 is 0.001% to 0.1%, and the mass percentage of palmitoyl hexapeptide-12 is 0.001% to 0.1%.

[0010] The specific point values ​​of the above 0.001% to 0.1% may be 0.001%, 0.005%, 0.01%, 0.02%, 0.04%, 0.05%, 0.06%, 0.08% or 0.1%, etc.

[0011] In a second aspect, the present invention provides a method for preparing the amphiphilic polypeptide assembly as described in the first aspect, the preparation method comprising: dissolving palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 in water to obtain a first mixed solution, and obtaining the amphiphilic polypeptide assembly after ultrasonication.

[0012] Preferably, the first mixed solution contains, in terms of mass percentage, 0.001% to 0.1% palmitoyl tripeptide-1, 0.001% to 0.1% palmitoyl tetrapeptide-7 and 0.001% to 0.1% palmitoyl hexapeptide-12.

[0013] Preferably, the pH of the first mixed solution is 5-8 (eg, 5, 6, 6.5, 7 or 8, etc.).

[0014] Preferably, the power of the ultrasound is 60-120 W (for example, 60 W, 70 W, 80 W, 90 W, 100 W, 110 W or 120 W).

[0015] Preferably, the ultrasound time is 15 to 25 min (for example, 15 min, 17 min, 19 min, 20 min, 21 min, 23 min or 25 min, etc.).

[0016] Preferably, the temperature of the ultrasound is 25-85°C (for example, it can be 25°C, 30°C, 40°C, 50°C, 55°C, 60°C, 70°C, 80°C or 85°C, etc.), preferably 40-60°C (for example, it can be 40°C, 45°C, 48°C, 50°C, 52°C, 55°C or 60°C, etc.).

[0017] In a third aspect, the present invention provides a co-assembly containing an amphiphilic polypeptide, wherein the raw materials for preparing the co-assembly include palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, palmitoyl hexapeptide-12 and a surfactant, wherein the surfactant includes any one of hydrogenated lecithin, Tween-20, Tween-60 or span-85, or a combination of at least two of them.

[0018] Preferably, the surfactant is hydrogenated lecithin and Tween-20.

[0019] The present invention adds two surfactants to the three polypeptides of palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 to form a co-assembly structure containing amphiphilic polypeptides, thereby improving the sustained-release ability of the three polypeptides. Hydrogenated lecithin and Tween-20 have the ability to synergistically enhance the ability of polypeptides to penetrate the stratum corneum of the skin, and enhance the retention amount and stability of polypeptides in the skin, thereby improving the ability of polypeptide activity.

[0020] Preferably, the mass ratio of the hydrogenated lecithin to Tween-20 is (3-7):1.

[0021] The specific point values ​​of the above (3-7) can be selected from 3, 4, 4.5, 5, 5.5, 6 or 7, etc.

[0022] Preferably, the co-assembly is prepared by a method comprising the following steps: dissolving the surfactant in water, mixing it with the palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 to obtain a second mixed solution, and stirring to obtain the co-assembly.

[0023] Preferably, the co-assembly is prepared by a method comprising the following steps: completely dissolving the surfactant in water at 75 to 85°C (for example, 75°C, 77°C, 79°C, 80°C, 81°C, 83°C or 85°C, etc.), reducing the temperature to 55 to 65°C (for example, 55°C, 57°C, 59°C, 60°C, 61°C, 63°C or 65°C, etc.), mixing with the palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 to obtain a second mixed solution, and stirring to obtain the co-assembly.

[0024] Preferably, the second mixed solution contains, by mass percentage, 0.001% to 0.1% (palmitoyl tripeptide-1, 0.001% to 0.1% palmitoyl tetrapeptide-7, 0.001% to 0.1% palmitoyl hexapeptide-12 and 0.001% to 3% surfactant.

[0025] The specific point values ​​of the above 0.001% to 0.1% may be 0.001%, 0.005%, 0.01%, 0.02%, 0.04%, 0.05%, 0.06%, 0.08% or 0.1%, etc.

[0026] The specific point values ​​of 0.001% to 3% mentioned above can be selected from 0.001%, 0.005%, 0.01%, 0.02%, 0.04%, 0.05%, 0.06%, 0.08%, 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5% or 3%, etc.

[0027] In a fourth aspect, the present invention provides a composite nanoparticle encapsulating an active substance, wherein the composite nanoparticle contains the amphiphilic polypeptide assembly described in the first aspect or the co-assembly containing the amphiphilic polypeptide described in the third aspect.

[0028] The amphiphilic polypeptide assembly and the co-assembly containing the amphiphilic polypeptide provided by the present invention can be used to encapsulate active substances, promote the penetration of active substances into deep layers of the skin, and thus maximize the efficacy of the active substances.

[0029] Preferably, the composite nanoparticles further contain active substances.

[0030] Preferably, the active substance comprises any one of hydroxytyrosol, β-glucan or soluble collagen, or a combination of at least two thereof.

[0031] Preferably, the composite nanoparticles are prepared by a method comprising the following steps: uniformly dispersing the active substance in an aqueous solution, mixing with the amphiphilic polypeptide assembly described in the first aspect or the co-assembly containing the amphiphilic polypeptide described in the third aspect, to obtain the composite nanoparticles.

[0032] Preferably, the added amount of the active substance is 0.01-2% (for example, it can be 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 0.7%, 1%, 1.5% or 2%, etc.).

[0033] In a fifth aspect, the present invention provides the use of any one or a combination of at least two of the amphiphilic polypeptide assembly as described in the first aspect, the preparation method of the amphiphilic polypeptide assembly as described in the second aspect, the co-assembly containing amphiphilic polypeptides as described in the third aspect, or the composite nanoparticles encapsulating active substances as described in the fourth aspect in the preparation of cosmetics.

[0034] Preferably, the cosmetics include any one of lotion, toner, eye cream, essence milk, face cream or face mask, or a combination of at least two of them.

[0035] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] (1) The present invention provides an amphiphilic polypeptide assembly containing palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12, which synergistically enhances the sustained-release ability of the three polypeptides and helps to exert the firming, anti-wrinkle and anti-inflammatory effects of the three polypeptides on the skin for a long time.

[0038] (2) The present invention also provides a co-assembly containing an amphiphilic polypeptide, in which a surfactant is additionally added to the above three polypeptides, thereby further enhancing the ability of the original polypeptide to penetrate the stratum corneum of the skin.

[0039] (3) The present invention uses an amphiphilic polypeptide assembly or a co-assembly containing an amphiphilic polypeptide to encapsulate the active substance, thereby promoting the penetration of the active substance into the deep layer of the skin, thereby maximizing the efficacy of the active substance, and has broad application prospects in the field of cosmetic preparation. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 This is the TEM test structure diagram of Example 1.

[0041] Figure 2 This is the TEM test structure diagram of Example 8. DETAILED DESCRIPTION

[0042] To further illustrate the technical means and effects of the present invention, the present invention is further described below in conjunction with the embodiments and drawings. It should be understood that the specific implementation methods described herein are only used to explain the present invention, rather than to limit the present invention.

[0043] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0044] Example 1

[0045] In this embodiment, an amphiphilic polypeptide assembly is prepared, and the preparation method comprises the following steps: 0.05 g of palmitoyl tripeptide-1, 0.05 g of palmitoyl tetrapeptide-7 and 0.05 g of palmitoyl hexapeptide-12 are dissolved in 100 mL of deionized water to obtain a first mixed solution with a pH of 6.5, and ultrasonicated in a water bath at 50° C. for 20 min with an ultrasonic power of 90 W to obtain the amphiphilic polypeptide assembly.

[0046] The amphiphilic polypeptide assembly was subjected to TEM testing, the testing method comprising: dripping the solution onto a carbon film copper grid at 200 KV, soaking it in phosphotungstic acid (2%) and then drying it for testing. Figure 1 As shown, it can be seen that the three polypeptides aggregate to form a unique network-like assembly structure.

[0047] Example 2

[0048] In this embodiment, an amphiphilic polypeptide assembly is prepared, and the preparation method comprises the following steps: 0.09 g of palmitoyl tripeptide-1, 0.01 g of palmitoyl tetrapeptide-7 and 0.09 g of palmitoyl hexapeptide-12 are dissolved in 100 mL of deionized water to obtain a first mixed solution with a pH of 8, and ultrasonicated in a water bath at 40° C. for 25 min with an ultrasonic power of 60 W to obtain the amphiphilic polypeptide assembly.

[0049] Example 3

[0050] In this embodiment, an amphiphilic polypeptide assembly is prepared, and the preparation method comprises the following steps: 0.01 g of palmitoyl tripeptide-1, 0.09 g of palmitoyl tetrapeptide-7 and 0.01 g of palmitoyl hexapeptide-12 are dissolved in 100 mL of deionized water to obtain a first mixed solution with a pH of 5, and ultrasonicated in a water bath at 60° C. for 15 min with an ultrasonic power of 120 W to obtain the amphiphilic polypeptide assembly.

[0051] Example 4

[0052] This example prepares an amphiphilic polypeptide assembly, which differs from Example 1 only in that the temperature of the water bath is 38°C.

[0053] Example 5

[0054] This example prepares an amphiphilic polypeptide assembly, which differs from Example 1 only in that the temperature of the water bath is 62°C.

[0055] Example 6

[0056] This example prepares an amphiphilic polypeptide assembly, which differs from Example 1 only in that the ultrasonication time is 12 minutes.

[0057] Example 7

[0058] This example prepares an amphiphilic polypeptide assembly, which differs from Example 1 only in that the ultrasonication time is 28 min.

[0059] Example 8

[0060] This embodiment prepares a co-assembly containing an amphiphilic polypeptide, and the preparation method includes the following steps: dissolving hydrogenated lecithin and Tween-20 in 500 mL of deionized water, dissolving completely under 80°C water bath conditions, reducing the temperature to 60°C, adding palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12. In terms of mass percentage, the mixed solution contains 0.008% palmitoyl tripeptide-1, 0.008% palmitoyl tetrapeptide-7, 0.008% palmitoyl hexapeptide-12, 0.06% hydrogenated lecithin and 0.012% Tween-20. The stirred mixed solution is a slightly blue transparent solution, and the co-assembly is obtained.

[0061] The co-assembly was subjected to TEM testing, the testing method comprising: dripping the solution onto the carbon film copper grid at 200KV, soaking it in phosphotungstic acid (2%) and then drying it for testing. Figure 2 As shown, it can be seen that the three polypeptides, hydrogenated phosphatidylcholine and Tween-20 co-assemble to form a unique ball-and-stick assembly structure.

[0062] Example 9

[0063] This embodiment prepares a co-assembly containing an amphiphilic polypeptide, and the preparation method includes the following steps: dissolving hydrogenated lecithin and Tween-20 in 500 mL of deionized water, dissolving completely under 85°C water bath conditions, reducing the temperature to 55°C, and adding palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, and palmitoyl hexapeptide-12. In terms of mass percentage, the mixed solution contains 0.02% palmitoyl tripeptide-1, 0.002% palmitoyl tetrapeptide-7, 0.02% palmitoyl hexapeptide-12, 0.75% hydrogenated lecithin, and 0.25% Tween-20. The stirred mixed solution is a slightly blue transparent solution, and the co-assembly is obtained.

[0064] Example 10

[0065] This embodiment prepares a co-assembly containing an amphiphilic polypeptide, and the preparation method includes the following steps: dissolving hydrogenated lecithin and Tween-20 in 500 mL of deionized water, dissolving completely under 75°C water bath conditions, reducing the temperature to 65°C, adding palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12. In terms of mass percentage, the mixed solution contains 0.002% palmitoyl tripeptide-1, 0.02% palmitoyl tetrapeptide-7, 0.002% palmitoyl hexapeptide-12, 0.035% hydrogenated lecithin and 0.005% Tween-20. The stirred mixed solution is a slightly blue transparent solution, and the co-assembly is obtained.

[0066] Comparative Example 1

[0067] In this comparative example, an amphiphilic polypeptide assembly was prepared, which differed from Example 1 only in that palmitoyl tripeptide-1 was not added, and the proportion of palmitoyl tripeptide-1 was distributed to palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 in proportion.

[0068] Comparative Example 2

[0069] In this comparative example, an amphiphilic polypeptide assembly was prepared, which differed from Example 1 only in that palmitoyl tetrapeptide-7 was not added, and the proportion of palmitoyl tetrapeptide-7 was distributed to palmitoyl tripeptide-1 and palmitoyl hexapeptide-12 in proportion.

[0070] Comparative Example 3

[0071] In this comparative example, an amphiphilic polypeptide assembly was prepared, which differed from Example 1 only in that palmitoyl hexapeptide-12 was not added, and the proportion of palmitoyl hexapeptide-12 was proportionally distributed to palmitoyl tripeptide-1 and palmitoyl tetrapeptide-7.

[0072] Comparative Example 4

[0073] This comparative example prepared an amphiphilic polypeptide assembly, which differed from Example 1 only in that palmitoyl tripeptide-1 was replaced with palmitoyl tripeptide-5 of equal mass.

[0074] Comparative Example 5

[0075] This comparative example prepared an amphiphilic polypeptide assembly, which differed from Example 1 only in that palmitoyl tetrapeptide-7 was replaced by palmitoyl tripeptide-8 of equal mass.

[0076] Comparative Example 6

[0077] This comparative example prepared an amphiphilic polypeptide assembly, which differed from Example 1 only in that palmitoyl hexapeptide-12 was replaced by palmitoyl hexapeptide-14 of equal mass.

[0078] Comparative Example 7

[0079] In this comparative example, a solution containing an amphiphilic polypeptide was prepared, and the preparation method included the following steps: 0.05 g of palmitoyl tripeptide-1, 0.05 g of palmitoyl tetrapeptide-7 and 0.05 g of palmitoyl hexapeptide-12 were dissolved in 100 mL of deionized water, and the pH was adjusted to 6.5.

[0080] Comparative Example 8

[0081] This comparative example prepares a co-assembly containing an amphiphilic polypeptide, which differs from Example 8 only in that Tween-20 is replaced by an equal amount of Tween-60.

[0082] Comparative Example 9

[0083] This comparative example prepares a solution containing an amphiphilic polypeptide, which is different from Example 8 only in that after the three polypeptides are added, they are simply mixed and no co-assembly is formed.

[0084] Application Example 1

[0085] This application example prepares a composite nanoparticle encapsulating an active substance, and the preparation method includes the following steps: 0.5% soluble collagen is uniformly dispersed in an aqueous solution, and then the amphiphilic polypeptide assembly provided in Example 1 is mixed therewith to obtain composite nanoparticles encapsulating soluble collagen.

[0086] Application Example 2-7

[0087] This application example prepares a composite nanoparticle encapsulating an active substance, which is different from application example 1 only in that the amphiphilic polypeptide assembly provided in examples 2-7 is mixed with soluble collagen.

[0088] Application Examples 8-10

[0089] This application example prepares a composite nanoparticle encapsulating an active substance, which is different from application example 1 only in that the co-assembly containing the amphiphilic polypeptide provided in Examples 8-10 is mixed with soluble collagen.

[0090] Comparative Application Examples 1-9

[0091] This application example prepares a composite nanoparticle encapsulating an active substance, and the only difference from application example 1 is that the product prepared in comparative examples 1-9 is mixed with soluble collagen.

[0092] Test Example 1

[0093] This test example evaluates the skin tightening effect of the amphiphilic polypeptide assembly or the co-assembly containing the amphiphilic polypeptide provided in the examples and comparative examples, and includes the following test steps:

[0094] (1) 190 trial subjects aged 20-45 were randomly selected and divided into 19 groups, with 10 subjects in each group. The trial subjects used the products provided in the embodiments and comparative examples (once in the morning and once in the evening) for 30 days. During the trial period, the subjects were not allowed to use other items that maintain skin elasticity and cosmetics that affect the results. During the trial period, the subjects did not change their original eating habits.

[0095] (2) Use the skin elasticity meter Cutometer MPA580 to test the skin condition on the 0th day and the 30th day respectively. The test site is the zygomatic area. During the measurement, the probe repeats the measurement 3 times in the same test area and takes the average value. The test parameter is R2 (the ratio of the skin rebound amount Ua when there is no negative pressure to the maximum stretch amount Uf when there is negative pressure). The closer R2 is to 1, the better the elasticity of the skin. The average change value of R2 = R2 value on the 30th day - R2 value on the 0th day, and the average change rate of R2 = R2 average change value / R2 value on the 0th day. The test results are shown in Table 1.

[0096] Table 1

[0097]

[0098] Test Example 2

[0099] This test example evaluates the anti-wrinkle effect of the amphiphilic polypeptide assembly or the co-assembly containing the amphiphilic polypeptide provided in the examples and comparative examples, and includes the following test steps:

[0100] (1) 190 trial subjects in Test Example 1 were randomly divided into 19 groups, with 10 subjects in each group. The products provided in the embodiment and the comparative example were used respectively (once in the morning and once in the evening) for 90 consecutive days.

[0101] (2) Follow-up visits were conducted on the 30th day (D30) and the 90th day (D90) of the trial, and the same subjects were measured by the same personnel to count the number and length of facial wrinkles, and the improvement of skin wrinkles was statistically analyzed based on the evaluation criteria.

[0102] Evaluation criteria: 10 points, no wrinkles; 7-9 points, 2-3 wrinkles, less than 1.5 cm in length; 4-6 points, 3-6 wrinkles, less than 3 cm in length; 1-3 points, several major wrinkles, more than 4 cm in length, accompanied by shallow wrinkles. The number of people with each score is shown in Table 2.

[0103] Table 2

[0104]

[0105]

[0106] Test Example 3

[0107] This test example tests the anti-inflammatory effect of the amphiphilic polypeptide assembly or the co-assembly containing the amphiphilic polypeptide provided in the examples and comparative examples, and includes the following test steps:

[0108] (1) Raw264.7 cells (mouse macrophages, which have similar functions to primary macrophages and are often used in the study of anti-inflammation, infection, immune response, etc.) in good culture state were diluted to a concentration of 4×10 5 cfu / mL.

[0109] (2) Prepare a 12-well cell plate and add a concentration of 4×10 5 cfu / mL of Raw264.7 cells were transferred to a 12-well plate using a pipette, 1 mL was added to each well, and cultured at 37°C for 24 h, and the culture medium was aspirated.

[0110] (3) Add 0.5 mL of the following samples respectively: blank group (culture medium containing 2% serum), LPS group (addition of 2 μg / mL lipopolysaccharide), test group (addition of sample solutions of the examples and comparative examples containing 2 μg / mL lipopolysaccharide), and positive control group (addition of 2 μg / mL lipopolysaccharide and 0.2 mg / mL dexamethasone), and culture at 37°C for 24 h.

[0111] (4) The supernatant was centrifuged and the concentration of inflammatory factor IL-6 was detected according to the test steps on the ELISA kit. Each group was measured 5 times in parallel and the average value was taken. The results are shown in Table 3.

[0112] Table 3

[0113]

[0114]

[0115] It can be seen from Tables 1 to 3 that:

[0116] (1) By comparing Example 1 with Comparative Example 7, it can be seen that the assembly containing palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 has long-lasting skin tightening, skin elasticity improvement, long-lasting anti-wrinkle and anti-inflammatory effects, while the polypeptide mixture that does not form an assembly has a poor effect.

[0117] (2) Comparing Example 1 with Comparative Examples 1-6, it can be seen that palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 have the ability to synergistically enhance long-term skin firming, improve skin elasticity, and have long-term anti-wrinkle and anti-inflammatory effects.

[0118] (3) Comparing Example 1 with Examples 4-7, it can be seen that the water bath temperature and ultrasonic time during the preparation of the assembly will affect the structure of the assembly, thereby affecting the release of the polypeptide.

[0119] (4) By comparing Example 8 with Comparative Example 9, it can be seen that the co-assembly containing palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, palmitoyl hexapeptide-12, hydrogenated lecithin and Tween-20 has long-lasting skin tightening, skin elasticity enhancement, long-lasting anti-wrinkle and anti-inflammatory effects, while the polypeptide mixture that does not form an assembly has a poor effect.

[0120] (5) By comparing Example 8 with Comparative Example 8, it can be seen that hydrogenated lecithin and Tween-20 have the ability to synergistically stabilize the co-assembly structure, and the stabilization ability is weakened after Tween-20 is replaced by Tween-60.

[0121] Test Example 4

[0122] This test example conducted a transdermal absorption test on the products containing soluble collagen prepared in the application example and the comparative application example, including the following test steps:

[0123] Using piglet skin as a model, an in vitro skin permeation experiment was conducted in a Franz diffusion cell at 36°C and 100 r / min to detect the cumulative transdermal amount (μg / cm 2 ). The results are shown in Table 4.

[0124] Table 4

[0125]

[0126]

[0127] Test Example 5

[0128] In this test example, the products containing soluble collagen prepared in the application example and the comparative application example were subjected to anti-aging tests. Soluble collagen has anti-aging effects, and one manifestation of skin aging is skin sagging. Testing skin elasticity can reflect the penetration effect of soluble collagen in the present invention. This test example evaluated the skin tightening effect with reference to test example 1, and the test results are shown in Table 5.

[0129] Table 5

[0130]

[0131] It can be seen from Table 4 and Table 5 that:

[0132] (1) Comparing Application Example 1 with Comparative Application Example 7, it can be seen that the assembly containing palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 has stronger penetration ability, which can help soluble collagen penetrate into the deep skin and improve skin elasticity.

[0133] (2) Comparing Application Example 1 with Comparative Application Examples 1-6, it can be seen that palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 have the ability to synergistically promote the penetration of polypeptides and active substances included.

[0134] (3) Comparing Application Example 1 with Application Examples 4-7, it can be seen that the water bath temperature and ultrasonic time during the preparation of the assembly will affect the structure of the assembly, thereby affecting the ability of the polypeptide to encapsulate the active substance.

[0135] (4) Comparing Application Example 8 with Comparative Application Example 9, it can be seen that the co-assembly containing palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, palmitoyl hexapeptide-12, hydrogenated lecithin and Tween-20 has a stronger penetration ability, which can help soluble collagen penetrate into the deep skin and improve skin elasticity.

[0136] (5) Comparison of Application Example 8 with Comparative Application Example 8 shows that hydrogenated lecithin and Tween-20 have the ability to synergistically stabilize the structure of the co-assembly. After Tween-20 is replaced with Tween-60, the stabilization ability is weakened and the ability to encapsulate active substances is reduced.

[0137] Test Example 6

[0138] This test example conducts safety evaluation tests on the products provided in the embodiments, comparative examples, application examples and comparative application examples. 40 women aged 18-35 years old with no history of allergies were selected, and each subject tried the sample. After cleaning the back of the subject, apply the spot tester with the sample to the selected position on the back with a non-irritating tape. After pasting, press it lightly with your fingers to make it evenly attached to the skin for 48 hours. The subject keeps the patch area dry within 48 hours, avoids intense exercise, scratching the patch area, and long-term sunlight exposure. After 48 hours, remove the tester and mark it. After 30 minutes, wait for the indentation to disappear and make a judgment under sufficient light.

[0139] The test results showed that all the subjects had negative reactions, no irritation to the skin, and no positive reactions, indicating that the product provided by the present invention is safe and mild.

[0140] In summary, the amphiphilic polypeptide assembly provided by the present invention synergistically improves the sustained-release ability of the three polypeptides, helps to exert the firming, anti-wrinkle and anti-inflammatory effects of the three polypeptides on the skin for a long time, and has broad application prospects in the field of cosmetic preparation.

[0141] The applicant declares that the above is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed by the present invention shall fall within the protection scope and disclosure scope of the present invention.

Claims

1. An amphiphilic polypeptide assembly, characterized in that: The amphiphilic polypeptide assembly is obtained by assembling palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 in water.

2. The amphiphilic polypeptide assembly according to claim 1, characterized in that The mass percentage of palmitoyl tripeptide-1 in the water is 0.001% to 0.1%, the mass percentage of palmitoyl tetrapeptide-7 is 0.001% to 0.1%, and the mass percentage of palmitoyl hexapeptide-12 is 0.001% to 0.1%.

3. A method for preparing the amphiphilic polypeptide assembly according to claim 1 or 2, characterized in that: The preparation method comprises: dissolving palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 in water to obtain a first mixed solution, and obtaining the amphiphilic polypeptide assembly after ultrasonic treatment; Preferably, the pH of the first mixed solution is 5 to 8; Preferably, the power of the ultrasound is 60 to 120 W; Preferably, the ultrasound duration is 15 to 25 minutes; Preferably, the temperature of the ultrasound is 25-85°C, preferably 40-60°C.

4. A co-assembly containing an amphiphilic polypeptide, characterized in that: The raw materials for preparing the co-assembly include palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, palmitoyl hexapeptide-12 and a surfactant, wherein the surfactant includes any one of hydrogenated lecithin, Tween-20, Tween-60 or span-85 or a combination of at least two thereof.

5. The co-assembly according to claim 4, characterized in that: The surfactant is hydrogenated lecithin and Tween-20; Preferably, the mass ratio of the hydrogenated lecithin to Tween-20 is (3-7):

1.

6. The co-assembly according to claim 4 or 5, characterized in that: The co-assembly is prepared by a method comprising the following steps: dissolving the surfactant in water, mixing with the palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 to obtain a second mixed solution, and stirring to obtain the co-assembly; Preferably, the co-assembly is prepared by a method comprising the following steps: completely dissolving the surfactant in water at 75 to 85° C., lowering the temperature to 55 to 65° C., mixing with the palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and palmitoyl hexapeptide-12 to obtain a second mixed solution, and stirring to obtain the co-assembly; Preferably, the second mixed solution contains, in terms of mass percentage, 0.001% to 0.1% palmitoyl tripeptide-1, 0.001% to 0.1% palmitoyl tetrapeptide-7, 0.001% to 0.1% palmitoyl hexapeptide-12 and 0.001% to 3% surfactant.

7. A composite nanoparticle encapsulating an active substance, characterized in that: The composite nanoparticles contain the amphiphilic polypeptide assembly according to claim 1 or 2 or the co-assembly containing the amphiphilic polypeptide according to any one of claims 4 to 6.

8. The composite nanoparticle according to claim 7, characterized in that The composite nanoparticles also contain an active substance; Preferably, the active substance comprises any one or a combination of at least two of hydroxytyrosol, β-glucan or soluble collagen; Preferably, the composite nanoparticles are prepared by a method comprising the following steps: uniformly dispersing the active substance in an aqueous solution, mixing with the amphiphilic polypeptide assembly according to claim 1 or 2 or the co-assembly containing the amphiphilic polypeptide according to any one of claims 4 to 6, to obtain the composite nanoparticles. Preferably, the added amount of the active substance is 0.01-2%.

9. Use of any one or a combination of at least two of the amphiphilic polypeptide assembly according to claim 1 or 2, the method for preparing the amphiphilic polypeptide assembly according to claim 3, the co-assembly containing amphiphilic polypeptides according to any one of claims 4 to 6, or the composite nanoparticles encapsulating active substances according to claim 7 or 8 in the preparation of cosmetics.

10. The use according to claim 9, characterized in that: The cosmetics include any one of lotion, toner, eye cream, essence milk, face cream or face mask, or a combination of at least two of them.

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