Cell active peptide composition for promoting regeneration of multi-type collagen structure protein, preparation method of cell active peptide composition and application of cell active peptide composition in medicine and cosmetology

Abstract

The invention relates to the field of medicine and cosmetology, and discloses a cell active peptide composition for promoting regeneration of multi-type collagen structure protein, a preparation method of the cell active peptide composition and application of the cell active peptide composition in medicine and cosmetology. The composition is prepared from the following raw materials: tripeptide-10 citrulline, hexapeptide-9, hydrolyzed sodium hyaluronate, gamma-aminobutyric acid, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, palmitoyl pentapeptide-4 and the like. The composition can promote the synergistic expression of I-type, III-type, IV-type, VII-type and XVII-type collagen, fibronectin, elastin and glycosaminoglycan of the skin; collagen crosslinking is prevented, a triple-helix structure is maintained, and collagen degradation is inhibited. When being applied to skin medicine and beauty treatment, the composition can accelerate regeneration of structural proteins such as skin collagen, repair skin injury, reduce wrinkles such as crow's feet, lacrimal ducts, nasolabial folds and subocular wrinkles, solve the problem of skin aging caused by loss of the structural proteins such as collagen, and improve skin elasticity and compactness.

Description

Technical Field

[0001] The present invention relates to the fields of medicine and cosmetology, and in particular to a cell active peptide composition for promoting the regeneration of multi-type collagen structural protein, a preparation method thereof, and application thereof in medicine and cosmetology. Background Art

[0002] Collagen is the main component of the extracellular matrix of the skin, and plays a vital role in maintaining the plumpness and elasticity of the skin and keeping the skin youthful overall. However, as time goes by, the collagen content in the skin gradually decreases. This natural physiological process can cause signs of skin aging, such as sagging and wrinkles. In addition, reactive oxygen species (ROS) are one of the key factors that accelerate the loss of collagen and promote skin aging. During the process of photoaging, ultraviolet radiation stimulates the generation of ROS in the skin, which in turn accelerates the degradation of collagen, destroys the balance of the extracellular matrix, and reduces the body's collagen reserves.

[0003] In human skin, multiple types of collagen work together, including types I, III, IV, VII, and XVII. Type I collagen is the most abundant type in the skin, accounting for 60% to 80% of the total collagen content in the dermis, providing the skin with primary strength and support. Close behind is type III collagen, which accounts for 15% to 25% of the collagen in the dermis and plays an important role in the skin's flexibility and repair ability. Together, these two types of collagen form the basis of the structural fibers in the dermis, supporting the integrity of the skin's structure.

[0004] There are multiple ways to supplement collagen, including exogenous supplementation, reducing collagen degradation, and endogenous collagen regeneration. Exogenous supplementation mainly takes the form of injection, injecting recombinant humanized collagen or animal-derived collagen; reducing collagen degradation mainly inhibits the activity of metalloproteinases, and preventing protein cross-linking through antioxidant and anti-glycation methods; endogenous collagen regeneration is achieved through the targeted action of cell active peptides, stimulating autologous cells to regenerate and synthesize collagen and other structural proteins, solving the root cause of structural protein loss.

[0005] Although collagen supplementation has potential benefits in combating skin aging, the current collagen supplementation methods on the market still face some challenges. For example, collagen derived from animals may have the risk of immunogenicity and disease transmission. Although recombinant humanized collagen has advantages in reducing immunogenicity, it still has many problems in terms of structural integrity, type richness, purity, stability, and price. There is little data on its application in human skin and there are many uncertainties. In addition, recombinant collagen and animal collagen have large molecular weights and are difficult to penetrate the epidermis to reach the dermis, so they cannot be effective when used in topical products.

[0006] Therefore, providing a composition that can promote collagen production is a technical problem that needs to be solved at present. Summary of the invention

[0007] In view of the above technical problems, the inventors proposed the technical solution of the present invention, which includes a cell active peptide composition for promoting the regeneration of multi-type collagen structural protein, a preparation method thereof, and applications in medicine and cosmetology.

[0008] The cell active peptide composition of the present invention comprises: tripeptide-10 citrulline, hexapeptide-9, hydrolyzed sodium hyaluronate, γ-aminobutyric acid, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, palmitoyl pentapeptide-4 and other raw materials.

[0009] Among them, hexapeptide-9 can promote type I collagen and type IV collagen. Palmitoyl dipeptide-7 can inhibit the activity of metalloproteinases, increase the expression of structural proteins and hyaluronic acid in the dermis and basement membrane. Tripeptide-10 citrulline can ensure the uniformity of fiber diameter, making collagen fibers better coagulated, thereby increasing the softness of the skin. Acetyl tetrapeptide-3 acts on fibroblasts to accelerate the synthesis of extracellular matrix proteins such as laminin, collagen III and VII. Acetyl tetrapeptide-11 reshapes and strengthens the skin structure by stimulating the synthesis of syndecan-1 and type XVII collagen at the same time. Palmitoyl pentapeptide-4 is the 212-216 sequence of type I collagen, which can increase the expression of type III collagen by fibroblasts. Decarboxylated carnosine hydrochloride can act on glycosylated protein products, prevent further cross-linking of glycosylated proteins, effectively inhibit advanced glycation end products AGES, and protect collagen. γ-aminobutyric acid: It can promote the synthesis of elastin and the formation of elastic fibers in normal skin fibroblasts, thereby improving skin elasticity and having a good effect. Hydrolyzed sodium hyaluronate covalently binds to core protein to form a complex sugar, which plays an important role in skin structure and function.

[0010] The composition of the present invention uses tripeptide-10 citrulline, hexapeptide-9, hydrolyzed sodium hyaluronate, γ-aminobutyric acid, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, and palmitoyl pentapeptide-4 to promote endogenous production of collagen; at the same time, the present invention uses a compound penetration enhancer to further promote endogenous production of collagen in the skin; the composition prepared from the raw materials of the present invention can well supplement skin collagen from the endogenous source and has an excellent anti-aging effect.

[0011] On the one hand, the cell active peptide composition provided by the present invention contains, by mass percentage, 0.0001-1% tripeptide-10 citrulline, 0.0001-0.5% hexapeptide-9, 0.0001-0.5% hydrolyzed sodium hyaluronate, 0.0001-2% γ-aminobutyric acid, 0.0001-1.5% decarboxylated carnosine hydrochloride, 0.0001-1% acetyl tetrapeptide-3, 0.0001-1% acetyl tetrapeptide-11, 0.0001-0.5% palmitoyl dipeptide-7, 0.0001-0.1% palmitoyl pentapeptide-4, 0.5-5% penetration enhancer and the remainder water.

[0012] Furthermore, the amount of the tripeptide-10 citrulline in the composition is in the range of 0.001-0.9%, 0.005-0.9%, 0.01-0.85%, 0.02-0.8%, 0.05-0.75%, etc.

[0013] Furthermore, the amount of hexapeptide-9 in the composition is in the range of 0.001-0.48%, 0.002-0.45%, 0.01-0.43%, 0.02-0.42%, 0.01-0.4%, 0.05-0.4%, etc.

[0014] Furthermore, the amount of the hydrolyzed sodium hyaluronate in the composition is in the range of 0.001-0.48%, 0.002-0.45%, 0.01-0.43%, 0.02-0.42%, 0.01-0.4%, 0.05-0.4%, etc. The molecular weight of the hydrolyzed sodium hyaluronate is less than 1000Da.

[0015] Furthermore, the amount of γ-aminobutyric acid in the composition is in the range of 0.001-1.2%, 0.001-0.6%, 0.002-0.55%, 0.01-0.55%, 0.05-0.5%, 0.06-0.45%, etc.

[0016] Furthermore, in order to more efficiently promote the production of collagen and achieve anti-aging effects, the present invention creatively selects the mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid to be 2-4:1; further selected as a ratio of 2.2:1, 2.5:1, 3:1, 3.5:1 or a range between any two ratios.

[0017] Furthermore, the amount of decarboxylated carnosine hydrochloride in the composition is in the range of 0.001-1.4%, 0.005-1.3%, 0.01-1.2%, 0.02-1%, 0.02-0.8%, etc.

[0018] Furthermore, the amount of acetyl tetrapeptide-3 in the composition is in the range of 0.001-0.9%, 0.005-0.9%, 0.01-0.85%, 0.02-0.8%, 0.05-0.75%, etc.

[0019] Furthermore, the amount of acetyl tetrapeptide-11 in the composition is in the range of 0.001-0.9%, 0.005-0.9%, 0.01-0.85%, 0.02-0.8%, 0.05-0.75%, etc.

[0020] Furthermore, the amount of palmitoyl dipeptide-7 in the composition is in the range of 0.001-0.48%, 0.002-0.45%, 0.005-0.42%, 0.01-0.4%, 0.03-0.35%, etc.

[0021] Furthermore, the amount of palmitoyl pentapeptide-4 in the composition is in the range of 0.0005-0.1%, 0.001-0.095%, 0.005-0.92%, 0.008-0.09%, 0.01-0.09%, etc.

[0022] Further, the amount of the penetration enhancer in the composition is in the range of 1-4.5%, 1.5-4.3%, 1.8-4.2%, 2-4%, etc. The penetration enhancer is dimethyl isosorbide and / or inositol. Further, the penetration enhancer is selected to be a combination of dimethyl isosorbide and inositol in a mass ratio of 1-2:1. Under this condition, the penetration enhancer can efficiently deliver the active ingredient and be efficiently absorbed by cells, so that the composition of the present invention has excellent effects of promoting collagen production and anti-aging.

[0023] Furthermore, the mass ratio of dimethyl isosorbide to inositol can also be selected as 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1 or a range between any two ratios.

[0024] Furthermore, the polypeptide composition contains, by mass percentage, 0.001-0.9% tripeptide-10 citrulline, 0.001-0.45% hexapeptide-9, 0.001-0.45% hydrolyzed sodium hyaluronate, 0.001-0.6% γ-aminobutyric acid, 0.001-1.2% decarboxylated carnosine hydrochloride, 0.001-0.4% acetyl tetrapeptide-3, 0.001-0.5% acetyl tetrapeptide-11, 0.001-0.42% palmitoyl dipeptide-7, 0.001-0.08% palmitoyl pentapeptide-4, 1-4.5% penetration enhancer and the remainder water.

[0025] On the other hand, the present invention also provides a method for preparing the composition, comprising the following steps: adding water into a container, heating, adding the remaining raw materials, stirring evenly, cooling, and sterilizing to obtain the composition.

[0026] Furthermore, the heating temperature is 30-50°C, 32-48°C, or 35-47°C.

[0027] Furthermore, the stirring speed is 20-1000 rpm, 25-800 rpm, 28-500 rpm, 30-400 rpm, 35-300 rpm or 40-200 rpm.

[0028] Furthermore, the stirring time is 0.1-10 hours, 0.1-9 hours, 0.15-7 hours, 0.2-6 hours, 0.25-5 hours, 0.3-4 hours or 0.35-1 hour.

[0029] Furthermore, the temperature is lowered to 15-28°C, 16-27°C, 17-26°C, 18-26°C, 19-25°C.

[0030] Furthermore, the present invention also provides a method for preparing the composition, comprising the following steps: adding water to a container, heating to 30-60° C., adding the remaining raw materials, stirring at a rotation speed of 20-1000 rpm for 0.1-10 hours until uniform, cooling to 15-28° C., sterilizing, and obtaining the composition.

[0031] In another aspect, the present invention also provides the use of the composition in medicine and beauty.

[0032] Furthermore, the present invention also provides use of the composition in preparing cosmetics or medicines.

[0033] Furthermore, the application includes the application of the composition in the preparation of a medicine. The medicine is used for external use on the skin.

[0034] Furthermore, the application includes using the composition in preparing cosmetics.

[0035] Furthermore, the cosmetics include facial cream, honey, lotion, facial mask, hair milk, hair spray, cream, essence, lotion, foundation, firming water, toner and the like.

[0036] Beneficial effects:

[0037] The composition of the present invention uses tripeptide-10 citrulline, hexapeptide-9, hydrolyzed sodium hyaluronate, γ-aminobutyric acid, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, palmitoyl pentapeptide-4 and other raw materials to promote the endogenous production of collagen; at the same time, the present invention uses a compound penetration enhancer to further promote the endogenous production of collagen in the skin; the composition obtained from the raw materials of the present invention can well supplement skin collagen endogenously and has an excellent anti-aging effect.

[0038] The composition obtained by the present invention has a significant effect on promoting the production of type I and type III collagen. The efficiency δ of promoting the endogenous production of type I collagen reaches 137.6%, and the efficiency ξ of promoting the endogenous production of type III collagen reaches 92.2%.

[0039] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid of the present invention is within the range of 2-4:1, and the obtained composition exhibits a better effect in promoting the production of type I collagen; γ-aminobutyric acid has an excellent effect in promoting the production of type III collagen.

[0040] The dimethyl isosorbide and inositol used in the present invention exhibit a mutually reinforcing synergistic effect in promoting the endogenous production of type I and type III collagen, and the combination of the two can enable the composition of the present invention to produce a good effect. When the mass ratio of the two is 1-2:1, the obtained composition has the best effect in promoting collagen production.

[0041] The composition of the present invention has an excellent anti-aging effect, and can reduce the proportion of skin wrinkles by 47.56% after 45 days of use, effectively giving the skin a rejuvenating effect. The combination of ingredients such as tripeptide-10 citrulline, γ-aminobutyric acid, isosorbide dimethyl ether, inositol and hexapeptide-9, hydrolyzed sodium hyaluronate, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, and palmitoyl pentapeptide-4 can give the composition of the present invention a significant anti-aging effect.

[0042] The preparation method of the composition of the invention is simple, green and environmentally friendly, and has broad application prospects. DETAILED DESCRIPTION

[0043] The present invention will be described below in conjunction with specific embodiments. These specific embodiments are used to illustrate the present invention rather than to limit the present invention.

[0044] 1. Preparation of the composition

[0045] 1. Composition 1

[0046] Raw material composition, mass percentage usage: 0.11% tripeptide-10 citrulline, 0.08% hexapeptide-9, 0.06% hydrolyzed sodium hyaluronate, 0.05% γ-aminobutyric acid, 0.04% decarboxylated carnosine hydrochloride, 0.025% acetyl tetrapeptide-3, 0.024% acetyl tetrapeptide-11, 0.018% palmitoyl dipeptide-7, 0.01% palmitoyl pentapeptide-4, 4.4% penetration enhancer and the balance water.

[0047] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 2.2:1.

[0048] The penetration enhancer is composed of isosorbide dimethyl ether and inositol in a mass ratio of 1.2:1.

[0049] Preparation method of the composition: add water into a container, heat to 42° C., add the remaining raw materials, stir at a speed of 60 rpm for 0.3 hour until uniform, cool to 25° C., sterilize, and obtain the composition 1.

[0050] 2. Composition 2

[0051] Raw material composition, mass percentage usage: 0.132% tripeptide-10 citrulline, 0.09% hexapeptide-9, 0.04% hydrolyzed sodium hyaluronate, 0.033% γ-aminobutyric acid, 0.03% decarboxylated carnosine hydrochloride, 0.035% acetyl tetrapeptide-3, 0.034% acetyl tetrapeptide-11, 0.015% palmitoyl dipeptide-7, 0.015% palmitoyl pentapeptide-4, 4.2% penetration enhancer and balance water.

[0052] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 4:1.

[0053] The penetration enhancer is composed of isosorbide dimethyl ether and inositol in a mass ratio of 2:1.

[0054] Preparation method of the composition: add water into a container, heat to 40°C, add the remaining raw materials, stir at a speed of 70 rpm for 0.4 hours until uniform, cool to 23°C, sterilize, and obtain the composition 2.

[0055] 3. Composition 3

[0056] Raw material composition, mass percentage usage: 0.06% tripeptide-10 citrulline, 0.08% hexapeptide-9, 0.06% hydrolyzed sodium hyaluronate, 0.1% γ-aminobutyric acid, 0.04% decarboxylated carnosine hydrochloride, 0.025% acetyl tetrapeptide-3, 0.024% acetyl tetrapeptide-11, 0.018% palmitoyl dipeptide-7, 0.01% palmitoyl pentapeptide-4, 4.4% penetration enhancer and the balance water.

[0057] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 0.6:1.

[0058] The penetration enhancer is composed of isosorbide dimethyl ether and inositol in a mass ratio of 1.2:1.

[0059] Preparation method of the composition: add water into a container, heat to 42° C., add the remaining raw materials, stir at a speed of 60 rpm for 0.3 hour until uniform, cool to 25° C., sterilize, and obtain the composition 3.

[0060] 4. Composition 4

[0061] Raw material composition, mass percentage usage: 0.095% tripeptide-10 citrulline, 0.08% hexapeptide-9, 0.06% hydrolyzed sodium hyaluronate, 0.065% γ-aminobutyric acid, 0.04% decarboxylated carnosine hydrochloride, 0.025% acetyl tetrapeptide-3, 0.024% acetyl tetrapeptide-11, 0.018% palmitoyl dipeptide-7, 0.01% palmitoyl pentapeptide-4, 4.4% penetration enhancer and balance water.

[0062] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 1.46:1.

[0063] The penetration enhancer is composed of isosorbide dimethyl ether and inositol in a mass ratio of 1.2:1.

[0064] Preparation method of the composition: add water into a container, heat to 42°C, add the remaining raw materials, stir at a speed of 60 rpm for 0.3 hour until uniform, cool to 25°C, sterilize, and obtain the composition 4.

[0065] 5. Composition 5

[0066] Raw material composition, mass percentage usage: 0.14% tripeptide-10 citrulline, 0.08% hexapeptide-9, 0.06% hydrolyzed sodium hyaluronate, 0.02% γ-aminobutyric acid, 0.04% decarboxylated carnosine hydrochloride, 0.025% acetyl tetrapeptide-3, 0.024% acetyl tetrapeptide-11, 0.018% palmitoyl dipeptide-7, 0.01% palmitoyl pentapeptide-4, 4.4% penetration enhancer and the balance water.

[0067] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 7:1.

[0068] The penetration enhancer is composed of isosorbide dimethyl ether and inositol in a mass ratio of 1.2:1.

[0069] Preparation method of the composition: add water into a container, heat to 42° C., add the remaining raw materials, stir at a speed of 60 rpm for 0.3 hour until uniform, cool to 25° C., sterilize, and obtain the composition 5.

[0070] 6. Composition 6

[0071] Raw material composition, mass percentage usage: 0.15% tripeptide-10 citrulline, 0.08% hexapeptide-9, 0.06% hydrolyzed sodium hyaluronate, 0.01% γ-aminobutyric acid, 0.04% decarboxylated carnosine hydrochloride, 0.025% acetyl tetrapeptide-3, 0.024% acetyl tetrapeptide-11, 0.018% palmitoyl dipeptide-7, 0.01% palmitoyl pentapeptide-4, 4.4% penetration enhancer and the balance water.

[0072] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 15:1.

[0073] The penetration enhancer is composed of isosorbide dimethyl ether and inositol in a mass ratio of 1.2:1.

[0074] Preparation method of the composition: add water into a container, heat to 42°C, add the remaining raw materials, stir at a speed of 60 rpm for 0.3 hour until uniform, cool to 25°C, sterilize, and obtain the composition 6.

[0075] 7. Composition 7

[0076] Raw material composition, mass percentage usage: 0.11% tripeptide-10 citrulline, 0.08% hexapeptide-9, 0.06% hydrolyzed sodium hyaluronate, 0.05% γ-aminobutyric acid, 0.04% decarboxylated carnosine hydrochloride, 0.025% acetyl tetrapeptide-3, 0.024% acetyl tetrapeptide-11, 0.018% palmitoyl dipeptide-7, 0.01% palmitoyl pentapeptide-4, 4.4% penetration enhancer and the balance water.

[0077] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 2.2:1.

[0078] The penetration enhancer is inositol.

[0079] Preparation method of the composition: add water into a container, heat to 42°C, add the remaining raw materials, stir at a speed of 60 rpm for 0.3 hour until uniform, cool to 25°C, sterilize, and obtain the composition 7.

[0080] 8. Composition 8

[0081] Raw material composition, mass percentage usage: 0.11% tripeptide-10 citrulline, 0.08% hexapeptide-9, 0.06% hydrolyzed sodium hyaluronate, 0.05% γ-aminobutyric acid, 0.04% decarboxylated carnosine hydrochloride, 0.025% acetyl tetrapeptide-3, 0.024% acetyl tetrapeptide-11, 0.018% palmitoyl dipeptide-7, 0.01% palmitoyl pentapeptide-4, 4.4% penetration enhancer and the balance water.

[0082] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 2.2:1.

[0083] The penetration enhancer is dimethyl isosorbide.

[0084] Preparation method of the composition: add water into a container, heat to 42°C, add the remaining raw materials, stir at a speed of 60 rpm for 0.3 hour until uniform, cool to 25°C, sterilize, and obtain the composition 8.

[0085] 9. Composition 9

[0086] Raw material composition, mass percentage usage: 0.11% tripeptide-10 citrulline, 0.08% hexapeptide-9, 0.06% hydrolyzed sodium hyaluronate, 0.05% γ-aminobutyric acid, 0.04% decarboxylated carnosine hydrochloride, 0.025% acetyl tetrapeptide-3, 0.024% acetyl tetrapeptide-11, 0.018% palmitoyl dipeptide-7, 0.01% palmitoyl pentapeptide-4, 4.4% penetration enhancer and the balance water.

[0087] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 2.2:1.

[0088] The penetration enhancer is composed of isosorbide dimethyl ether and inositol in a mass ratio of 0.3:1.

[0089] Preparation method of the composition: add water into a container, heat to 42°C, add the remaining raw materials, stir at a speed of 60 rpm for 0.3 hour until uniform, cool to 25°C, sterilize, and obtain the composition 9.

[0090] 10. Composition 10

[0091] Raw material composition, mass percentage usage: 0.11% tripeptide-10 citrulline, 0.08% hexapeptide-9, 0.06% hydrolyzed sodium hyaluronate, 0.05% γ-aminobutyric acid, 0.04% decarboxylated carnosine hydrochloride, 0.025% acetyl tetrapeptide-3, 0.024% acetyl tetrapeptide-11, 0.018% palmitoyl dipeptide-7, 0.01% palmitoyl pentapeptide-4, 4.4% penetration enhancer and the balance water.

[0092] The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 2.2:1.

[0093] The penetration enhancer is composed of isosorbide dimethyl ether and inositol in a mass ratio of 3.5:1.

[0094] The preparation method of the composition is as follows: water is added into a container, heated to 42° C., the remaining raw materials are added, stirred at a rotation speed of 60 rpm for 0.3 hour until uniform, cooled to 25° C., and sterilized to obtain the composition 10.

[0095] 2. Performance Test

[0096] The above polypeptide compositions 1-10 were subjected to the following performance tests.

[0097] 1. Test on the effect of promoting endogenous production of type I and type III collagen

[0098] Test as follows:

[0099] Blank control group: HSF dermal fibroblasts were selected as the cell model, with 1.8×10 5 The cells were inoculated at a density of 100 cells / well in a 6-well plate and cultured at 5.2% CO2 and 37°C for 1 day. When the cell plating rate in the 6-well plate reached about 50%, about 2 mL of DMEM culture medium was added to each well to obtain a blank control group.

[0100] The pretreatment of the composition 1-10 groups was carried out in the same manner as the blank control group, specifically as follows: dermal fibroblasts HSF were selected as the cell model, and 1.8×10 5 The cells were inoculated at a density of cells / well in a 6-well plate and cultured at 5.2% CO2 and 37°C for 1 day. When the cell plating rate in the 6-well plate reached about 50%, about 2.2 mL of DMEM culture medium was added to each well, and the above-mentioned composition 1-10 was added to each well and the mass concentration of the composition in the culture medium was controlled to be 20 ppm, to obtain a group of composition 1-10.

[0101] After adding culture medium (and composition 1-10) to the blank control group and composition groups 1-10, the well plates of each group were placed in an incubator and incubated in an environment of 5.2% CO2 and 37°C for 60 h, the culture medium was aspirated, and the cells were washed twice with 10 times the volume of PBS, and the cells were digested with 0.25wt% trypsin for 0.3 h, centrifuged at 3500 rpm for 4 min, and the supernatant was collected in a test tube. The cells were washed once with 10 times the volume of PBS, and the supernatant was collected in an EP tube by centrifugation again to obtain the objects to be detected for the blank control group and composition groups 1-10.

[0102] The ELISA test kit was used to detect the type I and type III collagen contents in the object to be detected. Then the efficiency of promoting the endogenous generation of collagen was calculated, wherein the efficiency of promoting the endogenous generation of type I collagen δ=100%×(the content of type I collagen in the object to be detected in the composition group-the content of type I collagen in the object to be detected in the blank control group) / the content of type I collagen in the object to be detected in the blank control group, and the efficiency of promoting the endogenous generation of type III collagen ξ=100%×(the content of type III collagen in the object to be detected in the composition group-the content of type III collagen in the object to be detected in the blank control group) / the content of type III collagen in the object to be detected in the blank control group. The detailed results of promoting the endogenous generation of type I and type III collagen are shown in Tables 1 and 2.

[0103] Table 1: Effects of promoting endogenous production of type I collagen

[0104]

[0105] Table 2: Effect of promoting endogenous production of type III collagen

[0106]

[0107] From the test results in Table 1-2, it can be seen that the composition of the present invention uses tripeptide-10 citrulline, hexapeptide-9, hydrolyzed sodium hyaluronate, γ-aminobutyric acid, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, and palmitoyl pentapeptide-4 to promote the endogenous production of collagen; at the same time, the present invention uses a compound penetration enhancer to further promote the endogenous production of collagen in the skin; the composition obtained from the raw materials of the present invention has a good effect of endogenously supplementing skin collagen. The efficiency δ of the composition obtained by the present invention in promoting the endogenous production of type I collagen reached 137.6%, and the efficiency ξ of promoting the endogenous production of type III collagen reached 92.2%.

[0108] The mass ratio of tripeptide-10 citrulline and γ-aminobutyric acid in composition 1 of the present invention is within the range of 2-4:1, and the ratio of compositions 3-6 is outside the range of 2-4:1; and the comparison results of the test in Table 1 show that the mass ratio of tripeptide-10 citrulline and γ-aminobutyric acid is within the range of 2-4:1, and the obtained composition exhibits a better effect in promoting the production of type I collagen; and the test results of compositions 3-4 and 1 in Table 2 show that γ-aminobutyric acid has an excellent effect in promoting the production of type III collagen.

[0109] From the test results of compositions 1, 7 and 8 in Tables 1 and 2, it can be seen that dimethyl isosorbide and inositol used in the present invention exhibit a mutually reinforcing synergistic effect in promoting the endogenous production of type I and type III collagen, and the combination of the two can make the composition of the present invention produce a good effect. When the mass ratio of the two is 1-2:1, the obtained composition has the best effect in promoting collagen production.

[0110] 2. Anti-aging effect test

[0111] The anti-aging effect of the composition is characterized by the degree of reduction of wrinkles at the corners of the eyes after using the composition.

[0112] Steps for using the test sample: recruit volunteers aged 40-55 years old and divide them into 10 groups, with 6 people in each group, 3 males and 3 females; before using composition 1-10, the volunteers in each group clean the skin near the eyes with clean water, and then apply the above composition 1-10 to the skin of the corners of both eyes once every night, using 0.3g each time, and use it continuously for 45 days.

[0113] Test method: Before the test, the skin of the eye corners of the volunteers was cleaned with clean water; then the wrinkles of the skin of the eye corners of the eyes before and 45 days after using the composition were analyzed using a VISIA facial image analyzer, and then the reduction of the skin wrinkles of the eye corners of the eyes before and after using the composition was analyzed to obtain the wrinkle reduction rate A value / %; wherein the 45-day skin wrinkle reduction rate A value of the eye corners of the eyes is the ratio of the reduction amount of the skin wrinkles of the eye corners of the eyes before using the composition on the 45th day to the amount of the skin wrinkles of the eye corners of the eyes before using the composition. The specific results of the anti-aging effect test are shown in Table 3.

[0114] Table 3: Anti-aging effect test

[0115]

[0116] As can be seen from the test results in Table 3, the composition of the present invention has an excellent anti-aging effect, and can reduce the proportion of skin wrinkles by 47.56% after 45 days of use, and can effectively give the skin a rejuvenating effect. As can be seen from the test results of compositions 1, 3-10, the combination of tripeptide-10 citrulline, γ-aminobutyric acid, isosorbide dimethyl ether, inositol and hexapeptide-9, hydrolyzed sodium hyaluronate, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, palmitoyl pentapeptide-4 and other ingredients can give the composition of the present invention a significant anti-aging effect.

[0117] 3. Application Examples

[0118] 1. Facial mask

[0119] The raw materials and their dosages are as follows: 5% composition 1, 2% glycerol, 2.5% 1,3-butylene glycol, 1.5% beta-glucan, 3% betaine, 0.5% cucumber extract, 0.8% licorice extract, 0.4% Centella asiatica extract, 0.3% meadowfoam seed oil, 0.5% jojoba seed oil, 0.2% squalane, 0.2% acrylic acid (esters) / C10-30 alkyl acrylate cross-linked polymer, 0.4% hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer, 0.2% Tween-60, 0.5% polyglycerol-10 caprate, 2% 1,2-pentanediol, 0.2% arginine, 0.1% EDTA-2Na and the balance water; measured in mass percentage.

[0120] Preparation method: add water to a container, add acrylic acid (ester) / C10-30 alkyl acrylate cross-linked polymer, hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer, stir and heat to 85°C at a speed of 100 rpm, stir for 0.5 hour until uniform after complete dissolution; then add glycerol, 1,3-butylene glycol, β-glucan, betaine, Tween-60, polyglycerol-10 caprate, and continue stirring for 0.2 hour; then add white meadowfoam seed oil, jojoba seed oil, squalane, stir and homogenize for 0.1 hour, cool to 45°C, add cucumber extract, licorice extract, Centella asiatica extract, 1,2-pentanediol, arginine, EDTA-2Na and composition 1 and other remaining raw materials, continue stirring for 0.3 hour, let stand, and obtain the facial mask.

[0121] 2. Essence

[0122] The raw materials and their dosage are as follows: 7% composition 1, 3% glycerol, 3.5% 1,3-butylene glycol, 2% beta-glucan, 0.5% rye seed extract, 0.7% rhodiola rosea extract, 0.5% tea extract, 0.1% camellia oil, 0.8% jojoba seed oil, 0.5% caprylic / capric triglyceride, 1.2% hydrogenated lecithin, 0.5% laureth-5, 2% 1,2-pentanediol, 0.1% EDTA-2Na and the balance water; measured in mass percentage.

[0123] Preparation method: add water into a container, heat it to 85°C, add glycerol, 1,3-butylene glycol, and β-glucan, stir at a speed of 100 rpm for 0.4 hour and stir evenly; then add hydrogenated lecithin and laureth-5, continue stirring for 0.2 hour; then add camellia oil, jojoba seed oil, caprylic / capric triglyceride, stir and homogenize for 0.3 hour, cool to 45°C, add composition 1, 1,2-pentanediol, EDTA-2Na and other remaining raw materials, continue stirring for 0.2 hour, let stand, and obtain the essence.

[0124] The composition of the present invention uses raw materials such as tripeptide-10 citrulline, hexapeptide-9, hydrolyzed sodium hyaluronate, γ-aminobutyric acid, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, palmitoyl pentapeptide-4, etc. Each raw material plays a role and cooperates with each other to promote the coordinated expression of skin type I, type III, type IV, type VII, type XVII collagen, fibronectin, elastin and glycosaminoglycans; prevent collagen cross-linking, maintain the triple helix structure, and inhibit collagen degradation. When applied to skin medicine and cosmetology, it can accelerate the regeneration of structural proteins such as skin collagen, repair skin damage, reduce wrinkles such as crow's feet, tear grooves, nasolabial folds, and under-eye wrinkles, solve the skin aging problem caused by the loss of structural proteins such as collagen, and improve skin elasticity and firmness.

[0125] The present invention obtains a polypeptide composition by compounding a specific ratio of cell active peptides and a component that promotes penetration. Compared with exogenous collagen supplementation, it can be effectively absorbed through the skin, enhance the ability of autologous collagen regeneration, better regulate the extracellular matrix, and is safer and more effective. By endogenously stimulating the regeneration of structural proteins such as skin collagen, the problem of skin sagging and aging caused by the loss of structural proteins such as skin collagen is completely solved.

[0126] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A cell active peptide composition for promoting the regeneration of multi-type collagen structural protein, characterized in that: Measured in percentage by mass, it contains: 0.0001-1% tripeptide-10 citrulline, 0.0001-0.5% hexapeptide-9, 0.0001-0.5% hydrolyzed sodium hyaluronate, 0.0001-2% γ-aminobutyric acid, 0.0001-1.5% decarboxylated carnosine hydrochloride, 0.0001-1% acetyl tetrapeptide-3, 0.0001-1% acetyl tetrapeptide-11, 0.0001-0.5% palmitoyl dipeptide-7, 0.0001-0.1% palmitoyl pentapeptide-4, 0.5-5% penetration enhancer and the remainder water.

2. The cell active peptide composition according to claim 1, characterized in that The mass ratio of the tripeptide-10 citrulline to γ-aminobutyric acid is 2-4:

1.

3. The cell active peptide composition according to claim 1, characterized in that: The penetration enhancer is dimethyl isosorbide and / or inositol.

4. The cell active peptide composition according to claim 1, characterized in that The penetration enhancer is a combination of isosorbide dimethyl ether and inositol in a mass ratio of 1-2:

1.

5. The cell active peptide composition according to claim 1, characterized in that: The dosage of the tripeptide-10 citrulline is 0.001-0.9%.

6. The cell active peptide composition according to claim 1, characterized in that: The molecular weight of the hydrolyzed sodium hyaluronate is less than 1000Da.

7. The cell active peptide composition according to claim 1, characterized in that: The dosage of the γ-aminobutyric acid is 0.001-0.6%.

8. The cell active peptide composition according to claim 1, characterized in that: Calculated by mass percentage, it contains: 0.001-0.9% tripeptide-10 citrulline, 0.001-0.45% hexapeptide-9, 0.001-0.45% hydrolyzed sodium hyaluronate, 0.001-0.6% γ-aminobutyric acid, 0.001-1.2% decarboxylated carnosine hydrochloride, 0.001-0.4% acetyl tetrapeptide-3, 0.001-0.5% acetyl tetrapeptide-11, 0.001-0.42% palmitoyl dipeptide-7, 0.001-0.08% palmitoyl pentapeptide-4, 1-4.5% penetration enhancer and the balance water.

9. A method for preparing the cell active peptide composition according to any one of claims 1 to 8, characterized in that: The method comprises the following steps: adding water into a container, heating, adding remaining raw materials, stirring evenly, cooling, and sterilizing to obtain the cell active peptide composition.

10. Use of the cell active peptide composition according to claims 1 to 8 in the preparation of cosmetics or medicines.

Images