Cell active peptide composition for promoting regeneration of multitype collagen structural protein, preparation method thereof, and application in medicine and cosmetology

Abstract

The present invention relates to the fields of medicine and cosmetology, and discloses a cell active peptide composition that promotes the regeneration of multi-type collagen structural proteins, its preparation method, and its application in medicine and cosmetology. The composition 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, and palmitoyl pentapeptide-4. The composition can 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 triple helix structure, and inhibit collagen degradation. Applied to dermatology 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 groove lines, nasolabial folds, and under-eye lines, solve the skin aging problem caused by the loss of structural proteins such as collagen, and improve skin elasticity and firmness.

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 multitype collagen structural protein, a preparation method thereof, and application in medicine and cosmetology. Background Art

[0002] Collagen is the primary component of the skin's extracellular matrix and plays a vital role in maintaining the skin's plumpness, elasticity, and overall youthfulness. However, with the passage of time, the collagen content in the skin gradually decreases. This natural physiological process can lead to signs of skin aging, such as sagging and wrinkles. In addition, reactive oxygen species (ROS) are one of the key factors that accelerate collagen loss and promote skin aging. During the photoaging process, ultraviolet radiation stimulates the production of ROS in the skin, which in turn accelerates collagen degradation, disrupts 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, comprising 60% to 80% of the total collagen content in the dermis and providing the skin's primary strength and support. Close behind is type III collagen, which accounts for 15% to 25% of the dermis' collagen content and plays a key role in the skin's flexibility and repair capacity. Together, these two types of collagen form the foundation of the structural fibers in the dermis, supporting the skin's structural integrity.

[0004] There are multiple ways to supplement collagen: exogenous supplementation, reducing collagen degradation, and endogenous collagen regeneration. Exogenous supplementation mainly involves injections of recombinant humanized collagen or animal-derived collagen; reducing collagen degradation mainly involves inhibiting the activity of metalloproteinases, and preventing protein cross-linking through antioxidant and anti-glycation effects; 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, addressing the root cause of structural protein loss.

[0005] Although collagen supplementation has potential benefits for combating skin aging, current collagen supplementation methods on the market still face some challenges. For example, collagen derived from animals may pose risks of immunogenicity and disease transmission. While recombinant humanized collagen has advantages in reducing immunogenicity, it still faces many issues 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, making it difficult for them to penetrate the epidermis and reach the dermis, and therefore 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 response to the above technical problems, the inventors proposed the technical solution of the present invention, which includes a cell active peptide composition that promotes 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 promotes the production of type I and type IV collagen. Palmitoyl dipeptide-7 inhibits the activity of metalloproteinases, increasing the expression of structural proteins and hyaluronic acid in the dermis and basement membrane. Tripeptide-10 citrulline ensures uniform fiber diameter, allowing collagen fibers to cohere better, thereby increasing skin softness. Acetyl tetrapeptide-3 acts on fibroblasts, accelerating the synthesis of extracellular matrix proteins such as laminin, collagen III, and collagen VII. Acetyl tetrapeptide-11 reshapes and strengthens skin structure by simultaneously stimulating the synthesis of syndecan-1 and type XVII collagen. Palmitoyl pentapeptide-4, a sequence at positions 212-216 of type I collagen, increases the expression of type III collagen in fibroblasts. Decarboxylated carnosine hydrochloride binds to glycated proteins, preventing further cross-linking of glycated proteins, effectively inhibiting advanced glycation end products (AGEs), and protecting collagen. Gamma-aminobutyric acid: It promotes elastin synthesis and elastic fiber formation 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 carbohydrate, 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 an 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 weight 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 balance 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 1000 Da.

[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] Furthermore, 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. Furthermore, the penetration enhancer is selected to be a combination of dimethyl isosorbide and inositol in a mass ratio of 1-2:1. Under such conditions, 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 collagen production promotion and anti-aging effects.

[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 weight percentage, the following: 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.

[0025] On the other hand, the present invention also provides a method for preparing the composition, comprising the steps of 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, or 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 speed of 20-1000 rpm for 0.1-10 hours until uniform, cooling to 15-28° C., and sterilizing to obtain the composition.

[0031] On the other hand, 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 using the composition in preparing a medicine 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, etc.

[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 gamma-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; gamma-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 synergistic effect in promoting the endogenous production of types I and III collagen. Their combination produces excellent results. The optimal collagen production-promoting effect is achieved when the mass ratio of the two is 1-2:1.

[0041] The composition of the present invention has excellent anti-aging effects, reducing the proportion of skin wrinkles by 47.56% after 45 days of use, effectively imparting a youthful effect to the skin. The combination of ingredients such as tripeptide-10 citrulline, gamma-aminobutyric acid, dimethyl isosorbide, inositol, hexapeptide-9, hydrolyzed sodium hyaluronate, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, and palmitoyl pentapeptide-4 imparts significant anti-aging benefits to the composition of the present invention.

[0042] The preparation method of the composition of the present 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 gamma-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 60 rpm for 0.3 hour until uniform, cool to 25° C., and sterilize to 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 gamma-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 70 rpm for 0.4 hour until uniform, cool to 23°C, and sterilize to 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 balance water.

[0057] The mass ratio of the tripeptide-10 citrulline to gamma-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 60 rpm for 0.3 hour until uniform, cool to 25° C., and sterilize to 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 gamma-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 60 rpm for 0.3 hour until uniform, cool to 25° C., and sterilize to 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 gamma-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 60 rpm for 0.3 hour until uniform, cool to 25° C., and sterilize to 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 balance water.

[0072] The mass ratio of the tripeptide-10 citrulline to gamma-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, and sterilize to 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 gamma-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 60 rpm for 0.3 hour until uniform, cool to 25° C., and sterilize to 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 gamma-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 60 rpm for 0.3 hour until uniform, cool to 25°C, and sterilize to 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 gamma-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 to a container, heat to 42° C., add the remaining raw materials, stir at 60 rpm for 0.3 hour until uniform, cool to 25° C., and sterilize to 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 gamma-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 to a container, heated to 42° C., the remaining raw materials are added, and the mixture is stirred at 60 rpm for 0.3 hours until uniform, cooled to 25° C., and sterilized to obtain the composition 10.

[0095] 2. Performance Testing

[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: dermal fibroblasts HSF were selected as the cell model, with 1.8×10 5 The cells were seeded 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 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 seeded at a density of 100 cells / well in a 6-well plate and incubated at 5.2% CO2 and 37°C for 1 day. When the cell plating rate in the 6-well plate reached approximately 50%, approximately 2.2 mL of DMEM medium was added to each well, and the above-mentioned compositions 1-10 were added to each well while controlling the mass concentration of the composition in the culture medium to 20 ppm, to obtain groups of compositions 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 a 5.2% CO2, 37°C environment for 60 hours. The culture medium was aspirated and washed twice with 10 times the volume of PBS. The cells were digested with 0.25wt% trypsin for 0.3 hours, centrifuged at 3500 rpm for 4 minutes, 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 by centrifugation again in an EP tube to obtain the objects to be detected for the blank control group and composition groups 1-10.

[0102] The type I and type III collagen levels in the test subjects were measured using an ELISA test kit. The efficiency of promoting endogenous collagen production was then calculated: the efficiency of promoting endogenous type I collagen production (δ) = 100% × (type I collagen content in the test subject in the composition group - type I collagen content in the test subject in the blank control group) / type I collagen content in the test subject in the blank control group; the efficiency of promoting endogenous type III collagen production (ξ) = 100% × (type III collagen content in the test subject in the composition group - type III collagen content in the test subject in the blank control group) / type III collagen content in the test subject in the blank control group. Detailed results of the effects of promoting endogenous type I and type III collagen production are shown in Tables 1 and 2.

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

[0104]

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

[0106]

[0107] As shown in the test results in Table 1-2, 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 collagen production. Furthermore, the present invention uses a compounded penetration enhancer to further promote endogenous collagen production within the skin. The composition obtained from the raw materials of the present invention has a good effect of endogenously replenishing 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 resulting 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] The test results for compositions 1, 7, and 8 in Tables 1 and 2 demonstrate that dimethyl isosorbide and inositol exhibit a synergistic effect in promoting the endogenous production of types I and III collagen. Their combination yields excellent results. The optimal collagen-promoting effect was achieved when the mass ratio of dimethyl isosorbide and inositol was 1-2:1.

[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] The following steps were used to test the sample: volunteers aged 40-55 were recruited and divided into 10 groups, with 6 people in each group, 3 men and 3 women. Before using Composition 1-10, the volunteers in each group washed the skin near the eyes with clean water, and then applied 0.3g of Composition 1-10 to the skin of the corners of both eyes once every night for 45 consecutive days.

[0113] Testing method: Before the test, the skin at the canthus of both eyes of the volunteer was cleaned with clean water; then, a VISIA facial image analyzer was used to analyze the wrinkles of the skin at the canthus of both eyes before and 45 days after using the composition, and then the reduction of the skin wrinkles at the canthus of both eyes before and after using the composition was analyzed to obtain a wrinkle reduction rate A value / %. The 45-day skin wrinkle reduction rate A value is the ratio of the amount of skin wrinkles at the canthus of both eyes reduced on the 45th day compared to before using the composition to the amount of skin wrinkles at the canthus of both 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. After 45 days of use, it can reduce the proportion of skin wrinkles by 47.56%, and can effectively give the skin a youthful effect. The test results of compositions 1, 3-10 show that the combination of tripeptide-10 citrulline, γ-aminobutyric acid, dimethyl isosorbide, 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 dosage are as follows: 5% composition 1, 2% glycerin, 2.5% 1,3-butylene glycol, 1.5% betaine, 3% cucumber extract, 0.5% licorice extract, 0.4% Centella asiatica extract, 0.3% meadowfoam seed oil, 0.5% jojoba seed oil, 0.2% squalane, 0.2% acrylates / C10-30 alkyl acrylate crosspolymer, 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; the dosage is measured in mass percentage.

[0120] Preparation method: add water to a container, add acrylic acid (ester) / C10-30 alkyl acrylate cross-polymer and hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer, and stir and heat to 85°C at a speed of 100 rpm. After complete dissolution, stir for 0.5 hour until uniform; then add glycerin, 1,3-butylene glycol, β-glucan, betaine, Tween-60, and polyglycerol-10 caprate, and continue stirring for 0.2 hour; then add white meadowfoam seed oil, jojoba seed oil, and 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, and let stand to obtain the facial mask.

[0121] 2. Essence

[0122] The raw materials and their usage 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; the amounts are measured in percentage by mass.

[0123] Preparation method: add water to a container, heat to 85°C, add glycerol, 1,3-butylene glycol, and β-glucan, and stir at 100 rpm for 0.4 hour to mix evenly; then add hydrogenated lecithin and laureth-5, and continue stirring for 0.2 hour; then add camellia oil, jojoba seed oil, and 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, and let stand to obtain the essence.

[0124] The composition of the present invention uses ingredients such as tripeptide-10 citrulline, hexapeptide-9, hydrolyzed sodium hyaluronate, gamma-aminobutyric acid, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, and palmitoyl pentapeptide-4. Each ingredient works together to promote the coordinated expression of skin types I, III, IV, VII, and XVII collagen, fibronectin, elastin, and glycosaminoglycans; prevent collagen cross-linking, maintain the triple helix structure, and inhibit collagen degradation. Applied in dermatology and cosmetics, it can accelerate the regeneration of structural proteins such as collagen in the skin, repair skin damage, reduce wrinkles such as crow's feet, tear troughs, nasolabial folds, and under-eye lines, address skin aging issues caused by the loss of structural proteins such as collagen, and improve skin elasticity and firmness.

[0125] This invention combines a specific ratio of active peptides with a permeation-promoting ingredient to create a peptide composition. Compared to exogenous collagen supplementation, this peptide composition is more effectively absorbed through the skin, enhancing autologous collagen regeneration and better regulating the extracellular matrix, making it safer and more effective. By endogenously stimulating the regeneration of structural proteins like collagen in the skin, it completely resolves the problem of sagging and aging skin caused by the loss of these structural proteins.

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

Claims

1. A cell active peptide composition for promoting the regeneration of multi-type collagen structural protein, characterized in that: Calculated by mass percentage, the composition is: 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; the mass ratio of tripeptide-10 citrulline to γ-aminobutyric acid is 2-4:1; the penetration enhancer is a combination of dimethyl isosorbide and inositol in a mass ratio of 1-2:

1.

2. The cell active peptide composition according to claim 1, wherein The molecular weight of the hydrolyzed sodium hyaluronate is less than 1000 Da.

3. A method for preparing the cell active peptide composition according to any one of claims 1 to 2, characterized in that: The method comprises the following steps: adding water into a container, heating, adding the remaining raw materials, stirring evenly, cooling, and sterilizing to obtain the cell active peptide composition.

4. Use of the cell active peptide composition according to any one of claims 1 to 2 in the preparation of cosmetics or medicines.

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