Full-layer anti-aging recombinant collagen filling powder as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of skin care products, and relates to full-layer anti-aging recombinant collagen filling powder as well as a preparation method and application thereof. The recombinant collagen filling powder is prepared from recombinant I type collagen, recombinant III type collagen, recombinant IV type collagen, recombinant VI type collagen, recombinant XVII type collagen, superoxide dismutase, a bifidus yeast lysate, trehalose, silica, silica dimethyl silylate and a polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and the recombinant collagen filling powder is prepared from the following raw materials: recombinant I type collagen, recombinant III type collagen, recombinant IV type collagen, recombinant VI type collagen, recombinant XVII type collagen, superoxide dismutase, bifidus yeast lysate, trehalose, silica, silica dimethyl silylate and a polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer. The skin care product is prepared from glycerin, xanthan gum, sodium hyaluronate, dimeticone, phenoxyethanol and ethylhexylglycerin. The recombinant collagen filling powder provided by the invention is convenient to use, can obviously improve the skin moisturizing capacity, improve the compactness and repair wrinkles, has a good anti-aging effect and a good repair effect, and has important practical significance and market value.

Description

Technical Field

[0001] The present invention belongs to the technical field of skin care products, and specifically relates to a recombinant collagen filling powder for full-layer anti-aging, and its preparation method and application. Background Art

[0002] Skin aging is a complex biological process, and the skin structure and function gradually deteriorate over time. During the process of skin aging, collagen gradually loses, resulting in a series of problems such as wrinkles, skin laxity, eye bags, nasolabial folds and tear troughs. In order to achieve skin repair and rejuvenation, those skilled in the art use technical means to add collagen to skin care products or medical beauty drugs, and by supplementing collagen to the skin, improve skin firmness and promote skin repair, so as to achieve the effect of skin anti-aging. As a product of genetic engineering technology, recombinant collagen has become an important choice for collagen anti-aging products due to its high safety and other reasons. Among them, recombinant type I collagen has good support and elasticity, can stabilize water molecules and slow down their evaporation rate, stimulate cells to produce new collagen, inhibit the generation of melanin and fill the gaps of collagen lost in the dermis at the same time; recombinant type III collagen is located between the epidermis and the dermis in the skin, and is called "baby collagen", which is the key structure to support the epidermis; recombinant type IV collagen is distributed in connective tissues, can tighten the skin and repair the barrier; recombinant type VI collagen can interact with type I and type III collagens and participate in maintaining the structure and function of connective tissues. Recombinant type XVII collagen is located between the epidermis and the dermis. As a transmembrane protein on hemidesmosomes, it is responsible for tightly binding the epidermis to the dermis, ensuring the integrity of the skin structure, and at the same time regulating skin homeostasis, aging and wound repair. Some types of recombinant collagen have been used as important components in anti-aging products. For example, Chinese Patent CN118384055A discloses a highly permeable recombinant collagen liposome and its preparation method and application, which has the characteristics of high permeability, high bioavailability, good stability and convenient application, and can significantly improve the application effect of recombinant collagen.

[0003] However, how to select and scientifically proportion recombinant collagen to make the product have better performance effects needs further exploration, and it is also one of the key issues studied in the present invention; at the same time, the main application dosage forms of conventional recombinant collagen in the field of skin care products are solution state or freeze-dried powder state. The long-term stability of recombinant collagen in solution state is easily affected by factors such as environmental temperature, affecting the activity and efficacy of the product. Collagen freeze-dried powder often needs to be reconstituted with a solvent when used, and the operation is cumbersome. Therefore, providing a recombinant collagen anti-aging product with simple use, stable performance and good anti-aging performance has important market value and practical significance. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a recombinant collagen filling powder for full-layer anti-aging, its preparation method and application.

[0005] To solve the above problems, the technical solutions adopted by the present invention are as follows: Technical theme one: The present invention provides a recombinant collagen filling powder for full-layer anti-aging, containing the following components: Recombinant type I collagen with a mass fraction of 0.6% - 0.7%; Recombinant type III collagen with a mass fraction of 0.08% - 0.16%; Recombinant type IV collagen with a mass fraction of 0.008% - 0.012%; Recombinant type VI collagen with a mass fraction of 0.008% - 0.012%; Recombinant type XVII collagen with a mass fraction of 0.02% - 0.04%; Superoxide dismutase with a mass fraction of 0.001% - 0.01%; Lyophilized bifidobacterium lysate with a mass fraction of 0.1% - 2%; Trehalose with a mass fraction of 1% - 5%; Silica with a mass fraction of 10% - 20%; Dimethylsilylated silica with a mass fraction of 5% - 10%; Polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer with a mass fraction of 3% - 8%; Glycerol with a mass fraction of 8% - 20%; Xanthan gum with a mass fraction of 1% - 3%; Sodium hyaluronate with a mass fraction of 0.02% - 0.08%; Dimethicone with a mass fraction of 1% - 5%; Phenoxyethanol with a mass fraction of 0.3% - 0.5%; Ethylhexylglycerin with a mass fraction of 0.1% - 1%; And the balance of water.

[0006] As a further embodiment of the present invention, it contains the following components: Recombinant type I collagen with a mass fraction of 0.6% - 0.7%; Recombinant type III collagen with a mass fraction of 0.08% - 0.15%; Recombinant type IV collagen with a mass fraction of 0.008% - 0.012%; Recombinant type VI collagen with a mass fraction of 0.008% - 0.012%; Recombinant type XVII collagen with a mass fraction of 0.02% to 0.03%; Superoxide dismutase with a mass fraction of 0.003% to 0.008%; 0.8% to 1.2% mass fraction of bifid yeast lysate; Trehalose with a mass fraction of 2% to 4%; Silica with a mass fraction of 13% to 18%; 5% to 10% by mass of silica dimethyl silylate; 3% to 8% by mass of polydimethylsiloxane / vinyl polydimethylsiloxane crosspolymer; Glycerol with a mass fraction of 12% to 16%; Xanthan gum with a mass fraction of 1% to 3%; Sodium hyaluronate with a mass fraction of 0.03% to 0.08%; Dimethicone with a mass fraction of 1% to 5%; Phenoxyethanol with a mass fraction of 0.3% to 0.5%; Ethylhexylglycerin with a mass fraction of 0.3% to 0.8%; and the remaining water.

[0007] As a further embodiment of the present invention, the invention comprises the following ingredients: Recombinant type I collagen with a mass fraction of 0.64%; 0.12% recombinant type III collagen; 0.008% recombinant type IV collagen; 0.008% recombinant type VI collagen; 0.024% recombinant type XVII collagen; Superoxide dismutase with a mass fraction of 0.005%; 1.0% mass fraction of bifid yeast lysate; Trehalose with a mass fraction of 3.0%; 15.0% by mass of silica; 8.0% by mass of silica dimethyl silylate; 5.0% by mass of dimethicone / vinyl dimethicone crosspolymer; 14% glycerol by mass; 2.0% xanthan gum by mass; 0.05% sodium hyaluronate by mass; 3.0% dimethicone by mass; Phenoxyethanol at a mass fraction of 0.4%; Ethylhexylglycerin at a mass fraction of 0.50%; And the balance is water.

[0008] As a further embodiment of the present invention, the molecular weight of the recombinant type I collagen is 10 kDa to 90 kDa; the molecular weight of the recombinant type III collagen is 10 kDa to 90 kDa; the molecular weight of the recombinant type IV collagen is 10 kDa to 90 kDa; the molecular weight of the recombinant type VI collagen is 10 kDa to 90 kDa; the molecular weight of the recombinant type XVII collagen is 10 kDa to 90 kDa.

[0009] As a further embodiment of the present invention, the specific enzyme activity of the superoxide dismutase is 20,000 U / mg to 30,000 U / mg.

[0010] Technical theme two: The present invention provides a method for preparing a recombinant collagen filling powder for full-layer anti-aging, and the steps are as follows: S1: Add 40% to 60% of the volume of water used to glycerol, sodium hyaluronate and xanthan gum to obtain a mixed solution A; S2: Dissolve the recombinant collagen, superoxide dismutase, lysate of Saccharomyces cerevisiae and trehalose in the remaining water, and then add it to the mixed solution A for homogenization to obtain a mixed solution B; S3: Add silica, dimethylsilylated silica, polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer and dimethicone to the mixed solution B for homogenization to obtain a mixed solution C; S4: Add phenoxyethanol and ethylhexylglycerin to the mixed solution C for homogenization to obtain a white emulsion; S5: Take the white emulsion and perform spray drying at 30 to 45 °C, collect the dry powder obtained by spray drying, and obtain the recombinant collagen filling powder; The recombinant collagen is composed of recombinant type I collagen, recombinant type III collagen, recombinant type IV collagen, recombinant type VI collagen and recombinant type XVII collagen.

[0011] As a further embodiment of the present invention, the dosage ratio of the recombinant collagen to the superoxide dismutase is 8 g: 0.01 to 0.1 g.

[0012] As a further embodiment of the present invention, the mass ratio of the recombinant collagen to the lysate of Saccharomyces cerevisiae is 8: 1 to 20.

[0013] As a further embodiment of the present invention, the homogenization time is 3 to 8 min; the homogenization rate is 8000 to 12000 r / min.

[0014] Technical Theme Three: The present invention provides an application of the recombinant collagen filling powder with full-layer anti-aging as described in Technical Theme One in the field of skin care products.

[0015] The beneficial effects produced by adopting the above technical solutions are as follows: 1. The filling powder obtained in the present invention is added with various types of recombinant collagen, superoxide dismutase and lysate of saccharomyces boulardii. Through the combined action of recombinant collagen, superoxide dismutase and other components, a good anti-aging effect is achieved. When applied to skin care products, it can significantly increase the skin water content of users, improve skin firmness, and at the same time fade wrinkles.

[0016] 2. The recombinant collagen filling powder provided by the present invention constructs a collagen three-dimensional repair system by scientifically proportioning five types of collagen, namely recombinant type I collagen, recombinant type III collagen, recombinant type IV collagen, recombinant type VI collagen and recombinant type XVII collagen, and is added with superoxide dismutase to cooperate, giving full play to the anti-aging and repair effects of each type of collagen at different structural levels of the skin, and achieving the full-layer anti-aging effect. It can not only improve the epidermal aging of the skin by increasing the skin water content, but also improve the aging problem of the dermis layer of the skin to improve skin firmness. At the same time, it repairs the deep aging problems of the fat layer, fascia layer and muscle layer, and alleviates skin problems such as wrinkles and skin relaxation.

[0017] 3. The recombinant collagen filling powder obtained in the present invention is provided in the form of microcapsule powder. Microscopically, the filling powder presents a microcapsule structure, and the active ingredients are wrapped inside the microcapsule structure, which long-term maintains the biological activities of recombinant collagen, superoxide dismutase and saccharomyces boulardii, improves the stability of the obtained product, and extends its service life. At the same time, the microcapsule powder technology makes the recombinant collagen filling powder provided by the present invention easy to use. When in use, there is no need for cumbersome operations such as dissolving with a solvent. It can be directly applied to the skin surface. By pressing, the microcapsule structure is destroyed, and the active ingredients are released. The recombinant collagen filling powder turns into cream when smeared, fully releasing the active ingredients and exerting its efficacy. Description of the Drawings

[0018] Figure 1 It is a graph of the proliferation data of human immortalized keratinocytes in Test Example 1; Figure 2 It is a graph of the influence data of the filling powder obtained in Example 1 of the present invention on the ROS level of ultraviolet-damaged human immortalized keratinocytes; Figure 3 It is a graph of the anti-aging test results of the filling powder obtained in Example 1 of the present invention at the animal level; Figure 4 It is the microscopic structure diagram of the recombinant collagen filling powder obtained in Example 1 of the present invention. Detailed implementation mode

[0019] To make the objectives, technical solutions, and advantages of the present invention clearer, the following describes the invention clearly and completely in conjunction with specific embodiments. Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field to which the present invention belongs. The terms used in the description of the present invention herein are only for the purpose of describing specific implementation modes and are not intended to limit the present invention. For the experimental methods or test methods involved in the embodiments of the present invention, unless otherwise specified, they are all conventional methods in the prior art, and their names and / or abbreviations are all conventional names in the field, which are very clear and definite in the relevant application fields. Those skilled in the art can understand the conventional process steps according to the names and apply the corresponding equipment and implement them under conventional conditions or the conditions recommended by the manufacturer. There are no special restrictions on the sources of various instruments, equipment, raw materials, or reagents used in the embodiments of the present invention. They are all conventional products that can be obtained through regular commercial channels or can be prepared according to the conventional methods well-known to those skilled in the art.

[0020] Recombinant type I collagen is prepared by the method disclosed in Example 6 of the invention patent with the title of "A recombinant human type I collagen and its preparation method and application" and the patent number of 202411452076.8, and its molecular weight is about 35 kDa; Recombinant type III collagen is prepared by the method disclosed in Example 7 of the invention patent with the title of "A recombinant human type III collagen and its preparation method and uses" and the patent number of 202011366731.X, and its molecular weight is about 70 kDa; Recombinant type IV collagen is prepared by the method disclosed in Example 5 of the invention patent with the title of "A recombinant human type IV collagen and its preparation method and application" and the patent number of 202411316620.6, and its molecular weight is about 20 kDa; Recombinant type VI collagen is prepared according to the technical solution disclosed in Example 6 of the invention patent with the title of "A recombinant human type VI collagen and its coding gene and expression method" and the patent number of 2025104390743, and its molecular weight is about 30 kDa; Recombinant type XVII collagen is prepared by the method disclosed in Example 5 of the invention patent with the title of "A water-soluble recombinant type XVII collagen and its preparation method and application" and the patent number of 202410118032.5, and its molecular weight is about 17 kDa; Superoxide dismutase was prepared by the method disclosed in Example 9 of the invention patent with the Chinese patent number 202211658617.3, titled "A recombinant Cu, Zn-SOD with high enzyme activity and good stability, its preparation method and uses", and the specific activity of the enzyme was 20,000 U / mg.

[0021] Preparation Example 1: Recombinant human type VI collagen S1. Gene design and synthesis: According to the sequence of human type VI collagen, the amino acid sequence was designed (as shown in SEQ ID NO.1), the nucleic acid sequence was designed in reverse and codon-optimized, and Nde I and Kpn I restriction enzyme sites were added to obtain the nucleic acid sequence of recombinant human type VI collagen HC122 (as shown in SEQ ID NO.2), and the gene fragment was synthesized; SEQ ID NO.1: MHHHHHHGLDGEDGDKGLPGSSGEKGNPGRRGDKGPRGEKGERGDVGIRGDPGNPGQDSQERGPKGETGDLGPMGVPGRDGVPGGPGETGKNGGFGRRGPPGAKGNKGGPGQPGFEGEQGTRGAQGPAGPAGPPGLIGEQGISGPRGSGGAAGAPGERGRTGPLGRKGEPGEPGPKGGIGNRGPRGETGDDGRDGVGSEGRRGKKGERGFPGYPGPKGNPGEPGLNGTTGPKGIRGRRGNSGPPGIVGQKGDPGYPGPAGPKGNRGDSID.

[0022] SEQ ID NO.2: ATGGGGGTAAAAGGAGAAGCTGGTCTACCCGGAACCCCGGGTCCAACGGGTCCGGCAGGTCAGAAAGGCGAACCGGGTTCTGATGGTATCCCGGGCAGCGCAGGCGAAAAGGGCGAGCCGGGCCTCCCGGGACGCGGTTTCCCGGGTTTCCCGGGCGCGAAAGGCGACAAAGGCAGCAAGGGCGAGGTGGGCTTCGACGGTCGTAATGGTGAGAAGGGTGAAACCGGTGCGCCTGGTCTGAAAGGCGAGAATGGCTTGCCGGGCGAGAACGGCGCGCCGGGCCCGATGGGTCCGCGTGGTGCGCCAGGTGAGCGCGGTAGACCGGGCCTTCCGGGCGCTGCCGGCGCGCGTGGCGGGGCGGGCTCCCTGGGTGCCGGTGGCGCGTTTGGTGAAGCAGCGGGTGACCGTGGTCCGTACGGCACCGATATTGGTCCGGGCGGTGGTTATGGTGCCGCTGCTGAAGGTGGTATGTACGCGGGAAACGGCGGTTTGCTGGGTGCGGATTTTGCTGGCGACCTGGATTATAACGAACTGGCACATCATCACCACCACCATCACTAA。

[0023] S2: Construction of expression vector pET30a(+)-HC122: The nucleic acid fragment obtained in S1 was ligated to plasmid pET30a(+) through the multiple cloning sites NdeI and KpnI.

[0024] S3: Construction of expression strain BL21(DE3) / pET30a(+)-HC122: The expression vector obtained in S2 was transformed into competent Escherichia coli BL21(DE3) by heat shock method; The transformed cells were aspirated and spread on a plate containing 50 μg / mL antibiotic (kanamycin), and incubated upside down in a 37 °C incubator. The grown colonies were the expression strain BL21(DE3) / pET30a(+)-HC122.

[0025] S4: Induced expression of expression strain BL21(DE3) / pET30a(+)-HC122: IPTG (isopropyl-β-D-thiogalactoside) with a final concentration of 0.5 mM was added to the expression strain obtained in S3 for induced expression, and the cells were collected by centrifugation. S5: Purification of recombinant human type VI collagen HC122: Disrupt the bacterial cells and collect the supernatant by centrifugation; perform affinity chromatography using a Ni Sepharose FF column, and obtain recombinant human type VI collagen after desalting.

[0026] Example 1 A recombinant collagen filling powder for full-layer anti-aging, the preparation method is as follows: S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 236.2 g of water and stir to dissolve. The stirring time is 2 h, and the stirring rate is 1500 rpm to obtain a mixed solution A; S2: Weigh 6.4 g of recombinant type I collagen, 1.2 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, 10 g of lysate of Saccharomyces bispora, and 30 g of trehalose, add them to 236.2 g of water and stir to dissolve, then add them to the mixed solution A and perform high-speed homogenization. The homogenization time is 5 min, and the homogenization rate is 10000 rpm to obtain a mixed solution B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 30 g of dimethicone, add them to the mixed solution B and perform high-speed homogenization. The homogenization time is 5 min, and the homogenization rate is 10000 rpm to obtain a mixed solution C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to the mixed solution C and perform high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and perform low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filling powder; Perform microscopic observation on the recombinant collagen filling powder obtained in this example, and the results are as Figure 4 shown. The microstructure of the recombinant collagen filling powder obtained in this example presents a microcapsule-encapsulated form.

[0027] Example 2 A recombinant collagen filling powder for full-layer anti-aging, the preparation method is as follows: S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 236.2 g of water and stir to dissolve. The stirring time is 2 h, and the stirring rate is 1500 rpm to obtain a mixed solution A; S2: Weigh 6.8 g of recombinant type I collagen, 0.8 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, 10 g of bifidobacterium lysate and 30 g of trehalose, add them to 236.2 g of water and stir to dissolve. Then add them to the mixture A and carry out high-speed homogenization for 5 minutes at a homogenization rate of 10,000 rpm to obtain mixture B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer and 30 g of dimethicone, add them to the mixture B and carry out high-speed homogenization for 5 minutes at a homogenization rate of 10,000 rpm to obtain mixture C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to the mixture C and carry out high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and carry out low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filling powder.

[0028] Example 3 A preparation method of a recombinant collagen filling powder for full-layer anti-aging, comprising the following steps: S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate and 20 g of xanthan gum, add them to 236.2 g of water and stir to dissolve. The stirring time is 2 hours and the stirring rate is 1,500 rpm to obtain mixture A; S2: Weigh 6 g of recombinant type I collagen, 1.6 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, 10 g of bifidobacterium lysate and 30 g of trehalose, add them to 236.2 g of water and stir to dissolve. Then add them to the mixture A and carry out high-speed homogenization for 5 minutes at a homogenization rate of 10,000 rpm to obtain mixture B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer and 30 g of dimethicone, add them to the mixture B and carry out high-speed homogenization for 5 minutes at a homogenization rate of 10,000 rpm to obtain mixture C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to the mixture C and carry out high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and carry out low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filling powder.

[0029] Example 4 A preparation method of a recombinant collagen filling powder for full-layer anti-aging, comprising the following steps: S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 236.3 g of water, stir and dissolve for 2 h at a stirring rate of 1500 rpm to obtain a mixed solution A; S2: Weigh 6.4 g of recombinant type I collagen, 1.2 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 0.01 g of superoxide dismutase, 10 g of lysate of Saccharomyces boulardii, and 30 g of trehalose, add them to 236.3 g of water, stir and dissolve, then add them to the mixed solution A for high-speed homogenization for 5 min at a homogenization rate of 10000 rpm to obtain a mixed solution B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 30 g of dimethicone, add them to the mixed solution B for high-speed homogenization for 5 min at a homogenization rate of 10000 rpm to obtain a mixed solution C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerol, add them to the mixed solution C for high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion for low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filling powder.

[0030] Example 5 A preparation method of a recombinant collagen filling powder for full-layer anti-aging, comprising the following steps: S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 236.2 g of water, stir and dissolve for 2 h at a stirring rate of 1500 rpm to obtain a mixed solution A; S2: Weigh 6.4 g of recombinant type I collagen, 1.2 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 0.1 g of superoxide dismutase, 10 g of lysate of Saccharomyces boulardii, and 30 g of trehalose, add them to 236.2 g of water, stir and dissolve, then add them to the mixed solution A for high-speed homogenization for 5 min at a homogenization rate of 10000 rpm to obtain a mixed solution B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 30 g of dimethicone, add them to the mixed solution B for high-speed homogenization for 5 min at a homogenization rate of 10000 rpm to obtain a mixed solution C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to the mixed solution C, and perform high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and perform low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying, and obtain the recombinant collagen filler powder.

[0031] Example 6 A recombinant collagen filler powder for full-layer anti-aging, the preparation method is as follows: S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 241.0 g of water, stir and dissolve for 2 h at a stirring rate of 1500 rpm to obtain a mixed solution A; S2: Weigh 6.4 g of recombinant type I collagen, 1.2 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, 1 g of bifidobacterium lysate, and 30 g of trehalose, add them to 241.0 g of water, stir and dissolve, and then add them to the mixed solution A for high-speed homogenization for 5 min at a homogenization rate of 10000 rpm to obtain a mixed solution B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 30 g of dimethicone, add them to the mixed solution B for high-speed homogenization for 5 min at a homogenization rate of 10000 rpm to obtain a mixed solution C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to the mixed solution C, and perform high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and perform low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying, and obtain the recombinant collagen filler powder.

[0032] Example 7 A recombinant collagen filler powder for full-layer anti-aging, the preparation method is as follows: S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 231.0 g of water, stir and dissolve for 2 h at a stirring rate of 1500 rpm to obtain a mixed solution A; S2: Weigh 6.4 g of recombinant type I collagen, 1.2 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, 20 g of bifidobacterium lysate and 30 g of trehalose, add them to 231.0 g of water, stir to dissolve, then add them to the mixture A and perform high-speed homogenization for 5 minutes at a homogenization rate of 10,000 rpm to obtain mixture B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer and 30 g of dimethicone, add them to mixture B and perform high-speed homogenization for 5 minutes at a homogenization rate of 10,000 rpm to obtain mixture C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to mixture C and perform high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and perform low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filling powder.

[0033] Example 8 A recombinant collagen filling powder for full-layer anti-aging, the preparation method is as follows: S1: Weigh 80 g of glycerol, 0.2 g of sodium hyaluronate and 10 g of xanthan gum, add them to 343.9 g of water, stir to dissolve, and stir for 2 hours at a stirring rate of 1,500 rpm to obtain mixture A; S2: Weigh 6.4 g of recombinant type I collagen, 1.2 g of recombinant type III collagen, 0.008 g of recombinant type IV collagen, 0.008 g of recombinant type VI collagen, 0.20 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, 10 g of bifidobacterium lysate and 10 g of trehalose, add them to 343.9 g of water, stir to dissolve, then add them to mixture A and perform high-speed homogenization for 5 minutes at a homogenization rate of 10,000 rpm to obtain mixture B; S3: Weigh 100 g of silica, 50 g of dimethylsilylated silica, 30 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer and 10 g of dimethicone, add them to mixture B and perform high-speed homogenization for 5 minutes at a homogenization rate of 10,000 rpm to obtain mixture C; S4: Weigh 3 g of phenoxyethanol and 1 g of ethylhexylglycerin, add them to mixture C and perform high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and perform low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filling powder.

[0034] Example 9 A recombinant collagen filling powder for full-layer anti-aging, and the preparation method is as follows: S1: Weigh 200 g of glycerol, 0.8 g of sodium hyaluronate, and 30 g of xanthan gum, add them to 127.9 g of water and stir to dissolve. The stirring time is 2 h, and the stirring rate is 1500 rpm to obtain a mixed solution A; S2: Weigh 6.4 g of recombinant type I collagen, 1.2 g of recombinant type III collagen, 0.12 g of recombinant type IV collagen, 0.12 g of recombinant type VI collagen, 0.4 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, 10 g of lysate of saccharomyces boulardii, and 50 g of trehalose, add them to 127.9 g of water and stir to dissolve, then add them to the mixed solution A and perform high-speed homogenization. The homogenization time is 5 min, and the homogenization rate is 10000 rpm to obtain a mixed solution B; S3: Weigh 200 g of silica, 100 g of dimethylsilylated silica, 80 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 50 g of dimethicone, add them to the mixed solution B and perform high-speed homogenization. The homogenization time is 5 min, and the homogenization rate is 10000 rpm to obtain a mixed solution C; S4: Weigh 5 g of phenoxyethanol and 10 g of ethylhexylglycerin, add them to the mixed solution C and perform high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and perform low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filling powder.

[0035] Comparative Example 1 S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 236.2 g of water and stir to dissolve. The stirring time is 2 h, and the stirring rate is 1500 rpm to obtain a mixed solution A; S2: Weigh 7.2 g of recombinant type I collagen, 0.4 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, 10 g of lysate of saccharomyces boulardii, and 30 g of trehalose, add them to 236.2 g of water and stir to dissolve, then add them to the mixed solution A and perform high-speed homogenization. The homogenization time is 5 min, and the homogenization rate is 10000 rpm to obtain a mixed solution B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 30 g of dimethicone, add them to the mixed solution B and perform high-speed homogenization. The homogenization time is 5 min, and the homogenization rate is 10000 rpm to obtain a mixed solution C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to the mixed solution C, and perform high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and perform low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying, and obtain the recombinant collagen filler powder.

[0036] Comparative Example 2 S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 236.2 g of water, stir and dissolve for 2 h at a stirring rate of 1500 rpm to obtain the mixed solution A; S2: Weigh 5.6 g of recombinant type I collagen, 2 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, 10 g of lysate of Saccharomyces cerevisiae, and 30 g of trehalose, add them to 236.2 g of water, stir and dissolve, then add them to the mixed solution A and perform high-speed homogenization for 5 min at a homogenization rate of 10000 rpm to obtain the mixed solution B; S2: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 30 g of dimethicone, add them to the mixed solution B and perform high-speed homogenization for 5 min at a homogenization rate of 10000 rpm to obtain the mixed solution C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to the mixed solution C, and perform high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and perform low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying, and obtain the recombinant collagen filler powder.

[0037] Comparative Example 3 S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 236.3 g of water, stir and dissolve for 2 h at a stirring rate of 1500 rpm to obtain the mixed solution A; S2: Weigh 6.4 g of recombinant type I collagen, 1.2 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.24 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, 10 g of lysate of Saccharomyces cerevisiae, and 30 g of trehalose, add them to 236.3 g of water, stir and dissolve, then add them to the mixed solution A and perform high-speed homogenization for 5 min at a homogenization rate of 10000 rpm to obtain the mixed solution B; S2: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 30 g of dimethicone, add them to mixture B, and conduct high-speed homogenization for 5 min at a homogenization rate of 10,000 rpm to obtain mixture C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to mixture C, and conduct high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and conduct low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filler powder.

[0038] Comparative Example 4 S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 244.2 g of water, stir and dissolve for 2 h at a stirring rate of 1,500 rpm to obtain mixture A; S2: Weigh 0.05 g of superoxide dismutase, 10 g of lysate of Saccharomyces boulardii, and 30 g of trehalose, add them to 244.2 g of water, stir and dissolve, and then add them to mixture A and conduct high-speed homogenization for 5 min at a homogenization rate of 10,000 rpm to obtain mixture B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 30 g of dimethicone, add them to mixture B, and conduct high-speed homogenization for 5 min at a homogenization rate of 10,000 rpm to obtain mixture C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to mixture C, and conduct high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and conduct low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the filler powder.

[0039] Comparative Example 5 S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 236.3 g of water, stir and dissolve for 2 h at a stirring rate of 1,500 rpm to obtain mixture A; S2: Weigh 6.4 g of recombinant type I collagen, 1.2 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 10 g of lysate of Saccharomyces boulardii, and 30 g of trehalose, add them to 236.3 g of water, stir and dissolve, and then add them to mixture A and conduct high-speed homogenization for 5 min at a homogenization rate of 10,000 rpm to obtain mixture B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 30 g of dimethicone, add them to mixture B, and perform high-speed homogenization for 5 min at a homogenization rate of 10,000 rpm to obtain mixture C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to mixture C, and perform high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and perform low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filler powder.

[0040] Comparative Example 6 S1: Weigh 140 g of glycerol, 0.5 g of sodium hyaluronate, and 20 g of xanthan gum, add them to 246.2 g of water, stir and dissolve for 2 h at a stirring rate of 1500 rpm to obtain mixture A; S2: Weigh 6.4 g of recombinant type I collagen, 1.2 g of recombinant type III collagen, 0.08 g of recombinant type IV collagen, 0.08 g of recombinant type VI collagen, 0.24 g of recombinant type XVII collagen, 0.05 g of superoxide dismutase, and 30 g of trehalose, add them to 246.2 g of water, stir and dissolve, then add them to mixture A and perform high-speed homogenization for 5 min at a homogenization rate of 10,000 rpm to obtain mixture B; S3: Weigh 150 g of silica, 80 g of dimethylsilylated silica, 50 g of polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer, and 30 g of dimethicone, add them to mixture B, and perform high-speed homogenization for 5 min at a homogenization rate of 10,000 rpm to obtain mixture C; S4: Weigh 4 g of phenoxyethanol and 5 g of ethylhexylglycerin, add them to mixture C, and perform high-speed homogenization to obtain a white emulsion; S5: Take the white emulsion and perform low-temperature spray drying at 40 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filler powder.

[0041] Investigation Example 1: Effect of different types and ratios of recombinant collagen on the proliferation of HaCaT cells (human immortalized keratinocytes) Seed HaCaT cells into a 96-well plate at a density of 3×10 3 cells per well. After culturing the seeded cells in a cell culture incubator at 37 °C and 5% CO2 for 24 h, dissolve the sample with cell culture medium at a sample concentration of 0.1 mg / ml to obtain a sample solution; add 100 μL of the sample solution to each well and continue culturing in the incubator for 24 h; add 10 μL of CCK8 reagent to each well and continue culturing in the incubator for 2 h. Use an enzyme-linked immunosorbent assay (ELISA) reader to detect the absorbance value of each group of HaCaT cells at OD = 450 nm.

[0042] Among them, Control group: no sample was added; Sample of Group 1: recombinant type I collagen; Sample of Group 2: recombinant type III collagen; Sample of Group 3: superoxide dismutase (SOD); Sample of Group 4: a mixture of recombinant type IV collagen, recombinant type VI collagen, recombinant type XVII collagen and SOD with a mass ratio of 1:1:3:0.6; Sample of Group 5: a mixture of recombinant type I collagen, recombinant type III collagen, recombinant type IV collagen, recombinant type VI collagen, recombinant type XVII collagen and SOD with a mass ratio of 80:15:1:1:3:0.6; Sample of Group 6: a mixture of recombinant type I collagen, recombinant type III collagen, recombinant type IV collagen, recombinant type VI collagen, recombinant type XVII collagen and SOD with a mass ratio of 85:10:1:1:3:0.6; Sample of Group 7: a mixture of recombinant type I collagen, recombinant type III collagen, recombinant type IV collagen, recombinant type VI collagen, recombinant type XVII collagen and SOD with a mass ratio of 75:20:1:1:3:0.6; Sample of Group 8: a mixture of recombinant type I collagen, recombinant type III collagen, recombinant type IV collagen, recombinant type VI collagen, recombinant type XVII collagen and SOD with a mass ratio of 90:5:1:1:3:0.6; Sample of Group 9: a mixture of recombinant type I collagen, recombinant type III collagen, recombinant type IV collagen, recombinant type VI collagen, recombinant type XVII collagen and SOD with a mass ratio of 70:25:1:1:3:0.6.

[0043] The results are as Figure 1 shown that after the recombinant collagen and SOD were mixed, the performance of promoting the proliferation of HaCaT cells was significantly improved compared with the use alone; among them, the sample of Group 5: a mixture of recombinant type I collagen, recombinant type III collagen, recombinant type IV collagen, recombinant type VI collagen, recombinant type XVII collagen and SOD with a mass ratio of 80:15:1:1:3:0.6 had a more obvious ability to promote the proliferation of HaCaT cells compared with other groups of recombinant collagen and SOD mixtures, and had a significant ability to promote the proliferation of HaCaT cells.

[0044] Effect Example 1: Evaluation of anti-aging effect at the cellular level HaCaT cells were inoculated in a 96-well plate at a density of 3×10 3 cells per well.

[0045] The inoculated HaCaT cells were divided into a blank group, a control group, an experimental group, and a VC group, and the treatments for each group were as follows: Control group: Cultured in a cell incubator at 37°C and 5% CO2 for 12 h, and the HaCaT cells were not irradiated with UVB. Blank group: Cultured in a cell incubator at 37°C and 5% CO2 for 12 h, and the HaCaT cells were irradiated with 40 mJ / cm 2 UVB for 30 min without sample treatment. Experimental group: The recombinant collagen filler powder obtained in Example 1 of the present invention was dissolved in the complete cell culture medium of DMEM for HaCaT cells, and the final concentration was 0.1 mg / ml. After adding the recombinant collagen filler powder and culturing HaCaT cells in a cell incubator at 37°C and 5% CO2 for 12 h, 40 mJ / cm 2 UVB was used for irradiation for 30 min. VC group: VC powder was dissolved in the complete cell culture medium of DMEM for HaCaT cells at a concentration of 0.1 mg / ml; after adding VC powder, HaCaT cells were cultured in a cell incubator at 37°C and 5% CO2 for 12 h, and 40 mJ / cm 2 UVB was used for irradiation for 30 min.

[0046] After the HaCaT cells were treated, they were placed in a cell incubator and cultured for 24 h, then washed 3 times with PBS. 100 μl of serum-free medium containing 10 μmol / L DCFH-DA was added to each well, and incubated at 37°C in the dark for 30 min. The absorbance value of the HaCaT cells was detected by an enzyme-labeled instrument. Highly active reactive oxygen species (ROS) in the body can induce mitochondrial DNA damage and cell apoptosis, and lead to abnormal regulation of tissue differentiation, proliferation, and damage inflammation processes, promoting collagen degradation, thus resulting in skin photoaging.

[0047] The ROS level in the cells was determined according to the absorbance value, and the formula was as follows.

[0048] The results were as Figure 2 shown. Compared with the blank group, the control group significantly increased the ROS level in the cells ( *** P <0.001). Compared with the control group, the experimental group and the VC group significantly reduced the ROS level in the cells ( ## P <0.01). Importantly, the ability of the experimental group and the VC group with the same concentration to scavenge cellular ROS was comparable.

[0049] Effect Example 2: Evaluation of anti-aging effect at the animal level β-galactosidase is a hydrolase found in lysosomes. Its activity is higher in aging cells and it is one of the most commonly used markers of senescent cells. Zebrafish share similarities with humans in gene expression and protein function. Therefore, zebrafish can be used for anti-aging efficacy experiments, and the data obtained can provide a reliable basis for evaluating the anti-aging efficacy of cosmetics.

[0050] In this experiment, AB zebrafish embryos 8 hours after fertilization were randomly divided into a blank control group, experimental group 1, experimental group 2, and experimental group 3, and placed in different groups of culture tanks: Among them, the blank control group did not add any sample; Experimental Group 1: Add the recombinant collagen filling powder sample obtained in Example 1 to the culture tank, with a final concentration of 5 μg·mL −1 ; Experimental Group 2: Add the recombinant collagen filling powder sample obtained in Example 1 to the culture tank, with a final concentration of 25 μg·mL −1 ; Experimental Group 3: Add the recombinant collagen filling powder sample obtained in Example 1 to the culture tank, with a final concentration of 50 μg·mL −1 .

[0051] After placing the culture tanks of each group in a constant temperature incubator at 28.5°C for 72 hours, the β-galactosidase in the zebrafish was stained using a β-galactosidase staining kit, and the cells were observed under a microscope to measure the staining intensity.

[0052] The results are as follows Figure 3 As shown, compared with the control group, the activity of β-galactosidase in zebrafish of experimental groups 1-3 was significantly reduced ( P <0.01); Experimental results show that the recombinant collagen filling powder obtained by the present invention has anti-aging effect at the animal level.

[0053] Effect Example 3: Evaluation of Anti-aging Effects on Human Body 1. Skin moisture content change test: The decrease in skin moisture content is a direct manifestation of aging in the skin's epidermis.

[0054] Female volunteers aged 40-55 years old, with healthy skin and no history of skin allergies who met the inclusion criteria were selected, with 10 subjects in each group. Testing method: A square area of 4 cm in length and width was marked on the left and right cheeks of the subjects, and the test was performed at 3 mg / cm 2 The filling powder obtained in Examples 1 to 9 or Comparative Examples 1 to 6 was evenly applied on a square area of the subject's cheek; a skin moisture meter was used to detect changes in skin moisture content before application, 1 hour after application, and 4 hours after application.

[0055] Table 1 Evaluation of Skin Moisturizing Effect after Use From the data in Table 1, it can be seen that the skin moisture of the subjects after applying the recombinant collagen filling powder obtained in Example 1 of the present invention for 1 h and 4 h has a higher growth rate compared to the skin moisture of the subjects applying the filling powder obtained in the comparative example; among them, the skin moisture of the subjects applying the recombinant collagen filling powder obtained in Example 1 has the most obvious increase, indicating that the recombinant collagen filling powder obtained in the present invention has a good skin moisturizing effect.

[0056] 2. Skin Elasticity and Firmness Change Test The aging of the skin dermis is manifested by a decrease in skin elasticity and a change in firmness.

[0057] Female volunteer subjects aged 40 - 55 years old, with healthy skin, no skin allergy history and meeting the subject inclusion criteria were selected, with 10 subjects in each group.

[0058] After cleansing the face, an appropriate amount of the filling powder obtained in Examples 1 - 9 or Comparative Examples 1 - 6 was taken and evenly applied to the face, evenly applied and massaged until absorbed, once in the morning and once in the evening every day. On the 0th day and the 28th day of using the essence powder, the effects of skin elasticity, firmness and wrinkle degree were evaluated.

[0059] The skin elasticity (evaluated by the R2 value, the larger the R2 value, the better the skin elasticity) and skin firmness (evaluated by the F4 value, the smaller the F4 value, the better the skin firmness) of the cheek area of the subjects on the 0th day and the 28th day of use were measured by the skin elasticity tester Cutometer MPA580. The skin R2 values and skin F4 values of each group of subjects are shown in Table 2.

[0060] The values of each group are expressed using the change rate, and the formula is as follows.

[0061] Table 2 Evaluation of Skin Elasticity and Firmness before and after Use From the data in Table 2, it can be seen that after 28 days of using the filling powder obtained in Examples 1 - 9 or Comparative Examples 1 - 6, the improvement effect of the filling powder group of the examples on skin firmness and elasticity is more obvious than that of the filling powder group of the comparative examples, and the skin elasticity and firmness are significantly improved; among them, the effect of Example 1 is the most obvious, indicating that the recombinant collagen filling powder obtained in the present invention can significantly improve and restore skin elasticity, tighten the skin, slow down aging, and has a good anti - aging and firming effect.

[0062] 3. Skin Roughness and Texture Change Test Deep aging of the skin's adipose layer, fascia layer, and muscle layer leads to the formation of wrinkles, nasolabial folds, tear troughs, eye bags, and sunken apple muscles, etc., which in turn affect skin texture and roughness.

[0063] Female volunteer subjects aged 40 - 55 years old, with healthy skin, no history of skin allergies, and meeting the subject inclusion criteria were selected, with 10 subjects in each group. The skin roughness (characterized by the Sa value, the smaller the Sa value, the lower the skin roughness and the fewer skin wrinkles) and texture non-uniformity (characterized by the Sq value, the smaller the Sq value, the smaller the skin texture non-uniformity and the fewer skin wrinkles) of the cheek area of the subjects were measured by the skin tester Visioscan® VC98 at 0d and 28d after using the filling powders obtained in Examples 1 - 9 or Comparative Examples 1 - 6. The change rates of the Sa value and Sq value of the skin anti-wrinkle effect of each group of subjects are shown in Table 3.

[0064] The values of each group are represented by the change rate, and the formula is as follows.

[0065] Table 3 Evaluation of skin anti-wrinkle effect before and after use From the data in Table 3, it can be seen that 28 days after the subjects used the filling powders obtained in Examples 1 - 9 or Comparative Examples 1 - 6, the filling powder group of the examples had a more obvious effect on reducing the skin roughness and texture non-uniformity than the filling powder group of the comparative examples. The skin wrinkles were significantly reduced and smoothed, showing a good skin anti-wrinkle effect. Among them, the effect of Example 1 was the most obvious, indicating that the recombinant collagen filling powder obtained in the present invention can significantly improve skin roughness and reduce skin wrinkles, and has a good anti-aging and anti-wrinkle effect.

[0066] This effect example proves through testing that the recombinant collagen filling powder obtained in the present invention can significantly increase the skin water content of the subjects and has an obvious repair effect on epidermal skin aging; this effect example proves through testing that the recombinant collagen filling powder obtained in the present invention can significantly improve the skin elasticity and firmness of the subjects and has an improvement effect on the middle and even deep skin aging.

[0067] In summary, the recombinant collagen filling powder obtained in the present invention adopts the microcapsule-encapsulated powder cream technology to fully maintain the biological activity of the product and is convenient to use; the multi-type recombinant collagen is scientifically proportioned and used in combination with various components such as superoxide dismutase and lysate of Saccharomyces cerevisiae. The components synergistically enhance the effect, giving full play to the full-layer anti-aging effect, optimizing and repairing layer by layer, and having a good anti-aging and anti-wrinkle effect. Applying it to the fields of skin care products or medical aesthetics, etc., can generate good economic benefits and market value.

[0068] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A recombinant collagen filling powder for full-layer anti-aging, characterized in that, Containing the following components: Recombinant type I collagen with a mass fraction of 0.6% - 0.7%; Recombinant type III collagen with a mass fraction of 0.08% - 0.16%; Recombinant type IV collagen with a mass fraction of 0.008% - 0.012%; Recombinant type VI collagen with a mass fraction of 0.008% - 0.012%; Recombinant type XVII collagen with a mass fraction of 0.02% - 0.04%; Superoxide dismutase with a mass fraction of 0.001% - 0.01%; Lyophilized bifidobacterium lysate with a mass fraction of 0.1% - 2%; Trehalose with a mass fraction of 1% - 5%; Silica with a mass fraction of 10% - 20%; Dimethylsilylated silica with a mass fraction of 5% - 10%; Polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer with a mass fraction of 3% - 8%; Glycerol with a mass fraction of 8% - 20%; Xanthan gum with a mass fraction of 1% - 3%; Sodium hyaluronate with a mass fraction of 0.02% - 0.08%; Dimethicone with a mass fraction of 1% - 5%; Phenoxyethanol with a mass fraction of 0.3% - 0.5%; Ethylhexylglycerin with a mass fraction of 0.1% - 1%; And the balance is water.

2. The recombinant collagen filling powder for full-layer anti-aging according to claim 1, wherein The molecular weight of the recombinant type I collagen is 10 kDa - 90 kDa; the molecular weight of the recombinant type III collagen is 10 kDa - 90 kDa; the molecular weight of the recombinant type IV collagen is 10 kDa - 90 kDa; the molecular weight of the recombinant type VI collagen is 10 kDa - 90 kDa; the molecular weight of the recombinant type XVII collagen is 10 kDa - 90 kDa.

3. The recombinant collagen filling powder for full-layer anti-aging according to claim 1, wherein The enzyme specific activity of the superoxide dismutase is 20000 U / mg - 30000 U / mg.

4. A preparation method of a recombinant collagen filling powder for full-layer anti-aging, characterized in that, The steps are as follows: S1: Add 40% - 60% of the total water volume to glycerol, sodium hyaluronate and xanthan gum to obtain mixture A; S2: Dissolve the recombinant collagen, superoxide dismutase, lyophilized bifidobacterium lysate and trehalose in the remaining water, then add to mixture A and homogenize to obtain mixture B; S3: Add silica, dimethylsilylated silica, polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymer and dimethicone to mixture B and homogenize to obtain mixture C; S4: Add phenoxyethanol and ethylhexylglycerin to mixture C and homogenize to obtain a white emulsion; S5: Take the white emulsion and perform spray drying at 30 - 45 °C, collect the dry powder obtained by spray drying to obtain the recombinant collagen filler powder; The recombinant collagen is composed of recombinant type I collagen, recombinant type III collagen, recombinant type IV collagen, recombinant type VI collagen and recombinant type XVII collagen.

5. The preparation method according to claim 4, wherein The dosage ratio of the recombinant collagen to the superoxide dismutase is 8 g: 0.01 - 0.1 g.

6. The preparation method according to claim 4, characterized in that, The mass ratio of the recombinant collagen to the lyophilized bifidobacterium lysate is 8: 1 - 20.

7. The preparation method according to claim 4, wherein The homogenization time is 3 - 8 min; the homogenization rate is 8000 - 12000 r / min.

8. Use of the recombinant collagen filling powder with full-layer anti-aging as described in any one of claims 1 to 3 in the field of skin care products.

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