Cosmetic containing recombinant collagen composition and preparation method thereof

Abstract

The invention discloses a cosmetic containing a recombinant collagen composition and a preparation method of the cosmetic. The cosmetic can be a cosmetic containing an emulsifying system, such as a moisturizer, a body lotion, a face cream, an eye cream or a smearing mask. The cosmetic contains collagen functional components, and the recombinant III-type collagen and APG tripeptide have a synergistic effect in the aspects of keeping skin elasticity and resisting wrinkles and tightening; the recombinant I-type collagen and the LPG tripeptide have a synergistic effect in the aspects of increasing the thickness of a dermis layer of skin and promoting skin repair.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a cream cosmetic containing a recombinant collagen composition and its preparation method. Background Technology

[0002] Collagen is the most abundant protein in mammals, accounting for one-third of the human body's protein content and three-quarters of the dry weight of skin. It is also a major protein in the human extracellular matrix, widely present in skin, bones, joints, and hair, playing a vital role in support, repair, and anti-aging, while also providing essential structural and mechanical support for mammalian tissues. Due to its unique functions and properties, it is widely used in biomedicine, cosmetics, and biotechnology. Recombinant collagen molecules are simple, structurally clear, and easily controlled, making it the best alternative to animal-derived collagen in biomedicine and tissue engineering.

[0003] In the cosmetics industry, collagen is known as "soft gold for the skin." It exists in the dermis, making up about 70% of skin tissue. Small-molecule collagen can moisturize the skin, replenish essential amino acids, and repair damaged skin. Adding collagen to cosmetics provides essential nutrients to the skin, and its abundant hydrophilic groups give it excellent moisturizing properties. Additionally, collagen can stabilize foam, regulate and stabilize pH, emulsify colloids, and reduce the irritation of surfactants, acids, and alkalis on the skin and hair. If the collagen content in the deep and superficial layers of the skin is not replenished, the reduction in collagen content in the skin tissue will cause the skin to sag, lose elasticity, and become rough and dull.

[0004] Face cream is a common skincare product in daily skincare routines. It can be applied directly to the skin, greatly increasing the absorption of active ingredients and improving the longevity of moisturizing effects. However, current face cream cosmetics generally suffer from a lack of functional synergy between different types of collagen and an imbalance in collagen replenishment across different layers of skin tissue. Summary of the Invention

[0005] In view of the technical problems existing in the prior art, the object of the present invention is to provide a recombinant collagen-peptide composition with synergistic firming and anti-aging effects, and a cosmetic product containing an emulsification system that uses the recombinant collagen-peptide composition as a raw material and can evenly nourish the skin at all levels, such as a cream-type skin care product.

[0006] The inventors conducted in-depth research on the synergistic effects of various ingredients with firming and anti-aging properties, and found that: 1. A certain proportion of the recombinant type III collagen and APG tripeptide described in Chinese patent application 201310701942.8 have a synergistic effect in increasing fibroblast growth factor (b-FGF), thereby better maintaining and restoring skin elasticity and achieving anti-wrinkle and firming effects. 2. It was discovered for the first time that LPG tripeptide has the function of increasing the thickness of the dermal layer of the skin and promoting skin repair, and a certain proportion of the recombinant type I collagen and LPG tripeptide described in Chinese patent application 201310701767.2 have a synergistic effect in increasing the thickness of the dermal layer of the skin and promoting skin repair. Based on the above scientific discoveries, the inventors created the recombinant collagen-small peptide composition of the present invention, and further combined it with other functional ingredients of cosmetics to prepare the cosmetic containing an emulsification system of the present invention.

[0007] That is, the present invention includes:

[0008] 1. A cosmetic product comprising a recombinant collagen-peptide composition, and one or more selected from recombinant human collagen, skin conditioning agents, emulsifiers, antioxidants, emollients, moisturizers, thickeners, complexing agents, and solvents; wherein,

[0009] The recombinant collagen-peptide composition consists of recombinant type III collagen, recombinant type I collagen, APG tripeptide, and LPG tripeptide.

[0010] In the recombinant collagen-peptide composition, the weight ratio of recombinant type III collagen to APG tripeptide is 250:1 to 5:1, and / or the weight ratio of recombinant type I collagen to LPG tripeptide is 250:1 to 5:1.

[0011] The amino acid sequence of the recombinant type III collagen is shown in SEQ ID No:1;

[0012] The amino acid sequence of the recombinant type I collagen is shown in SEQ ID No:2;

[0013] The amino acid sequence of the recombinant human collagen is shown in SEQ ID No:3.

[0014] In this invention, both the recombinant collagen-peptide composition and the recombinant human collagen can be used as collagen-based functional ingredients in cosmetics, that is, the collagen-based functional ingredients can include the recombinant collagen-peptide composition and the recombinant human collagen.

[0015] 2. The cosmetic product according to item 1, wherein the weight ratio of the recombinant type III collagen to the APG tripeptide is 25:1 to 5:1.

[0016] 3. The cosmetic product according to item 1, wherein the weight ratio of the recombinant type I collagen to the LPG tripeptide is 25:1 to 5:1.

[0017] 4. The cosmetic according to claim 1, wherein the skin conditioning agent is one or more combinations selected from acetyl hexapeptide-8, 1,3-propanediol, chlorella extract, and mulberry extract.

[0018] 5. The cosmetic according to claim 1, wherein the emulsifier is one or more selected from cetearyl alcohol, behenyl alcohol, tribenzyl alcohol and arachidyl alcohol.

[0019] 6. The cosmetic according to claim 1, wherein the antioxidant is selected from p-hydroxyacetophenone and tocopheryl acetate.

[0020] 7. The cosmetic according to claim 1, wherein the emollient is one or more selected from the group consisting of caprylic / capric triglyceride, isononyl isononanoate, polydimethylsiloxane, squalane, ethyl ethyl glycerin, and tocopheryl acetate.

[0021] 8. The cosmetic product according to claim 1, wherein the moisturizer is one or more selected from glycerin, soluble collagen, 1,2-pentanediol, 1,2-hexanediol, caprylyl glycol, and sodium hyaluronate.

[0022] 9. The cosmetic according to claim 1, wherein the thickener is one or more selected from hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, acrylate / C10-30 alkanol acrylate crosspolymer.

[0023] 10. The cosmetic according to claim 1, wherein the complexing agent is selected from disodium EDTA and phytic acid.

[0024] 11. The cosmetic product according to item 1, which is a moisturizing cream, lotion, face cream, eye cream or face mask.

[0025] 12. The cosmetic product according to claim 1, which is a face cream, comprises:

[0026] 1-20 parts by weight of the recombinant collagen-peptide composition;

[0027] 1-20 parts by weight of recombinant human collagen;

[0028] 1-10 parts by weight of glycerin;

[0029] 1,2-Pentanediol 1-3 parts by weight;

[0030] 1-10 parts by weight of isononyl isononanoate;

[0031] Cetearyl alcohol 1-5 parts by weight;

[0032] Behenyl alcohol 1-5 parts by weight;

[0033] 1-10 parts by weight of squalane;

[0034] Tocopheryl acetate 0.5-5 parts by weight;

[0035] 0.1-3 parts by weight of 1,2-hexanediol;

[0036] Dipotassium glycyrrhizate 0.1-1 parts by weight;

[0037] Carnosine 0.1-2 parts by weight;

[0038] 1-5 parts by weight of *Pseudomonas leucantha* extract;

[0039] 0.1-3 parts by weight of acrylate / C10-30 alkanol acrylate crosspolymer;

[0040] Sodium hyaluronate 0.1-3 parts by weight;

[0041] 1-10 parts by weight of butanediol;

[0042] Disodium EDTA 0.01-1 parts by weight;

[0043] Acetyl hexapeptide-8 0.1-10 parts by weight;

[0044] Tocopheryl acetate 0.01-5 parts by weight;

[0045] And, an appropriate amount of purified water.

[0046] 13. The cosmetic product according to item 12, wherein the sodium hyaluronate has a molecular weight of 0.5-1.2 million Daltons.

[0047] 14. The cosmetic product according to claim 1, which is a topical mask, comprises:

[0048] 1-20 parts by weight of the recombinant collagen-peptide composition;

[0049] 1-20 parts by weight of recombinant human collagen;

[0050] 1-10 parts by weight of glycerin;

[0051] 1,2-Pentanediol 1-3 parts by weight;

[0052] 1-10 parts by weight of isononyl isononanoate;

[0053] Cetearyl alcohol 1-5 parts by weight;

[0054] 1-10 parts by weight of butanediol;

[0055] Sodium hyaluronate 0.1-2 parts by weight;

[0056] Behenyl alcohol 1-5 parts by weight;

[0057] 1-10 parts by weight of squalane;

[0058] Tocopheryl acetate 0.5-5 parts by weight;

[0059] 0.1-2 parts by weight of hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer;

[0060] Mulberry extract, 0.1-10 parts by weight;

[0061] And, an appropriate amount of purified water.

[0062] 15. The cosmetic product according to claim 1, which is an eye cream, comprises:

[0063] 1-20 parts by weight of the recombinant collagen-peptide composition;

[0064] 1-20 parts by weight of recombinant human collagen;

[0065] 1-10 parts by weight of glycerin;

[0066] 1,2-Pentanediol 1-3 parts by weight;

[0067] 1-10 parts by weight of isononyl isononanoate;

[0068] Cetearyl alcohol 1-5 parts by weight;

[0069] Behenyl alcohol 1-5 parts by weight;

[0070] 1-10 parts by weight of squalane;

[0071] Tocopheryl acetate 0.1-5 parts by weight;

[0072] 0.1-5 parts by weight of 1,2-hexanediol;

[0073] Dipotassium glycyrrhizate 0.1-2 parts by weight;

[0074] Carnosine 0.1-2 parts by weight;

[0075] 1-10 parts by weight of *Pseudomonas leucantha* extract;

[0076] 1-5 parts by weight of acrylate / C10-30 alkanol acrylate crosspolymer;

[0077] Sodium hyaluronate 0.1-3 parts by weight;

[0078] 1-10 parts by weight of butanediol;

[0079] Disodium EDTA 0.01-1 parts by weight;

[0080] Chlorella extract, 1-10 parts by weight;

[0081] Mulberry extract, 0.1-5 parts by weight;

[0082] Acetyl hexapeptide-8 1-20 parts by weight;

[0083] Tocopheryl acetate 0.01-5 parts by weight; and

[0084] An appropriate amount of purified water.

[0085] The cosmetic of the present invention can be prepared using conventional cosmetic preparation methods that include emulsification systems. For example...

[0086] 16. The method for preparing the face cream according to item 12, comprising the following steps:

[0087] (1) Disperse the extract of *Ilex pubescens*, butylene glycol, glycerin, 1,2-pentanediol, recombinant collagen-peptide composition, recombinant human collagen, acrylate / C10-30 alkanol acrylate crosspolymer, sodium hyaluronate, and disodium EDTA evenly, mix with purified water, add to the main pot, stir evenly, heat, and keep warm at 85±2℃ for 30 min;

[0088] (2) Dissolve squalane, isononyl isononanoate, cetearyl alcohol, behenyl alcohol, and tocopheryl acetate by heating, and keep warm at 80-85℃ for 10 min;

[0089] (3) After the heat preservation is completed, add the raw materials mentioned in (2) into the main pot, stir and homogenize until the material is uniform, add the pre-dispersed isononyl isononanoate and hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer into the main pot, and continue to emulsify and stir until the material is uniform.

[0090] (4) Cool down to 45℃-50℃, add the dissolved carnosine and other raw materials to the main pot, and stir evenly. Add acetyl hexapeptide-8 to the main pot, continue stirring and cooling down to below 30℃ to obtain the final product.

[0091] Compared with the prior art, the beneficial effects of the present invention include:

[0092] (1) In the recombinant collagen-small peptide composition, recombinant type III collagen and APG tripeptide have a synergistic effect in maintaining skin elasticity and anti-wrinkle firming, and recombinant type I collagen and LPG tripeptide have a synergistic effect in increasing the thickness of the dermis and promoting skin repair. This makes the recombinant collagen-small peptide composition and the cosmetic of the present invention have better firming and anti-aging effects.

[0093] (2) In addition to providing tissue flexibility and playing a protective role against tearing, type I collagen can also promote the growth of epithelial cells and play a key role in skin repair; type III collagen can promote the growth of fibroblasts and promote the secretion of collagen by fibroblasts, thereby maintaining and repairing skin elasticity and achieving anti-wrinkle and firming effects; sodium hyaluronate is a naturally occurring polysaccharide in the human body with excellent moisturizing and water-retaining functions. Sodium hyaluronate can absorb and lock in the moisture on the skin surface, forming a moisturizing film, thereby maintaining the skin's moisture and hydration. At the same time, it can prevent harmful substances and bacteria from invading the skin, playing a protective role, improving the skin's self-repair ability, and preventing and alleviating the occurrence of skin problems. The combined use of collagen and sodium hyaluronate can play a dual role, providing a long-lasting moisturizing effect while enhancing the role of collagen; collagen can more effectively play its role in improving skin elasticity and firmness, while sodium hyaluronate, with the support of collagen, can provide the skin with the required moisture and hydration for a long time, making the skin more supple and plump.

[0094] (3) The cosmetic of the present invention contains type I / type III collagen, which are very similar to the protein components that make up the deep skin. They can be quickly absorbed by the skin, replenish collagen, fade melanin, reduce wrinkles, and help skin cells synthesize and repair damaged collagen, elastic fibers and polysaccharides, thereby strengthening their structure, vitality and cell density, and enhancing their ability to lock in water and help synthesize collagen.

[0095] (3) The cosmetic of this invention contains sodium hyaluronate, a highly moisturizing, oxygenating, and brightening ingredient. Sodium hyaluronate is composed of sodium hyaluronate with a molecular weight of 0.5-1.2 million Daltons, which can inject energy into the superficial collagen tissue of the skin, and in combination with protecting the skin surface, improving skin aging, and strengthening skin connections to make it plump and hydrated. Finally, a recombinant collagen-peptide composition is used to increase the collagen and moisture content of the skin's basal layer, improve rough skin and enlarged pores, while firming, anti-wrinkle, and slowing down skin aging.

[0096] (4) The cosmetic of the present invention can simultaneously replenish type I / III collagen, achieving gradual brightening, intensive moisturizing, firming and reducing wrinkles, so that the skin presents a state of full collagen. Attached Figure Description

[0097] Figure 1 A figure showing the effect of face cream on reactive oxygen species (ROS) in zebrafish embryos (p = 0.000060). Figure 2 Figure showing the effect of face cream on the expression of collagen-related genes in zebrafish embryos (**** indicates p < 0.001). Detailed Implementation

[0098] The specific embodiments of the present invention will now be described in detail with reference to the accompanying drawings, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments.

[0099] Example

[0100] The embodiments given below are provided to facilitate understanding of the present invention and are not intended to limit the scope of the claims in any way.

[0101] The main materials used in the following embodiments include:

[0102] Recombinant type III collagen: prepared and purified according to the method disclosed in Chinese patent application 201310701942.8, and its amino acid sequence is shown in SEQ ID No: 1.

[0103] SEQ ID No: 1

[0104]

[0105] Recombinant type I collagen: prepared and purified according to the method disclosed in Chinese patent application 201310701767.2, and its amino acid sequence is shown in SEQ ID No: 2.

[0106] SEQ ID No: 2

[0107]

[0108] Recombinant human collagen: prepared and purified according to the method disclosed in Chinese patent application 01106757.8, and its amino acid sequence is shown in SEQ ID No: 3.

[0109] SEQ ID No: 3

[0110] GAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAP

[0111] GAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAP

[0112] GSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQ

[0113] GAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAP

[0114] GLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQ

[0115] GAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAP

[0116] GAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAP

[0117] GSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQ

[0118] GAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAP

[0119] GLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQ

[0120] GAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAP

[0121] GAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAP

[0122] GSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQ

[0123] GAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAP

[0124] GLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQ

[0125] GAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAPGAPGSQGAPGLQGAP

[0126] GAPGSQGAPGLQ

[0127] Mini small molecule peptide-1: APG tripeptide, synthesized by a commercial company.

[0128] Mini small molecule peptide-2: LPG tripeptide, synthesized by a commercial company.

[0129] Example 1: Effects of individual components and compositions on the amount of growth factor in mouse fibroblasts

[0130] The following reagents were prepared: type III recombinant collagen solution (25 mg / mL, aqueous solution), APG small molecule peptide solutions 1-5 (0.01 mg / mL, 0.1 mg / mL, 1 mg / mL, 5 mg / mL, 10 mg / mL, aqueous solution), compositions 1-5 (type III recombinant collagen 25 mg / mL + APG small molecule peptides 0.01 mg / mL, 0.1 mg / mL, 1 mg / mL, 5 mg / mL, 10 mg / mL, aqueous solution), commercially available recombinant collagen solution (25 mg / mL, aqueous solution), and 100 mL of purified water.

[0131] The above solution was used to administer the drug to mice in groups via topical application, and the specific steps are as follows:

[0132] After one week of acclimatization, 110 mice weighing 25-30g were selected, and the remaining hair on the back of each mouse was shaved off in a 1.0×1.0cm area. Mice were randomly divided into 11 groups of 10 mice each. The purified water group served as the normal control group, and a commercially available recombinant collagen solution served as the positive control group. The remaining mice were randomly divided into three groups: type III recombinant collagen group, APG small molecule collagen peptide-1 group (0.01mg / mL), APG small molecule collagen peptide-2 group (0.1mg / mL), APG small molecule collagen peptide-3 group (1mg / mL), APG small molecule collagen peptide-4 group (5mg / mL), APG small molecule collagen peptide-5 group (10mg / mL), and combinations 1-5. The mice were treated twice daily (morning and evening) at a dose of 0.2mL / 10g for 4 weeks. Two hours after the last administration, the mice were sacrificed, and skin samples were harvested from the back. Tissue homogenates were prepared for ELISA to detect fibroblast growth factor (β-FGF) levels. Calculate the growth rate of b-FGF in each treatment group.

[0133] β-FGF growth rate = β-FGF level in the treatment group / β-FGF level in the normal group

[0134] Simultaneously, the combination drug index CI = (A × B) / AB was calculated. Where A and B represent the β-FGF growth rates of the type III recombinant collagen group and the APG small molecule collagen peptide-1-5 group, respectively, and AB represents the β-FGF growth rate of the combination 1-5 group. A CI < 1 indicates a synergistic effect; a CI ≤ 0.9 indicates a significant synergistic effect.

[0135] The effects of each single component and the combination on the amount of mouse fibroblast growth factor and the combined drug index are shown in Table 1.

[0136] Table 1

[0137] Group b-FGF (pg / mL) b-FGF growth rate CI normal group 8.3±0.3 - / Positive control group 9.6±0.2 - / Type III recombinant collagen group 10.3±0.4 1.24 / APG-1 Group 9.1±0.1 1.10 / APG-2 Group 10.9±0.3 1.31 / APG-3 Group 15.2±0.4 1.83 / APG-4 21.2±0.3 2.55 / APG-5 group 29.6±0.1 3.57 / Group 1 of the composition 10.5±0.4 1.27 1.08 Composition 2 groups 14.7±0.1 1.77 0.92 Composition 3 groups 23.6±0.1 2.84 0.80 Composition 4 groups 30.4±0.6 3.66 0.87 5 groups of compositions 36.1±0.3 4.35 1.02

[0138] As shown in Table 1, compositions 1-5 of the present invention can increase the amount of fibroblast growth factor in mice. Meanwhile, the combined drug index of compositions 2-4 is all less than 1, indicating that the ratio of recombinant type III collagen to APG small molecule collagen peptides has a synergistic effect when the ratio is between 250:1 and 5:1. In particular, the combined drug index of composition 3 is all less than or equal to 0.8, indicating that the synergistic effect of recombinant type III collagen to APG small molecule collagen peptides is significant when the ratio is between 25:1 and 5:1. In the skin, type III recombinant collagen promotes fibroblast growth and collagen secretion by fibroblasts, thereby maintaining and restoring skin elasticity and playing a role in anti-wrinkle and firming. APG small molecule collagen peptides, mainly as a type of elastin molecule, can promote the growth of dermal fibroblasts. Therefore, the combined effect of the two can greatly increase the number of fibroblasts in the dermal layer of the skin, achieving the effects of increasing skin elasticity and anti-wrinkle and firming.

[0139] Example 2: Effects of individual components and compositions on the thickness of the dermis in mice

[0140] The following reagents were prepared: recombinant type I collagen solution (25 mg / mL, aqueous solution), LPG small molecule peptide solutions 1-5 (0.01 mg / mL, 0.1 mg / mL, 1 mg / mL, 5 mg / mL, 10 mg / mL, aqueous solution), combinations 1-5 (recombinant type I collagen 25 mg / mL + LPG small molecule peptides 0.01 mg / mL, 0.1 mg / mL, 1 mg / mL, 5 mg / mL, 10 mg / mL, aqueous solution), and 100 mL of purified water.

[0141] The mice were administered the above solution in groups, and the specific steps were as follows:

[0142] After one week of acclimatization, 130 mice weighing 25-30g were selected, and the remaining hair on the back of each mouse was shaved off in a 1.0×1.0cm area. Mice were randomly divided into 13 groups of 10 mice each. The purified water group served as the normal control group, the 10% sodium lactate group as the model group, and the remaining mice, after being treated with 10% sodium lactate for 7 days, were randomly divided into the following groups: recombinant type I collagen group, LPG small molecule collagen peptide-1 group (0.01mg / mL), LPG small molecule collagen peptide-2 group (0.1mg / mL), LPG small molecule collagen peptide-3 group (1mg / mL), LPG small molecule collagen peptide-4 group (5mg / mL), LPG small molecule collagen peptide-5 group (10mg / mL), and combinations 1-5. The medication was applied twice daily, morning and evening, at a dose of 0.2mL / 10g, for 4 weeks. Two hours after the last administration, the mice were sacrificed, and skin was excised from their backs to prepare tissue sections. The thickness of the dermis was measured, and the dermal recovery coefficient of each administration group was calculated.

[0143] Dermal recovery coefficient = Dermal thickness of the drug treatment group / Dermal thickness of the model group

[0144] Simultaneously, the combination drug index CI = (A × B) / AB is calculated. Where A and B are the dermal recovery coefficients of the type I recombinant collagen group and the LPG small molecule peptide-1-5 group, respectively, and AB is the dermal recovery coefficient of the combination 1-5 group. CI < 1 indicates a synergistic effect; CI ≤ 0.9 indicates a significant synergistic effect.

[0145] The dermal recovery coefficients of each individual component and the combination drug index of combinations 1-5 are shown in Table 2.

[0146] Table 2

[0147]

[0148] As shown in Table 2, compared with the 10% lactic acid injury group, compositions 1-5 of the present invention were able to restore the dermal thickness of mice. Meanwhile, the combined drug index of compositions 2-4 was less than 1, indicating that the ratio of recombinant type I collagen to LPG small molecule peptides at 250:1 to 5:1 had a synergistic effect in restoring dermal thickness. In particular, the combined drug index of compositions 3 and 4 was less than 0.9, indicating that the ratio of recombinant type I collagen to LPG small molecule peptides at 25:1 to 5:1 had a significant synergistic effect in restoring dermal thickness. In the skin, recombinant type I collagen mainly functions in the form of collagen fibers, providing tissue flexibility and tear resistance, and also promoting epithelial cell growth and skin repair. On the other hand, no reports on the function of LPG small molecule peptides have been found. This invention is the first to discover that LPG small molecule peptides have the function of increasing dermal thickness and promoting skin repair. Therefore, the present invention also provides the use of the LPG small molecule peptides in the preparation of cosmetics for increasing dermal thickness and promoting skin repair. Changes in dermal structure are a major intrinsic physiological cause of skin aging. Studies have shown that the dermis thins as skin ages. The aforementioned research of this invention demonstrates that the combination of recombinant type I collagen and LPG small molecule peptides can synergistically increase the thickness of the dermal layer, thereby achieving a firming and anti-aging effect.

[0149] Example 3: Preparation of face cream

[0150] The composition of the face cream prepared in this embodiment is as follows:

[0151] 10.0 parts by weight of a recombinant collagen-small peptide composition; the recombinant collagen-small peptide composition is composed of the above-mentioned recombinant type I collagen, recombinant type III collagen and mini small molecule recombinant collagen peptide, wherein the weight ratio of the recombinant type III collagen to the mini small molecule recombinant collagen peptide APG is in the range of 10:1 to 100:1, and the weight ratio of the recombinant type I collagen to the mini small molecule recombinant collagen peptide LPG is in the range of 10:1 to 100:1;

[0152] 10.0 parts by weight of recombinant human collagen;

[0153] 3.0 parts by weight of glycerin;

[0154] 1,2-Pentanediol 1.0 part by weight;

[0155] 4.0 parts by weight of isononyl isononanoate;

[0156] Cetearyl alcohol 2.5 parts by weight;

[0157] 2.0 parts by weight of behenol;

[0158] 3.0 parts by weight of squalane;

[0159] 0.5 parts by weight of tocopherol acetate;

[0160] 0.5 parts by weight of 1,2-hexanediol;

[0161] Dipotassium glycyrrhizate, 0.2 parts by weight;

[0162] Carnosine 0.2 parts by weight;

[0163] 4.0 parts by weight of *Pseudolarix amabilis* extract;

[0164] 2.0 parts by weight of cross-linked polymers of acrylates / C10-30 alkanol acrylates;

[0165] Sodium hyaluronate 0.5 parts by weight;

[0166] Butanediol 4.0 parts by weight;

[0167] 0.05 parts by weight of disodium EDTA;

[0168] Acetyl hexapeptide-8 10.0 parts by weight;

[0169] as well as

[0170] Appropriate amount of purified water

[0171] The preparation method includes the following steps:

[0172] (1) Accurately weigh glycerol, 1,2-pentanediol, 1,2-hexanediol, *Euphorbia milii* extract, sodium hyaluronate, butylene glycol, recombinant collagen-peptide composition, recombinant human collagen, and disodium EDTA. Disperse them evenly, mix them with some purified water, add them to the main pot, stir evenly, heat, and keep warm at 85±2℃ for 30 minutes.

[0173] (2) Accurately weigh the isononyl isononanoate, cetearyl alcohol, behenyl alcohol, squalane, and tocopheryl acetate; heat to dissolve, and keep warm at 80-85℃ for 10 min;

[0174] (3) After the heat preservation is completed, add the raw materials mentioned in (2) into the main pot, stir and homogenize until the material is uniform, add the pre-dispersed isononyl isononanoate and hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer into the main pot, and continue to emulsify and stir until the material is uniform.

[0175] (4) Cool to 45℃-50℃, add the dissolved dimethyl glycyrrhizate and carnosine to the mixing bowl, and stir well. Then add acetyl hexapeptide-8 to the mixing bowl, continue stirring, and cool to below 30℃ to obtain the final product. Example 3: Effect of face cream on zebrafish embryos

[0176] The above-mentioned face cream was used to test zebrafish embryos, and the specific steps are as follows:

[0177] Twenty-four zebrafish embryos aged 48 hours were exposed to a 5 g / L solution of the aforementioned face cream, with a blank control group included. After 24 hours of exposure, the embryos were stained with H2DCFDA, and the ROS signal intensity was measured by fluorescence imaging and statistical analysis was performed.

[0178] Wrinkle formation is related to the oxidation and inactivation of proteins, nucleic acids, lipids, and other molecules caused by reactive oxygen species (ROS). In daily life, various factors such as sun exposure and chemical irritants can increase oxidative stress damage to the skin, leading to abnormal skin aging, loss of elasticity, and wrinkles. Figure 1 The results show that the above-mentioned face cream sample achieved a 20% ROS scavenging rate from zebrafish embryos at a test concentration of 5 g / L. This indicates that the sample can significantly remove ROS from zebrafish embryos, demonstrating antioxidant effects and supporting claims of anti-wrinkle and firming efficacy.

[0179] Example 4: Effect of face cream on collagen gene expression in zebrafish embryos

[0180] The above-mentioned face cream was used to test zebrafish embryos, and the specific steps are as follows:

[0181] Zebrafish were randomly selected and placed in 6-well plates, 30 fish per well. Water-soluble samples were administered, with a normal control group included. Each well contained 3 mL of the sample. Three biological replicates were performed. The plates were incubated at 28°C in the dark for 24 h. Total RNA was extracted from the zebrafish in each experimental group, and cDNA was synthesized. The expression of β-actin and the target gene was detected using q-PCR. β-actin was used as an internal control for gene expression. The relative expression level of the target gene RNA was calculated (ΔΔCt = (target gene Ct value in the treatment group - internal control gene Ct value in the treatment group) (target gene Ct value in the control group - internal control gene Ct value in the control group), with an average relative content of 2 - ΔΔCt × 100%.

[0182] Depend on Figure 2 It can be seen that the expression of genes related to type I and type III collagen was significantly increased compared with the control group, indicating that the use of face cream will significantly increase the content of type I and type III collagen in zebrafish embryos.

[0183] Example 5: The effect of face cream on skin firmness, elasticity, and smoothness.

[0184] The above-mentioned face cream was used to conduct an application test on volunteers, and the specific steps are as follows:

[0185] Thirty-two volunteers aged 25-45 were recruited. After cleansing, the initial skin firmness of the left and right sides of their faces was tested as a baseline. The skincare product was used twice daily, morning and evening, for four weeks, with 3g used each time. Skin firmness was assessed after four weeks of product use, and the improvement rate was calculated.

[0186] Improvement rate = Base value - (Measurement value after four weeks of product use) / (Measurement value after four weeks of product use)

[0187] Table 3. Effects of face cream on skin firmness, elasticity, and smoothness.

[0188]

[0189]

[0190] As shown in Table 3, after four weeks of continuous use of the above products, the subjects' skin firmness, elasticity, and smoothness values ​​were significantly different from the baseline values. Skin firmness increased by 10.58%, skin elasticity increased by 13.33%, and skin smoothness increased by 11.34%.

[0191] Example 6: The effect of face cream on wrinkles on human skin

[0192] The above-mentioned face cream was used to conduct an application test on volunteers, and the specific steps are as follows:

[0193] Thirty-two volunteers aged 25-45 were recruited. After cleansing, the initial skin firmness of the left and right sides of the face was tested as a baseline. The skincare product was used twice daily, morning and evening, for four weeks, with 3g used each time. The area of ​​wrinkles on the skin was measured after four weeks of product use, and the improvement rate of skin condition after four weeks was calculated.

[0194] Improvement rate = Base value - (Measurement value after four weeks of product use) / (Measurement value after four weeks of product use)

[0195] Table 4. The effect of face cream on wrinkles on human skin.

[0196]

[0197] As shown in Table 4, after four weeks of continuous use of the above products, the Visia-CR cheek texture area of ​​the subjects showed a highly significant difference compared with the baseline value, with a decrease rate of 25.92% in cheek skin texture area and a decrease rate of 22.46% in texture area percentage.

[0198] Example 7: Preparation of Eye Cream

[0199] The composition of the eye cream prepared in this embodiment is as follows:

[0200] 12.0 parts by weight of a recombinant collagen-small peptide composition; the recombinant collagen-small peptide composition is composed of the above-mentioned recombinant type I collagen, recombinant type III collagen and mini small molecule recombinant collagen peptide, wherein the weight ratio of the recombinant type III collagen to the mini small molecule recombinant collagen peptide APG is in the range of 10:1 to 100:1, and the weight ratio of the recombinant type I collagen to the mini small molecule recombinant collagen peptide LPG is in the range of 10:1 to 100:1;

[0201] 10.0 parts by weight of recombinant human collagen;

[0202] 5.0 parts by weight of glycerin;

[0203] 1,2-Pentanediol 1.0 part by weight;

[0204] 4.0 parts by weight of isononyl isononanoate;

[0205] Cetearyl alcohol 2.0 parts by weight;

[0206] 1.5 parts by weight of behenol;

[0207] 4.0 parts by weight of squalane;

[0208] 0.3 parts by weight of tocopherol acetate;

[0209] 0.5 parts by weight of 1,2-hexanediol;

[0210] Dipotassium glycyrrhizate, 0.2 parts by weight;

[0211] Carnosine 0.2 parts by weight;

[0212] 3.0 parts by weight of *Pseudolarix amabilis* extract;

[0213] 2.0 parts by weight of cross-linked polymers of acrylates / C10-30 alkanol acrylates;

[0214] Sodium hyaluronate 1.0 part by weight;

[0215] Butanediol 4.0 parts by weight;

[0216] 0.05 parts by weight of disodium EDTA;

[0217] Chlorella extract 5.0 parts by weight;

[0218] Mulberry extract 1.0 part by weight;

[0219] Acetyl hexapeptide-8 15.0 parts by weight;

[0220] And, an appropriate amount of purified water.

[0221] The preparation method includes the following steps:

[0222] (1) Accurately weigh glycerol, 1,2-pentanediol, 1,2-hexanediol, *Euphorbia milii* extract, sodium hyaluronate, butylene glycol, recombinant collagen-peptide composition, recombinant human collagen, and disodium EDTA. Disperse them evenly, mix them with some purified water, add them to the main pot, stir evenly, heat, and keep warm at 85±2℃ for 30 minutes.

[0223] (2) Accurately weigh the isononyl isononanoate, cetearyl alcohol, behenyl alcohol, squalane, and tocopheryl acetate; heat to dissolve, and keep warm at 80-85℃ for 10 min;

[0224] (3) After the heat preservation is completed, add the raw materials mentioned in (2) into the main pot, stir and homogenize until the material is uniform, add the pre-dispersed isononyl isononanoate and hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer into the main pot, and continue to emulsify and stir until the material is uniform.

[0225] (4) Cool down to 45℃-50℃, add the dissolved dimethyl glycyrrhizate and carnosine to the main pot, and stir well. Then add acetyl hexapeptide-8, chlorella extract and mulberry extract to the main pot in sequence, and continue stirring and cooling down to below 30℃ to obtain the final product.

[0226] Example 8: The effect of eye cream on wrinkles under the eyes

[0227] The above-mentioned eye cream was used to conduct an application test on volunteers, and the specific steps are as follows:

[0228] Thirty-two volunteers aged 25-45 were recruited. After cleansing, the initial skin firmness of the left and right sides of the face was tested as a baseline. The skincare product was used twice daily, morning and evening, for four weeks, with 1g used each time. The area of ​​wrinkles under the eyes was measured after four weeks of product use, and the improvement rate of skin condition after four weeks was calculated.

[0229] Improvement rate = Base value - (Measurement value after four weeks of product use) / (Measurement value after four weeks of product use)

[0230] Table 5. Effects of eye cream on wrinkles under the eyes.

[0231]

[0232] As shown in Table 5, after four weeks of continuous use of the above products, the Visia-CR under-eye wrinkle area of ​​the subjects was significantly different from the baseline value, with a reduction rate of 19.72% and a reduction rate of 20.35% in the proportion of under-eye wrinkle area.

[0233] Example 9: Preparation of a facial mask

[0234] The composition of the topical mask prepared in this embodiment is as follows:

[0235] 12.0 parts by weight of a recombinant collagen-small peptide composition; the recombinant collagen-small peptide composition is composed of the above-mentioned recombinant type I collagen, recombinant type III collagen and mini small molecule recombinant collagen peptide, wherein the weight ratio of the recombinant type III collagen to the mini small molecule recombinant collagen peptide APG is in the range of 10:1 to 100:1, and the weight ratio of the recombinant type I collagen to the mini small molecule recombinant collagen peptide LPG is in the range of 10:1 to 100:1;

[0236] 1-20 parts by weight of the recombinant collagen-peptide composition;

[0237] 15 parts by weight of recombinant human collagen;

[0238] 8 parts by weight of glycerin;

[0239] 1 part by weight of 1,2-pentanediol;

[0240] 6 parts by weight of isononyl isononanoate;

[0241] 4 parts by weight of cetearyl alcohol;

[0242] Butanediol 4 parts by weight;

[0243] 1 part by weight of sodium hyaluronate;

[0244] 3 parts by weight of behenol;

[0245] 5 parts by weight of squalane;

[0246] 0.5 parts by weight of tocopherol acetate;

[0247] 0.9 parts by weight of hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer;

[0248] Mulberry extract, 0.3 parts by weight;

[0249] And, an appropriate amount of purified water.

[0250] The preparation method includes the following steps:

[0251] (1) Accurately weigh glycerol, 1,2-pentanediol, sodium hyaluronate, butylene glycol, recombinant collagen-peptide composition, and recombinant human collagen, disperse them evenly, mix them with some purified water, add them to the main pot, stir evenly, heat, and keep warm at 85±2℃ for 30min.

[0252] (2) Accurately weigh the isononyl isononanoate, cetearyl alcohol, behenyl alcohol, squalane, and tocopheryl acetate; heat to dissolve, and keep warm at 80-85℃ for 10 min;

[0253] (3) After the heat preservation is completed, add the raw materials mentioned in (2) into the main pot, stir and homogenize until the material is uniform, add the pre-dispersed isononyl isononanoate and hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer into the main pot, and continue to emulsify and stir until the material is uniform.

[0254] (4) Cool down to 45℃-50℃, add the dissolved mulberry extract to the main pot in sequence, continue stirring and cool down to below 30℃ to obtain the product.

[0255] The above-mentioned face mask was used to conduct an application test on volunteers, and the specific steps are as follows:

[0256] Thirty-two volunteers aged 25-45 were recruited. After cleansing, the initial skin firmness of the left and right sides of the face was tested as a baseline. The skincare product was used twice daily, morning and evening, for four weeks, with 3g used each time. The area of ​​wrinkles on the skin was measured after four weeks of product use, and the improvement rate of skin condition after four weeks was calculated.

[0257] Improvement rate = Base value - (Measurement value after four weeks of product use) / (Measurement value after four weeks of product use)

[0258] Table 6. Effects of applying face masks on wrinkles on human facial skin.

[0259]

[0260] As shown in Table 6, after four weeks of continuous use of the above products, the Visia-CR cheek wrinkle area of ​​the subjects was significantly different from the baseline value, with a cheek wrinkle area reduction rate of 27.72% and a cheek wrinkle area percentage reduction rate of 20.85%.

Claims

1. A cosmetic comprising a recombinant collagen-small peptide composition, and one or more selected from the group consisting of a recombinant human-like collagen, a skin conditioner, an emulsifier, an antioxidant, an emollient, a humectant, a thickening agent, a complexing agent, a solvent; wherein, the recombinant collagen-small peptide composition consists of a recombinant collagen type III, a recombinant collagen type I, an APG tripeptide, and an LPG tripeptide; the weight ratio of the recombinant collagen type III and the APG tripeptide in the recombinant collagen-small peptide composition is 250: 1 to 5: 1, and / or, the weight ratio of the recombinant collagen type I and the LPG tripeptide is 250: 1 to 5: 1; the amino acid sequence of the recombinant collagen type III is shown in SEQ ID No: 1; the amino acid sequence of the recombinant collagen type I is shown in SEQ ID No: 2; and the amino acid sequence of the recombinant human-like collagen is shown in SEQ ID No:

3. The weight ratio of the recombinant collagen type III and the APG tripeptide is 25: 1 to 5:

1. The weight ratio of the recombinant collagen type I and the LPG tripeptide is 25: 1 to 5:

1. The skin conditioner is a combination of one or more selected from the group consisting of acetyl hexapeptide-8, 1, 3-propanediol, chlorella extract, and mulberry extract. The emulsifier is one or more selected from the group consisting of cetostearyl alcohol, behenyl alcohol, trisbehenin, and arachidyl alcohol. The antioxidant is selected from the group consisting of p-hydroxyacetophenone.

2. The cosmetic according to claim 1, wherein The emollient is one or more selected from the group consisting of caprylic / capric triglyceride, isononyl isononanoate, dimethicone, squalane, ethyl ethylglycerin, and the like.

3. The cosmetic according to claim 1, wherein The humectant is one or more selected from the group consisting of glycerin, soluble collagen, 1, 2-pentanediol, 1, 2-hexanediol, caprylyl glycol, sodium hyaluronate, and the like.

4. The cosmetic according to claim 1, wherein The thickening agent is one or more selected from the group consisting of hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer, acrylates / C 10-30 alkyl acrylate crosspolymer, and the like.

5. The cosmetic according to claim 1, wherein The complexing agent is selected from the group consisting of disodium EDTA and phytic acid.

6. The cosmetic according to claim 1, wherein 11.The cosmetic according to claim 1, which is a skin cream, a skin milk, a face cream, an eye cream, or a spread mask.

7. The cosmetic according to claim 1, wherein 12.The cosmetic according to claim 1, which is a face cream comprising: the recombinant collagen-small peptide composition 1 to 20 parts by weight; the recombinant human-like collagen 1 to 20 parts by weight; glycerin 1 to 10 parts by weight; 1, 2-pentanediol 1 to 3 parts by weight; isononyl isononanoate 1 to 10 parts by weight; cetostearyl alcohol 1 to 5 parts by weight; behenyl alcohol 1 to 5 parts by weight; squalane 1 to 10 parts by weight; tocopheryl acetate 0.5 to 5 parts by weight; 1, 2-hexanediol 0.1 to 3 parts by weight; dipotassium glycyrrhizinate 0.1 to 1 parts by weight; carnosine 0.1 to 2 parts by weight; chlorella extract 1 to 5 parts by weight; acrylates / C 10-30 alkyl acrylate crosspolymer 0.1 to 3 parts by weight; sodium hyaluronate 0.1 to 3 parts by weight; butylene glycol 1 to 10 parts by weight; disodium EDTA 0.01 to 1 parts by weight; and acetyl hexapeptide-8 0.1 to 10 parts by weight.

8. The cosmetic according to claim 1, wherein ​ 9. The cosmetic according to claim 1, wherein ​ 10. The cosmetic according to claim 1, wherein ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ tocopheryl acetate 0.01 to 5 parts by weight; and purified water in an appropriate amount.

13. The cosmetic according to claim 1, which is a pack, comprising: the recombinant collagen-small peptide composition 1 to 20 parts by weight; recombinant human-like collagen 1 to 20 parts by weight; glycerin 1 to 10 parts by weight; 1,2-pentanediol 1 to 3 parts by weight; isononyl isononanoate 1 to 10 parts by weight; cetostearyl alcohol 1 to 5 parts by weight; butylene glycol 1 to 10 parts by weight; sodium hyaluronate 0.1 to 2 parts by weight; behenyl alcohol 1 to 5 parts by weight; squalane 1 to 10 parts by weight; tocopheryl acetate 0.5 to 5 parts by weight; hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer 0.1 to 2 parts by weight; mulberry extract 0.1 to 10 parts by weight; and purified water in an appropriate amount.

14. The cosmetic according to claim 1, which is an eye cream, comprising: the recombinant collagen-small peptide composition 1 to 20 parts by weight; recombinant human-like collagen 1 to 20 parts by weight; glycerin 1 to 10 parts by weight; 1,2-pentanediol 1 to 3 parts by weight; isononyl isononanoate 1 to 10 parts by weight; cetostearyl alcohol 1 to 5 parts by weight; behenyl alcohol 1 to 5 parts by weight; squalane 1 to 10 parts by weight; tocopheryl acetate 0.1 to 5 parts by weight; 1,2-hexanediol 0.1 to 5 parts by weight; dipotassium glycyrrhizinate 0.1 to 2 parts by weight; carnosine 0.1 to 2 parts by weight; calophyllum inophyllum extract 1 to 10 parts by weight; acrylate / C10-30 alkyl acrylate crosspolymer 1 to 5 parts by weight; sodium hyaluronate 0.1 to 3 parts by weight; butylene glycol 1 to 10 parts by weight; disodium EDTA 0.01 to 1 parts by weight; chlorella extract 1 to 10 parts by weight; mulberry extract 0.1 to 5 parts by weight; acetyl hexapeptide-8 1 to 20 parts by weight; tocopheryl acetate 0.01 to 5 parts by weight; and purified water in an appropriate amount.

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