A peptide-containing skin care composition with anti-wrinkle firming efficacy

Abstract

The application discloses a peptide-containing skin care composition with anti-wrinkle and firming effects, which solves the technical contradiction between high cost and low stability by compounding specific sequence type I collagen peptide and chitosan acetate. In view of the high cost of solid-phase synthesis of human collagen peptide, the synergistic effect of the two is utilized, so that the collagen peptide can play a significant effect at a very low concentration, and the raw material cost is greatly reduced. In view of the problem that chitosan acetate is prone to causing formula yellowing and poor stability at a high concentration, the present scheme controls the concentration of chitosan acetate in a low-concentration safe range, avoids yellowing, and improves the shelf life stability. The low-concentration compounding path realizes the best balance between cost control and formula stability while retaining biological activity, and has both economy and safety.

Description

Technical Field

[0001] This invention belongs to the field of peptide-containing biological composition technology, and relates to a peptide-containing skin care composition with anti-wrinkle and firming effects. Background Technology

[0002] The most obvious signs of skin aging are wrinkles and sagging, which stem from the decline in the synthesis of hyaluronic acid and collagen in the extracellular matrix (ECM) with age, leading to a loss of skin support and elasticity. Therefore, enhancing ECM synthesis through exogenous supplementation or endogenous activation is a key technological approach in the field of anti-wrinkle and firming. In this area, peptide-containing biosynthetic compositions have become a research hotspot due to their excellent biocompatibility and targeting properties.

[0003] In existing technologies, collagen peptides, as typical bioactive peptides, have been proven to promote cell proliferation and hyaluronic acid synthesis. Studies have shown that specific sequences rich in proline-hydroxyproline (Pro-Hyp) dipeptides are key functional groups for their efficacy. For example, Chinese patent application CN114404605B discloses a specific polypeptide sequence derived from human type I collagen (Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2), which exhibits significant anti-wrinkle potential. However, such high-purity, specific-sequence functional peptides are currently mainly prepared using solid-phase synthesis. Due to the cumbersome synthesis process and limited yield, the raw material cost is extremely high, severely restricting their large-scale application and widespread use in peptide-containing skincare products.

[0004] On the other hand, to synergistically enhance the efficacy of peptide compositions, existing technologies often attempt to combine them with small-molecule active ingredients such as acetylglucosamine. Although acetylglucosamine can help promote collagen synthesis by fibroblasts, its biological activity is relatively weak, and it usually requires a high concentration of more than 2% to be effective. However, in cosmetic composition systems, high concentrations of amino sugars are highly susceptible to Maillard reactions with other ingredients in the formulation, leading to severe yellowing of the product over time and significantly reducing the stability of the formulation.

[0005] In summary, the field of peptide-containing anti-aging compositions currently faces significant technical bottlenecks: on the one hand, high-efficiency specific-sequence collagen peptides are limited by synthesis processes, resulting in high costs; on the other hand, commonly used synergistic ingredients exhibit stability defects at high concentrations. How to overcome the dual limitations of peptide preparation cost and formulation stability, and develop a novel, low-cost, highly stable peptide-containing skincare composition with synergistic effects, is a technical problem urgently needing to be solved by those skilled in the art. Summary of the Invention

[0006] To address the problems existing in the prior art, the present invention provides a peptide-containing skin care composition with anti-wrinkle and firming effects, thereby solving the dual technical problems faced by existing peptide-containing anti-aging compositions: high cost due to solid-phase synthesis process of high-efficiency specific sequence collagen peptides, and poor formula stability due to the need for high concentration of synergistic ingredient acetyl glucosamine which easily triggers Maillard reaction.

[0007] This invention is achieved through the following technical solution: A peptide-containing skincare composition with anti-wrinkle and firming effects includes type I collagen peptides and acetyl glucosamine, wherein the amino acid sequence structure of the type I collagen peptides is Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2.

[0008] Preferably, the mass ratio of the type I collagen peptide to acetyl glucosamine is 1:(1~50).

[0009] Preferably, the mass ratio of the type I collagen peptide to acetyl glucosamine is 1:(10~20).

[0010] Preferably, the molecular weight of the type I collagen peptide is 1521 Da.

[0011] Preferably, the mass fraction of hydroxyproline in the type I collagen peptide is greater than 30%.

[0012] Preferably, the purity of the acetyl glucosamine is 98%~99%.

[0013] The above-mentioned peptide-containing skincare composition with anti-wrinkle and firming effects is used in cosmetics.

[0014] Preferably, it is added to cosmetics at a concentration of 0.1% to 1% by mass.

[0015] Preferably, it is added to cosmetics at a concentration of 0.5% by weight.

[0016] Preferably, the cosmetics include any one of the following: facial mask, lotion, serum, face cream, essence lotion, toner, lotion, moisturizing lotion, eye cream, facial cleanser, sunscreen, nourishing cream, massage cream, anti-wrinkle agent, acne treatment agent, after-sun preparation, and whitening agent.

[0017] Compared with the prior art, the present invention has the following beneficial technical effects: This invention discloses a peptide-containing skincare composition with anti-wrinkle and firming effects. This solution resolves the technical contradiction of high cost and low stability in existing technologies by compounding type I collagen peptides with a specific sequence and acetyl glucosamine. Addressing the issue of high costs associated with solid-phase synthesis of human collagen peptides rich in proline-hydroxyproline structures, this invention utilizes the synergistic effect of both in promoting extracellular matrix synthesis. This allows type I collagen peptides to exert significant effects at extremely low concentrations, thereby greatly reducing the amount of expensive active ingredients used and directly lowering raw material costs. Regarding the issue of yellowing and poor stability of acetyl glucosamine at high concentrations, this invention controls the concentration of acetyl glucosamine within a low and safe range while ensuring efficacy, avoiding yellowing caused by high concentrations and significantly improving the product's shelf-life stability. This technical approach of achieving synergistic effects through low-concentration compounding not only preserves the bioactivity of both components but also finds the optimal balance between cost control and formula stability.

[0018] Furthermore, the mass ratio of the type I collagen peptide to acetyl glucosamine is 1:(1~50). This ratio range ensures that acetyl glucosamine can play a precursor role in promoting hyaluronic acid synthesis in the formulation, while also ensuring that the collagen peptide provides sufficient amino acid signals.

[0019] Furthermore, the mass ratio of the type I collagen peptide to acetyl glucosamine is 1:(10~20), which allows the two to produce the best synergistic effect when combined. Detailed Implementation

[0020] To enable those skilled in the art to understand the features and effects of the present invention, the terms and expressions used in the specification and claims are explained and defined in general below. Unless otherwise specified, all technical and scientific terms used herein have the ordinary meaning understood by those skilled in the art regarding the present invention, and in case of conflict, the definitions in this specification shall prevail.

[0021] The theories or mechanisms described and disclosed herein, whether right or wrong, should not in any way limit the scope of the invention, that is, the contents of the invention can be implemented without being limited by any particular theory or mechanism.

[0022] In this document, all features defined by numerical ranges or percentage ranges, such as numerical values, quantities, contents, and concentrations, are for the sake of brevity and convenience only. Accordingly, descriptions of numerical ranges or percentage ranges should be considered as covering and specifically disclosing all possible sub-ranges and individual numerical values ​​(including integers and fractions) within those ranges.

[0023] In this article, unless otherwise specified, “contains,” “includes,” “containing,” “has,” or similar terms cover the meanings of “composed of” and “mainly composed of,” for example, “A contains a” covers the meanings of “A contains a and others” and “A contains only a.”

[0024] For the sake of brevity, not all possible combinations of the technical features in each implementation scheme or embodiment are described herein. Therefore, as long as there is no contradiction in the combination of these technical features, the technical features in each implementation scheme or embodiment can be combined arbitrarily, and all possible combinations should be considered within the scope of this specification.

[0025] This invention provides a peptide-containing skincare composition with anti-wrinkle and firming effects, comprising type I collagen peptides and acetyl glucosamine, wherein the amino acid sequence structure of the type I collagen peptides is Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2.

[0026] The mass ratio of type I collagen peptide to acetyl glucosamine is 1:(1~50), which can be 1:5, 1:10 or 1:20, preferably 1:(10~20), and most preferably 1:10.

[0027] This type I collagen peptide is a biomimetic short peptide with specific chemical modifications. Its amino acid sequence is Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2. This sequence consists of 14 amino acid residues, with extremely high total content of glycine (G), proline (P), and hydroxyproline (Hyp). In addition, the sequence also contains phenylalanine (F), glutamic acid (E), and arginine (R). Furthermore, the N-terminus of the peptide chain is modified with acetylation (Ac-), and the C-terminus is blocked with amidation (-NH2). This specific sequence arrangement and terminal modification structure not only highly mimics the bioactive center of natural human type I collagen but also significantly enhances the enzymatic stability and transdermal absorption capacity of the peptide chain, thus endowing it with excellent biological activity.

[0028] This invention utilizes a combination of collagen peptides with specific sequences and acetyl glucosamine to leverage their synergistic effect in promoting ECM synthesis in skin cells, thereby achieving the beneficial effects of reducing costs, enhancing anti-wrinkle and firming effects, and improving formula stability.

[0029] The type I collagen peptide in this invention is rich in proline-hydroxyproline structure. This collagen peptide sequence is derived from human type I collagen. The molecular weight of the type I collagen peptide is 1521 Daltons (Da), and the hydroxyproline content is greater than 30%, meaning the mass fraction of hydroxyproline in the type I collagen peptide is greater than 30%. This type I collagen peptide was purchased from Future Polypeptide Biotechnology (Yantai) Co., Ltd.

[0030] The purity of acetyl glucosamine in this invention is 98%~99%.

[0031] Meanwhile, the present invention also discloses the application of the above-mentioned skin care composition in whitening products. Specifically, the skin care composition is added to cosmetics at a mass content of 0.1% to 1%, preferably at a mass content of 0.5%.

[0032] The cosmetics include any one of the following: facial masks, lotions, serums, face creams, essence lotions, toners, lotions, moisturizing lotions, eye creams, facial cleansers, sunscreens, nourishing creams, massage creams, anti-wrinkle agents, acne treatments, after-sun preparations, and whitening agents.

[0033] The collagen peptide used in this invention is a specific sequence of human type I collagen peptide, synthesized via solid-phase synthesis. This sequence is rich in proline-hydroxyproline structures. Hyaluronic acid synthase 2 (HAS2) is one of the key enzymes for the synthesis of hyaluronic acid (HA) in the human body. It mainly catalyzes the production of high molecular weight hyaluronic acid on the cell membrane and plays a central role in maintaining skin elasticity, skin plumpness, and cell signaling. Studies have shown that the proline-hydroxyproline dipeptide structure enhances HA synthesis by regulating the expression of HAS2 mRNA in fibroblasts, thereby promoting fibroblast proliferation and collagen synthesis. Other studies have shown that acetylglucosamine, as a HA precursor, can enhance cellular HA synthesis and promote collagen synthesis by increasing type I collagen mRNA expression, but it has no regulatory effect on cellular HAS2 mRNA expression. By combining this type I collagen peptide with acetyl glucosamine, the resulting composition synergistically promotes HA synthesis in the ECM by targeting two important targets: HAS2 mRNA expression and the provision of essential HA precursors. HA synthesis further promotes collagen synthesis in fibroblasts, thereby increasing skin tissue volume, effectively improving wrinkles and sagging, and achieving anti-wrinkle and firming effects. Furthermore, the collagen peptide and acetyl glucosamine in this invention achieve the effects of using them alone at lower concentrations, while at higher concentrations, thus significantly reducing costs and improving formulation stability. In summary, this invention has the advantages of low cost, good anti-wrinkle and firming effects, and good stability.

[0034] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0035] The following examples use instruments and equipment conventional in the art. Experimental methods in the following examples, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. All raw materials used in the following examples are conventional commercially available products with specifications conventional in the art. In this specification and the following examples, unless otherwise specified, "%" refers to weight percentage, "parts" refers to parts by weight, and "ratio" refers to weight proportion.

[0036] Example 1 This embodiment provides a peptide-containing skincare composition with anti-wrinkle and firming effects, comprising type I collagen peptides and acetyl glucosamine, wherein the amino acid sequence structure of the type I collagen peptides is Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2. The mass ratio of the type I collagen peptides to acetyl glucosamine is 1:1.

[0037] Example 2 This embodiment provides a peptide-containing skincare composition with anti-wrinkle and firming effects, comprising type I collagen peptides and acetyl glucosamine, wherein the amino acid sequence structure of the type I collagen peptides is Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2. The mass ratio of the type I collagen peptides to acetyl glucosamine is 1:5.

[0038] Example 3 This embodiment provides a peptide-containing skincare composition with anti-wrinkle and firming effects, comprising type I collagen peptides and acetyl glucosamine, wherein the amino acid sequence structure of the type I collagen peptides is Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2. The mass ratio of the type I collagen peptides to acetyl glucosamine is 1:10.

[0039] Example 4 This embodiment provides a peptide-containing skincare composition with anti-wrinkle and firming effects, comprising type I collagen peptides and acetyl glucosamine, wherein the amino acid sequence structure of the type I collagen peptides is Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2. The mass ratio of the type I collagen peptides to acetyl glucosamine is 1:20.

[0040] Example 5 This embodiment provides a peptide-containing skincare composition with anti-wrinkle and firming effects, comprising type I collagen peptides and acetyl glucosamine, wherein the amino acid sequence structure of the type I collagen peptides is Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2. The mass ratio of the type I collagen peptides to acetyl glucosamine is 1:30.

[0041] Example 6 This embodiment provides a peptide-containing skincare composition with anti-wrinkle and firming effects, comprising type I collagen peptides and acetyl glucosamine, wherein the amino acid sequence structure of the type I collagen peptides is Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2. The mass ratio of the type I collagen peptides to acetyl glucosamine is 1:50.

[0042] To verify the effectiveness of the technical solution in this invention, the following tests were conducted: I. Experimental Principle Fibroblasts are important functional cells in the dermis. The hyaluronic acid and collagen they synthesize and secrete are the most important structural components of the dermis, closely related to skin wrinkles, elasticity, and water retention. This experiment used human fibroblasts as the experimental subject to test the synthesis of hyaluronic acid and type I collagen in fibroblasts under different sample conditions, and used these two indicators to evaluate and compare the anti-wrinkle and firming effects of different samples.

[0043] II. Experimental Objective: This study investigates whether the peptide-containing skincare composition of the present invention, which has anti-wrinkle and firming effects, actually has anti-wrinkle and firming effects, and confirms whether the combination of type I collagen peptides and acetyl glucosamine can have a synergistic effect, thereby reducing the amount of each compounding component used, and achieving the goals of good efficacy, low cost and good stability.

[0044] III. Main Experimental Equipment and Reagents Main equipment: CO2 incubator, clean bench, microplate reader, upright microscope; Main reagents: Dulbecco's Modified Eagle Medium (DMEM) culture medium, fetal bovine serum, phosphate-buffered saline (PBS), Collagen Type I Enzyme-Linked Immunosorbent Assay (Collagen I ELISA) kit, and Hyaluronic Acid Enzyme-Linked Immunosorbent Assay (HAELISA) kit.

[0045] In this invention, the basic cell culture medium is Dulbecco's Modified Eagle Medium (DMEM), which serves as the basic nutrient solution for in vitro cell culture, providing essential nutrients such as amino acids, vitamins, inorganic salts, and glucose required for cell growth. Fetal bovine serum (FBS) is used as a supplementary additive to the cell culture medium, providing growth factors, hormones, and adhesion factors required for cell proliferation and adhesion. Combined with DMEM, it forms a complete culture medium to maintain normal cell physiological activity. Phosphate-buffered saline (PBS) is used for cell washing and environmental maintenance during experiments. It has an osmotic pressure and pH similar to human body fluids, used to remove residual culture medium components or metabolic waste from the cell surface while maintaining the integrity of cell morphology and structure. The Collagen Type I Enzyme-Linked Immunosorbent Assay (Collagen I...) is used. The ELISA kit is used to quantitatively detect the expression level of type I collagen in cell culture supernatant to assess the ability of the test sample to promote collagen synthesis in the extracellular matrix; the hyaluronic acid enzyme-linked immunosorbent assay (HA ELISA) kit is used to quantitatively detect the content of hyaluronic acid in cell culture supernatant to assess the efficacy of the test sample in promoting hyaluronic acid synthesis in cells and improving skin hydration.

[0046] IV. Experimental Procedure Vaccination: 2 x 10 5 Inoculate cells into 6-well plates at a seeding density of 1 cell per well and incubate overnight in an incubator (37°C, 5% CO2).

[0047] Collagen and hyaluronic acid content determination: According to the test protocol in Table 1, when the cell layering rate in the 6-well plates reached 40%–60%, the culture medium was replaced with a serum- and growth factor-free medium. After 24 hours, the cells were administered to groups, with 2 mL of the medium per well and 3 replicates per group. The cells were incubated in an incubator (37°C, 5% CO2) for 24 hours. The cell culture supernatant was collected in microcentrifuge tubes.

[0048] ELISA assay: Perform the assay according to the instructions for use of the Collagen I ELISA kit.

[0049] Results and statistical analysis: Results are expressed as Mean ± SD. t-tests were used for comparisons between groups.

[0050] Table 1 Test scheme for verifying the technical effect of the present invention

[0051] V. Results Analysis Table 2 shows the synergistic effect of the combination of type I collagen peptides rich in Pro-Hyp structure and acetyl glucosamine on the synthesis of HA and Collagen-I in fibroblasts. Table 2. Effects of the technical solutions in this invention on HA and Collagen-I synthesis in fibroblasts.

[0052] Table 2 details the effects of different ratios of type I collagen peptides and acetyl glucosamine compound compositions on the secretion of hyaluronic acid (HA) and type I collagen (Collagen-I) by fibroblasts. The data in Table 2 show that the compound compositions described in this invention exhibit significant promoting effects across a wide range of ratios, and demonstrate excellent synergistic effects within specific intervals.

[0053] As shown in Table 2, the single component (① type I collagen peptide or ② acetylglucosamine) in each example significantly increased the synthesis of HA and Collagen-I compared to the blank control group (P<0.01). For example, in Examples 1-3, using type I collagen peptide alone increased the HA content by approximately 80% and Collagen-I by approximately 117%; using acetylglucosamine alone also increased HA and Collagen-I to varying degrees (the increase rate varied from 21% to 57% depending on the concentration). This confirms that the two components each possess independent biological activity.

[0054] Further observation of the data of the compound group (①+②) reveals that with the increase of the proportion of acetylglucosamine, the compound composition of the present invention exhibits a significant promoting effect in a wide range of proportions, and shows an excellent synergistic effect in a specific range.

[0055] When the mass ratio is in the range of 1:10 to 1:20 (as in Examples 3 and 4), the compound composition exhibits excellent synergistic effects. Taking Example 3 (1:10) as an example, its HA synthesis enhancement rate is as high as 209.61%, significantly higher than the sum of the effects of the single components (the sum of the enhancement rates of the single components is about 137%, while the compound reaches 209%), indicating that at this ratio, the two active ingredients have a complementary and enhancing mechanism in promoting extracellular matrix synthesis, which can maximize the activation of fibroblast synthesis capacity. Even at a 1:50 ratio with a very high proportion of acetylglucosamine (Example 6), the compound composition can still maintain an enhancement rate of over 100%, which is better than the effect of single collagen peptides, indicating that the composition has good formulation tolerance. Statistical analysis was performed between the groups in the table. "SD" represents the standard deviation, "*" indicates a significant difference, and **P < 0.01.

[0056] Existing technologies such as collagen peptides are effective but expensive, while acetylglucosamine is inexpensive but unstable at high concentrations. This invention combines the two in a specific ratio, utilizing a synergistic effect to achieve or even surpass the effects of using either at high concentrations with lower dosages (reduced collagen peptide dosage and acetylglucosamine concentration below 2%), thus simultaneously overcoming the challenges of high cost and instability. This approach demonstrates the existence of the synergistic effect through detailed cell experiments (experimental examples).

[0057] This invention combines type I collagen peptides with acetyl glucosamine, resulting in a significant synergistic effect. As shown in Table 2, at a 1:10 ratio, the combined group significantly increased the synthesis of hyaluronic acid (HA) by 209.61%, far exceeding the sum of the single collagen peptide group (80.24%) and the single acetyl glucosamine group (57.53%) (137.77%). Similarly, the combined group significantly increased the synthesis of type I collagen by 183.46%, also significantly exceeding the sum of the two components (152.88%). These experimental results demonstrate that the combined composition is significantly more effective than the sum of the effects of using each component alone in promoting collagen and hyaluronic acid synthesis, and this difference is highly statistically significant (**P<0.01). This invention has produced unexpected technical benefits.

[0058] This invention focuses on the synergistic effect of specific sequence collagen peptides and acetyl glucosamine, rather than the simple superposition of single components. Through this specific combination, the invention achieves a three-in-one technical effect of cost reduction, efficiency improvement, and quality stabilization: on the one hand, it significantly enhances the anti-wrinkle and firming efficacy of the product, achieving technological efficiency improvement; on the other hand, by optimizing the formula to reduce the amount of expensive components and avoiding the addition of high-concentration unstable components, it directly reduces raw material costs and significantly improves the shelf-life stability of the product.

[0059] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A peptide-containing skincare composition with anti-wrinkle and firming effects, characterized in that, It includes type I collagen peptides and acetyl glucosamine, wherein the amino acid sequence structure of type I collagen peptides is Ac-GP-Hyp-GF-Hyp-GERGP-Hyp-GP-Hyp-NH2; The mass ratio of type I collagen peptide to acetyl glucosamine is 1:(1~50).

2. The peptide-containing skincare composition with anti-wrinkle and firming effects according to claim 1, characterized in that, The mass ratio of type I collagen peptide to acetyl glucosamine is 1:(10~20).

3. The peptide-containing skincare composition with anti-wrinkle and firming effects according to claim 1, characterized in that, The molecular weight of the type I collagen peptide is 1521 Da.

4. The peptide-containing skincare composition with anti-wrinkle and firming effects according to claim 1, characterized in that, The mass fraction of hydroxyproline in the type I collagen peptide is greater than 30%.

5. The peptide-containing skincare composition with anti-wrinkle and firming effects according to claim 1, characterized in that, The purity of the acetyl glucosamine is 98%~99%.

6. The use of any one of the peptide-containing skin care compositions with anti-wrinkle and firming effects as described in claims 1 to 5 in cosmetics.

7. The application of the peptide-containing skincare composition with anti-wrinkle and firming effects as described in claim 6 in cosmetics, characterized in that, It is added to cosmetics at a concentration of 0.1% to 1% by weight.

8. The application of the peptide-containing skincare composition with anti-wrinkle and firming effects as described in claim 6 in cosmetics, characterized in that, It is added to cosmetics at a concentration of 0.5% by weight.

9. The application of the peptide-containing skincare composition with anti-wrinkle and firming effects as described in claim 6 in cosmetics, characterized in that, The cosmetics include any one of the following: facial masks, lotions, serums, face creams, essence lotions, toners, lotions, moisturizing lotions, eye creams, facial cleansers, sunscreens, nourishing creams, massage creams, anti-wrinkle agents, acne treatments, after-sun preparations, and whitening agents.

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