Bioactive composition with anti-wrinkle effect and application

Abstract

The invention belongs to the technical field of biomedical cosmetology, and particularly relates to a bioactive composition with an anti-aging effect and application. The invention firstly provides a composition capable of promoting exogenous collagen to be integrated onto a collagen network. The composition is prepared from acetyl hexapeptide-8, snake venom-like peptide and PDRN. The composition can promote exogenous collagen to be integrated into an endogenous collagen network so as to achieve long-acting anti-aging and anti-wrinkle effects.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medical cosmetology, and particularly relates to a biological active composition with anti-aging effect and application. BACKGROUND

[0002] Skin aging is a natural physiological process, and its core manifestation is the degradation of the extracellular matrix (ECM) of the dermis, especially the structural disorder of the collagen fiber network and the loss of collagen, which further leads to typical signs of aging such as skin wrinkles, laxity, and reduced elasticity, affecting skin appearance and quality of life. Collagen, as the most abundant structural protein in the dermis, is essential for maintaining the youthful appearance of the skin. To address this challenge, supplementing exogenous collagen has become one of the widely used strategies in the current cosmetic and medical fields, such as topical application or direct injection, the basic concept of which is to improve the appearance of the skin by increasing the content of collagen in the skin. However, the existing technology has great limitations in using collagen to reduce wrinkles, and the fundamental reason is that exogenous collagen is difficult to integrate into the complex collagen network in the body. Due to the lack of such integration, the anti-wrinkle effect of exogenous collagen supplementation is often limited to superficial moisturizing and temporary filling, and cannot achieve the repair and fundamental remodeling of the skin structure. Therefore, the effect of simply supplementing exogenous collagen is limited in duration, and frequent multiple use or injection is needed to maintain the apparent improvement, which cannot alleviate the structural problems leading to skin aging.

[0003] Patent with publication number CN120714103A discloses a regenerative filling gel containing type III collagen, type XVII collagen, and PDRN microspheres; another patent with publication number CN111228134A discloses a cosmetic product rich in snake venom-like peptides, acetyl hexapeptide-8, and carnosine, which mainly functions to fade wrinkles and tighten and anti-aging. Although the above-mentioned technology can alleviate wrinkles by providing temporary filling or short-term muscle relaxation, the components therein fail to form a multi-component organic synergy to truly promote the reconstruction and optimization of the collagen network, and therefore cannot achieve long-term and stable anti-wrinkle effect.

[0004] In view of the above, it is necessary to propose a new technical solution to improve the existing technology. SUMMARY

[0005] The present application aims to provide a biological active composition with anti-aging effect and application, which partially solves or alleviates the above-mentioned deficiencies in the prior art. The present application specifically adopts the following technical solutions.

[0006] In one aspect, the present application provides the use of a synergistic composition with biological activity.

[0007] A composition for promoting integration of exogenous collagen into collagen network, the composition comprising acetyl hexapeptide-8, a snake venom-like peptide and PDRN. The exogenous collagen comprises type I collagen, type III collagen or a combination thereof.

[0008] In some preferred embodiments, the composition consists of acetyl hexapeptide-8, a snake venom-like peptide and PDRN.

[0009] The "collagen network" as used herein refers to a natural structure present in the structure of human skin, which is a core scaffold structure for maintaining the youthful and healthy function of skin.

[0010] The "integration" as used herein refers to a biological phenomenon, an active, regulated and structure-function integrated biological process, through the synergistic effect of specific multi-components, the exogenous collagen and endogenous collagen form a whole, that is, through the specific components, the exogenous collagen is combined with the endogenous collagen in a persistent structural and functional manner in the collagen network of the skin, so as to stably repair and reshape the collagen network structure of the skin, and achieve the purpose of long-acting anti-wrinkle.

[0011] Further, the composition comprises the following ingredients in the following weight percentage: 0.001%~1% PDRN, 0.001%~4% snake venom-like peptide and 0.001%~2% acetyl hexapeptide-8.

[0012] As a preferred embodiment, the composition comprises the following ingredients in the following weight percentage: 0.1% PDRN, 1% snake venom-like peptide and 0.5% acetyl hexapeptide-8.

[0013] In another aspect, the present application provides a use of a bioactive composition having anti-aging effect.

[0014] A use of a bioactive composition having anti-aging effect in the preparation of a long-acting anti-wrinkle product, the bioactive composition comprising acetyl hexapeptide-8, a snake venom-like peptide, PDRN (a synergistic composition having bioactivity) and collagen.

[0015] Further, the bioactive composition comprises the following ingredients in the following weight percentage: 0.001%~1% PDRN, 0.001%~4% snake venom-like peptide, 0.001%~2% acetyl hexapeptide-8 and 0.01%~2% collagen.

[0016] Further, the collagen comprises type I collagen, type III collagen or a combination thereof.

[0017] Further, the type of the long-acting anti-wrinkle product comprises a solution, a gel, an emulsion, a cream, a serum, a mask, a microneedle, a freeze-dried powder and / or an injection.

[0018] Further, the acetyl hexapeptide-8, the snake venom-like peptide and the PDRN are used to promote the collagen to integrate onto the collagen network of the skin.

[0019] Another aspect of the present application provides a composition with long-acting anti-wrinkle effect.

[0020] A bioactive composition with long-acting anti-wrinkle effect, the bioactive composition comprising acetyl hexapeptide-8, snake venom-like peptide, PDRN and collagen; the collagen comprising type I collagen, type III collagen or a combination thereof.

[0021] Further, the bioactive composition comprises the following ingredients in the following weight percentage: 0.001%~1% PDRN, 0.001%~4% snake venom-like peptide, 0.001%~2% acetyl hexapeptide-8 and 0.01%~2% collagen.

[0022] As a preferred, the composition comprises the following ingredients in the following weight percentage: 0.1% PDRN, 1% snake venom-like peptide, 0.5% acetyl hexapeptide-8 and 0.5% collagen. Beneficial technical effects: The present application proposes a new biological composition for long-acting anti-aging by promoting the collagen to integrate onto the collagen network of the skin (dermal layer), the anti-aging biological composition comprising acetyl hexapeptide-8, snake venom-like peptide, PDRN and exogenous collagen (type I, type III or a combination thereof). In this composition, acetyl hexapeptide-8, snake venom-like peptide, PDRN as synergistic components with biological activity, promote and achieve the biological process of exogenous collagen (type I, type III or a combination thereof) to integrate onto the collagen network of the skin.

[0023] The present application discloses the deep synergistic mechanism among acetyl hexapeptide-8, snake venom-like peptide and PDRN as synergistic components with biological activity. In this system, PDRN not only promotes endogenous collagen synthesis, but also mediates the stable integration of exogenous collagen and skin matrix as a "guide", solving the problem of low integration degree of traditional collagen supplementation. At the same time, the nerve-like polypeptide surpasses the traditional function, as a "microenvironment optimization agent" significantly reduces mechanical stress by soothing muscles, providing a "low interference" biological environment for collagen synthesis and integration, thus realizing the qualitative change from "component superposition" to "system synergy", achieving comprehensive, persistent and natural anti-wrinkle effect. Experiments found that the anti-wrinkle effect of the biological composition provided by the present application can be maintained for 2~5 times longer than that of exogenous collagen (type I, type III or a combination thereof) alone. BRIEF DESCRIPTION OF DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description. In all the drawings, similar elements or parts are generally identified by similar reference signs. In the drawings, each element or part is not necessarily drawn according to the actual proportion. Obviously, the drawings described below are some embodiments of the present application, and for those skilled in the art, other drawings can also be obtained without creative labor on the basis of these drawings.

[0025] Figure 1 The mechanism diagram of the synergistic effect of PDRN, acetyl hexapeptide-8, and snake venom-like peptide on collagen (I, III) in one of the embodiments of the present application; Figure 2 The skin effect of PDRN, acetyl hexapeptide-8, and snake venom-like peptide on collagen (I, III) in one of the embodiments of the present application in animal experiments; Figure 3 The long-acting anti-wrinkle effect of the components of each test group in one of the embodiments of the present application. DETAILED DESCRIPTION

[0026] In order to make the purpose, technical solutions and advantages of the embodiments of the present application more clear, the following will combine the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are some of the embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0027] In this document, "and / or" includes any and all combinations of one or more of the associated items.

[0028] In this document, "a plurality of" means two or more, that is, it includes two, three, four, five, etc.

[0029] In this specification, the term "about" typically means + / - 5% of the stated value, more typically + / - 4% of the stated value, more typically + / - 3% of the stated value, more typically + / - 2% of the stated value, even more typically + / - 1% of the stated value, even more typically + / - 0.5% of the stated value.

[0030] In this specification, certain embodiments can be disclosed in a range format. It should be understood that the description in range format is merely for convenience and brevity and that one of skill in the art would understand that the range format is applicable to each and every specification, example, and embodiment of the present application. For example, a range of "1 to 6" is intended to include the endpoints, "1" and "6", and all intervening values of the range, such as "3" and "4", as well as fractions of the range, such as "1.8", between the endpoints. The same applies to ranges including integers within the discrete range, such as "1 to 3." In addition, it should be understood that the description in range format also applies to any range with endpoints, including fractional endpoints, not explicitly recited, but instead known in the art to which the application pertains.

[0031] The present application provides a composite anti-wrinkle composition, the active ingredients and the preferred proportion (in percentage by weight) range as follows.

[0032] PDRN (Polydeoxyribonucleotide): PDRN is derived from salmon DNA, and is a bioactive substance with cell regeneration and tissue repair capabilities. It can activate fibroblast activity, promote the synthesis of endogenous collagen (especially types I and III) and elastin in the skin, to enhance the structural support and elasticity of the skin. In addition, PDRN can also improve skin microcirculation and has anti-inflammatory effects.

[0033] Collagen: Collagen has fine and soft fibers, and plays an important role in maintaining skin elasticity and smoothness. Exogenous supplementation of collagen can directly provide immediate and effective structural support for the skin, but the effect is difficult to sustain.

[0034] Acetyl Hexapeptide-8 (Argireline): Acetyl Hexapeptide-8 is a bioactive peptide that mimics the N-terminal structure of SNAP-25 protein, which can reduce the release of acetylcholine at the nerve terminal by competitively inhibiting the formation of SNARE complex. This mechanism of action can reversibly and gently relax facial expression muscles, effectively reducing the formation and deepening of dynamic wrinkles caused by muscle contraction.

[0035] Dipeptide Diaminobutyroyl Benzylamide Diacetate (Syn-Ake): Dipeptide Diaminobutyroyl Benzylamide Diacetate is a synthetic peptide that mimics the action of Waglerin-1, a snake venom toxin, which can reversibly block nicotinic acetylcholine receptors and further inhibit muscle cell contraction.

[0036] The above-mentioned complex anti-wrinkle composition can also optionally contain pharmaceutically or cosmetically acceptable carriers and adjuvants, such as the following.

[0037] Solvents: water for injection, purified water, physiological saline, etc., to ensure the purity and biocompatibility of the product.

[0038] Moisturizers: sodium hyaluronate, glycerol, butylene glycol, etc., to maintain skin hydration and enhance product efficacy.

[0039] pH adjusters: phosphate buffer, citric acid, sodium hydroxide, etc., to adjust and maintain the physiological pH of the composition.

[0040] Stabilizers: polysorbate, EDTA, etc., to maintain the stability of active ingredients and extend the shelf life of the product.

[0041] Preservatives: phenoxyethanol, parabens, etc. (if it is a non-injection product), to ensure the microbiological safety of the product.

[0042] Other active ingredients: niacinamide, vitamin C derivatives, antioxidants, etc. (if it is a non-injection product), to provide more comprehensive skin care efficacy.

[0043] Example 1 Mechanism and verification of the composition of the present application in synergistic collagen integration.

[0044] Experimental procedure: This example uses C57BL / 6 mice. A blank control group, a 0.5% collagen group, and a 0.1% PDRN + 1% snake venom peptide + 0.5% acetyl hexapeptide-8 + 0.5% collagen group (preferred composition of the present application) are set up. The drug is administered by subcutaneous or intradermal injection.

[0045] Sample processing and staining: the experimental animal models are sacrificed after 3 weeks of drug administration, and the skin in the drug administration area is removed. After fixation and paraffin embedding, 5 μm thick sections are prepared. The sections are stained with 0.1% Sirius red picric acid saturated solution for 60 minutes in the dark, and then differentiated with 0.01N hydrochloric acid and quickly dehydrated, transparent, and mounted.

[0046] Image acquisition and analysis: observe the sections under a microscope to evaluate collagen integration and structural remodeling. The section image is shown in Figure 2 .

[0047] Experimental results: Through Figure 2The histological section results clearly show that the composition group of the present application (containing 0.1% PDRN + 1% snake venom peptide, 0.5% acetyl hexapeptide-8 and 0.5% collagen group) has a significant advantage in collagen integration. Compared with the collagen group alone, the collagen network of the composition group shows a more precise, ordered and thick arrangement structure, and the section staining is deeper, indicating that the collagen content or density is significantly improved. Notably, the collagen network integration effect of the collagen group alone is slightly better than that of the blank group, but the improvement degree is far less than that of the composition group of the present application. This fully proves that PDRN, acetyl hexapeptide-8 and snake venom peptide can synergistically promote the effective integration of exogenous collagen and the remodeling of endogenous collagen network, thereby constructing a more dense, ordered and stable dermal collagen network. The mechanism diagram of the composition of the present application for promoting the integration of exogenous collagen and endogenous collagen is shown in Figure 1 .

[0048] Long-term anti-wrinkle effect verification of the composition of the present application. Construction of a wrinkle mouse model: Experimental animals: hairless mice (SKH-1 strain) Model construction: UVB lamps were used for groups other than the blank group, with an initial dose of 50 mJ / cm². Skin reactions were observed after each irradiation 3 times a week. The dose was gradually increased until the mouse back skin showed obvious erythema, roughness, decreased elasticity and wrinkle formation.

[0049] Experimental grouping and administration: After the successful establishment of the photoaging model, the mice in each group were given a one-time intradermal injection. Each mouse was injected with a total of 100 μL, which was evenly injected at 4-6 points on the depilated area of the back Grouping and administration: blank control group (no UVB irradiation, no drug administration). Model control group (UVB irradiation, injection of carrier).

[0050] Collagen group: 0.5% collagen PDRN + collagen group: 0.1% PDRN + 0.5% collagen.

[0051] Snake venom peptide + acetyl hexapeptide-8 + collagen group: 1% snake venom peptide + 0.5% acetyl hexapeptide-8 + 0.5% collagen.

[0052] Composition group of the present application (mixed and administered in the preferred ratio): 0.1% PDRN + 1% snake venom peptide + 0.5% acetyl hexapeptide-8 + 0.5% collagen group.

[0053] Evaluation index and method: Wrinkle improvement rate Methods: Skin imprints were collected from the depilated area of the back of the mice at pre-dose (Week 0) and post-dose at Week 1, Week 3, Week 5. The imprints were scanned using a high-resolution skin three-dimensional topography measuring instrument (e.g. PRIMOS or VISIOSCAN) to quantify the skin surface roughness parameters (e.g. Ra, Rz, Rt) and to calculate the wrinkle depth, length and number.

[0054] Histological Analysis: At the end of the experiment (Week 5), some mice were sacrificed and the dorsal skin tissue was fixed, embedded, sectioned and stained with Sirius red. The evaluation criteria are shown in Table 1 and Figure 3 .

[0055] Table 1 Skin wrinkle improvement rate In combination Figure 3 As can be seen from Table 1, the anti-wrinkle effect of collagen alone is limited after a single intradermal administration. Although the combination of PDRN with collagen, and the combination of acetyl hexapeptide-8 and snake venom-like peptide with collagen, can enhance the anti-wrinkle effect in the short term, these effects are not sustainable. However, when PDRN, snake venom-like peptide and acetyl hexapeptide-8 are combined with collagen (composition group of the present application), the anti-wrinkle effect is significantly enhanced and can be maintained at a high level for a long time. This excellent and long-lasting performance strongly supports the synergistic effect of PDRN, snake venom-like peptide and acetyl hexapeptide-8 with collagen, which can produce an effect far exceeding that of a single component or partial combination in terms of anti-wrinkle. This remarkable efficacy maintenance ability is attributed to the unique synergistic effect of the active ingredients in the formula, which promotes the in vivo integration of exogenous collagen and the endogenous repair and regeneration of the skin, thereby achieving significant synergistic effect and sustainability that is not simply additive.

[0056] Example 2 In vivo animal model experiments were conducted to verify the anti-wrinkle effect, collagen integration ability and safety of the composite anti-wrinkle composition.

[0057] Experimental animals: SKH-1 hairless mice.

[0058] Model construction: Ultraviolet (UVB) irradiation was used to induce skin photoaging, and an animal model with obvious wrinkles was successfully constructed.

[0059] Grouping and administration: blank control group (no UVB irradiation, no administration). Model control group (UVB irradiation, injection of carrier). Single active ingredient group (PDRN, collagen, acetyl hexapeptide-8, snake venom-like peptide, administered separately).

[0060] The present application is a composite anti-wrinkle composition group (PDRN + collagen + acetyl hexapeptide-8 + snake venom peptide).

[0061] Dosing method: Intradermal injection, lasting for 4 weeks.

[0062] Evaluation index and method: Before the start of treatment, at the middle of treatment and at the end of treatment, all animals are measured for the following indicators.

[0063] Preparation before measurement: The animals need to be lightly anesthetized before measurement to ensure stability during measurement.

[0064] Measurement indicators: Improvement rate of skin appearance: High-resolution image acquisition: Use professional skin image acquisition systems (such as Visioface, PRIMOS, etc.) or high-resolution digital cameras to take standardized photos of the animals' back skin under fixed angle and fixed lighting conditions.

[0065] Image analysis software: Use image analysis software to process images and quantify parameters such as wrinkle depth, length, number and roughness.

[0066] Wrinkle improvement rate = [(pre-treatment wrinkle parameter - post-treatment wrinkle parameter) / pre-treatment wrinkle parameter] ×100%.

[0067] Improvement rate of skin elasticity: Use Cutometer skin elasticity measurement instrument to measure the deformation and recovery ability of the skin by negative pressure suction and provide a series of elasticity parameters.

[0068] Measurement method: Select 3-5 fixed points on the depilated area of the animals' back for measurement, measure each point 3 times and take the average value. Ensure that the probe is perpendicular to the skin surface and maintain constant pressure.

[0069] Elasticity improvement rate = [(post-treatment elasticity parameter - pre-treatment elasticity parameter) / pre-treatment elasticity parameter] ×100%.

[0070] Histopathological analysis: Sampling: At the end of the experiment, skin biopsy samples are taken from the wrinkle area of the animal model.

[0071] Processing: Biopsy samples are fixed with 10% formalin, dehydrated, paraffin-embedded, and then cut into 5-micron-thick continuous sections.

[0072] Staining: Masson's Trichrome staining: Used to evaluate the density, arrangement and maturity of collagen fibers.

[0073] Verhoeff-Van Gieson (VVG) staining: used to evaluate the integrity, number and arrangement of elastic fibers.

[0074] Analysis: The stained sections were observed under an optical microscope and analyzed in combination with image analysis software. The evaluation indicators are shown in Tables 2 and 3.

[0075] Table 2 Effects of different ingredients on wrinkle improvement rate and skin elasticity improvement rate Table 3 Results of skin wrinkle and elasticity detection According to the data in Tables 2 and 3, the single ingredient (PDRN, collagen, echistatin-like peptide or acetyl hexapeptide-8) has limited improvement on wrinkles and elasticity (wrinkle improvement rate 8.9%-12.8%, elasticity improvement rate 7.1%-9.5%), and the histological score is only 2-3 points, which is difficult to achieve deep repair. The efficacy of the two-component combination (such as PDRN + collagen) is improved, the wrinkle improvement rate reaches 22.3% ± 3.8%, the elasticity improvement rate reaches 19.8% ± 3.5%, and the collagen fiber arrangement score reaches 4 points, showing a preliminary synergistic effect. The four-component combination of the present application (test examples 1-3) shows significant superiority, among which test example 2 (PDRN 0.1%, collagen 0.5%, echistatin-like peptide 1%, acetyl hexapeptide-8 0.5%) has the best effect, the wrinkle improvement rate reaches 48.1%, the elasticity improvement rate reaches 42.5%, and the histological score reaches 5 points, far exceeding the single and two-component combinations, confirming the strong synergistic effect. The data of the comparative examples show that the dosage range of each component is crucial. When the dosage is lower than the lower limit, the effect decreases significantly, and the score is only about 3 points; when the dosage is higher than the upper limit, the efficacy does not further improve, indicating that exceeding the range may lead to a plateau of effect or a decrease in efficiency, and even may affect the fine integration of fiber structure.

[0076] It should be noted that in this document, the terms "comprising", "including", or any other variant thereof are intended to cover non-exclusive inclusions, so that processes, methods, articles or devices that include a series of elements not only include those elements, but also include other elements not explicitly listed, or inherent to such processes, methods, articles or devices. Without more limitations, the element defined by the statement "including a" does not exclude the presence of another identical element in the process, method, article or device that includes the element.

[0077] The embodiments of the present application are described above with reference to the accompanying drawings, but the present application is not limited to the above-described specific embodiments, and the above-described specific embodiments are merely illustrative, but not restrictive, and a person of ordinary skill in the art can make many forms under the inspiration of the present application without departing from the purpose of the present application and the scope protected by the claims, and these all belong to the protection of the present application.

Claims

1. A composition for promoting integration of exogenous collagen into a collagen network, characterized in that, The composition consists of acetyl hexapeptide-8, a snake venom peptide and PDRN.

2. The composition of claim 1, wherein The composition includes the following ingredients in weight percent: 0.001-1% PDRN, 0.001-4% snake venom peptide and 0.001-2% acetyl hexapeptide-8.

3. The composition of claim 2, wherein The composition includes the following ingredients in weight percent: 0.1% PDRN, 1% snake venom peptide and 0.5% acetyl hexapeptide-8.

4. Use of a biologically active composition having an anti-aging effect for the preparation of a long-lasting anti-wrinkle product, characterized in that, The bioactive composition includes acetyl hexapeptide-8, a snake venom peptide, PDRN and collagen.

5. Use according to claim 4, characterized in that, The bioactive composition includes the following ingredients in weight percent: 0.001-1% PDRN, 0.001-4% snake venom peptide, 0.001-2% acetyl hexapeptide-8 and 0.01-2% collagen.

6. The use according to claim 4, wherein the compound is ###0002### The collagen includes type I collagen, type III collagen or a combination thereof.

7. The use according to claim 4, wherein the compound is ###0002### The long-lasting anti-wrinkle product is in the form of a solution, a gel, an emulsion, a cream, a serum, a mask, a microneedle, a lyophilized powder and / or an injection.

8. The use according to claim 4, wherein the compound is ###0002### The acetyl hexapeptide-8, the snake venom peptide and the PDRN are used to promote the integration of the collagen onto the endogenous collagen network.

9. A biologically active composition having long-lasting anti-wrinkle effects, characterized in that, The bioactive composition includes acetyl hexapeptide-8, a snake venom peptide, PDRN and collagen; the collagen includes type I collagen, type III collagen or a combination thereof.

10. The biologically active composition of claim 9, wherein The bioactive composition includes the following ingredients in weight percent: 0.001-1% PDRN, 0.001-4% snake venom peptide, 0.001-2% acetyl hexapeptide-8 and 0.01-2% collagen.

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