An active composition for promoting collagen regeneration and its preparation method and application

Abstract

The present invention discloses an active composition for promoting collagen regeneration, a preparation method and application thereof, and belongs to the field of cosmetics. The composition is composed of an extract of Staghornia ditchii, an extract of Pheretima calyx, deer bone collagen, tripeptide-1, and oligopeptide-2. The present invention has a synergistic effect by compounding the raw materials, thereby obtaining an excellent effect of promoting collagen regeneration.

Description

Technical Field

[0001] The present application relates to the field of cosmetics, and in particular to an active composition for promoting collagen regeneration, and a preparation method and application thereof. Background Art

[0002] Collagen is the main component of the extracellular matrix (ECM). It is mainly found in human skin, bones, cartilage, teeth, tendons, ligaments and blood vessels. It is an important structural protein of connective tissue. As the main component of the dermis of human skin, collagen and a small amount of elastin together form a regular collagen fiber network structure, which gives the skin a certain elasticity and hardness and transports moisture to the epidermis. However, factors such as aging, ultraviolet radiation, smoking and unhealthy lifestyles can cause the loss of human collagen. With the loss of a large amount of collagen, the skin will gradually show characteristics of skin aging such as decreased elasticity, loose wrinkles, pigmentation, dryness, roughness and sensitivity.

[0003] Therefore, it is necessary to develop an active composition that can promote collagen regeneration to improve the aging characteristics caused by the massive loss of collagen. Summary of the invention

[0004] The purpose of the present invention is to provide an active composition for promoting collagen regeneration, a preparation method and application thereof, which is prepared using plant extracts as raw materials, has high safety and has excellent effects on promoting collagen regeneration.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides an active composition for promoting collagen regeneration, comprising the following components: an extract of Acropora dactylifera, an extract of Pheretima pubescens, deer bone collagen, tripeptide-1, and oligopeptide-2.

[0007] Preferably, the active composition for promoting collagen regeneration consists of the following components in parts by weight:

[0008] Extract of Psoralea corylifolia: 4-6 parts; Extract of Pheretima sutchuenensis: 5-8 parts; Deer bone collagen: 5-7 parts;

[0009] Tripeptide-1: 1-3 parts; Oligopeptide-2: 1-3 parts.

[0010] Further preferably, the active composition for promoting collagen regeneration consists of the following components in parts by weight:

[0011] Extract of Pterocarpus dulse: 5-6 parts; Extract of Pheretima pubescens: 5-6 parts; Deer bone collagen: 5-6 parts; Tripeptide-1: 2-3 parts; Oligopeptide-2: 1-2 parts.

[0012] Most preferably, the active composition for promoting collagen regeneration consists of the following components in parts by weight:

[0013] Extract of Pterocarpus dulse: 5 parts; Extract of Pheretima pubescens: 6 parts; Deer bone collagen: 6 parts; Tripeptide-1: 2 parts; Oligopeptide-2: 2 parts.

[0014] In a second aspect, the present invention provides a skin care product, wherein the skin care product is composed of the following components in percentage by weight:

[0015] 1-3wt% of the active composition for promoting collagen regeneration described in the first aspect;

[0016] Glycerol 20-30wt%;

[0017] Sodium ascorbate 1-5wt%;

[0018] Deionized water balance.

[0019] In a third aspect, the present invention provides a method for preparing the skin care product according to the second aspect, the specific steps are as follows:

[0020] The active composition for promoting collagen regeneration is mixed with deionized water and homogenized, glycerol and sodium ascorbate are added, stirred and mixed, homogenized, and discharged to obtain a skin care product, which is sealed and stored in a dark place at 2-4°C.

[0021] The skin care product dosage form can be any one of toner, essence, lotion, mask, gel and cream.

[0022] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention provides an active composition for promoting collagen regeneration, a preparation method and an application thereof, wherein the composition having the effect of promoting collagen regeneration is compounded by a mixture of a ditch stag beetle extract, a calyx earthworm extract, deer bone collagen, tripeptide-1 and oligopeptide-2. The ditch stag beetle extract contains abundant natural active substances such as carrageenan, D-alginic acid, and plant hormone precursors; the calyx earthworm extract contains abundant collagenase, plasmin and other proteases with anticoagulant and fibrinolytic effects, and has the effect of indirectly participating in collagen metabolism and repair; deer bone collagen is rich in elastin, and has a certain effect of improving skin sagging and delaying aging; oligopeptide-2, tripeptide-1 and other polypeptide components have a significant effect of promoting collagen production. The research results show that the combined use of ditch stag beetle extract, calyx earthworm extract, deer bone collagen, tripeptide-1 and oligopeptide-2 has a significant synergistic effect and can significantly stimulate the production of collagen. DETAILED DESCRIPTION

[0023] In order to better understand the present invention, the present invention is further described below in conjunction with specific application examples, wherein the terms used in the application examples are for describing specific implementation schemes and do not constitute limitations on the protection scope of the present invention.

[0024] The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the calculation is based on mass percentage and mass fraction.

[0025] Some raw materials and their sources are as follows:

[0026] Psoralea corylifolia extract: purchased from Xiamen Xulijin Biotechnology Co., Ltd.

[0027] Pheretima earthworm extract: purchased from Xi'an Changyue Biotechnology Co., Ltd.

[0028] Deer bone collagen: purchased from Shaanxi Zelang Biotechnology Co., Ltd.

[0029] Tripeptide-1: purchased from Shenzhen Vicky Technology Co., Ltd.

[0030] Oligopeptide-2: purchased from Shenzhen Weiqi Technology Co., Ltd.

[0031] The mass proportion of the raw materials of the composition for promoting collagen regeneration is as follows:

[0032] Table 1 Mass ratio of the composition for promoting collagen regeneration

[0033]

[0034] Note: The missing components in composition ①-⑦ are replaced with deionized water of equal mass.

[0035] The skin care product described in the present application is specifically an essence liquid that promotes collagen regeneration.

[0036] The mass percentage of raw materials for promoting collagen regeneration essence is as follows:

[0037] Components Application Example 1 Application Example 2 Application Example 3 Application Example 4 Application Example 5 Blank Application Example Composition for promoting collagen regeneration / wt% 2 3 1 2 2 0 Composition No. Composition 1 Composition 2 Composition 3 Composition 1 Composition 1 - Glycerol / wt% 20 25 30 20 20 20 Sodium ascorbate / wt% 4 1 5 4 4 4 Deionized water Replenish to 100% Replenish to 100% Replenish to 100% Replenish to 100% Replenish to 100% Replenish to 100%

[0038] The preparation methods of the above application examples 1-5 and blank application examples are as follows:

[0039] At room temperature, the active composition for promoting collagen regeneration and deionized water are placed in an emulsifying pot and mixed and homogenized; glycerol and sodium ascorbate are added while stirring, and stirring is continued for 20-25 minutes after the glycerol and sodium ascorbate are added; homogenization is performed, and the material is discharged to obtain a finished essence liquid; the essence liquid is sealed and packaged, and stored at 2-4°C away from light.

[0040] Experiment 1: Experiment on promoting collagen regeneration

[0041] Refer to the Chinese patent authorization number: CN114540457B "A test method for the anti-wrinkle effect of cosmetics and their raw materials" and modify it. The specific steps are as follows:

[0042] Samples: Compositions 1-3, compositions ①-⑦.

[0043] Cells: Human skin fibroblasts (HSF cells), purchased from: STM-CL-5176.

[0044] Kit: Human type I collagen (ColⅠ) ELISA kit, purchased from: Shanghai ELISA Biotechnology Co., Ltd. Model: mL057630.

[0045] In the experiment, the reagents and consumables required for cell culture were purchased from Thermo Fisher Scientific Gibco.

[0046] The frozen human skin fibroblasts were taken out from the liquid nitrogen tank for cell recovery and subculture.

[0047] The specific method of recovery culture is: quickly dissolve the frozen cells in a 37°C water bath, centrifuge at 800rpm for 8 minutes, discard the supernatant, resuspend the bottom cells with DMEM culture medium containing 10% bovine serum, transfer them to a T25 cell bottle, and culture them at 37°C and 5.0% CO2 for 36 hours. When the cell fusion degree reaches more than 80%, subculture is carried out.

[0048] The specific method of subculture is as follows: discard the cell culture medium, gently wash once with fresh serum-free culture medium or PBS, discard, add an appropriate amount (to immerse all the cell monolayer surface) of 0.25% EDTA-trypsin solution to digest for 4 minutes, discard the EDTA-trypsin solution, add 10mL of DMEM culture medium containing 10% bovine serum, and disperse the cells by blowing to disperse them evenly. Take the cell suspension and count it, and adjust the cell concentration to 3.0×10 according to the cell number. 4 ~5.0×10 4 cell / mL and culture at 37°C and 5.0% CO2 for 24 to 72 hours.

[0049] To ensure the viability and stability of the cells, this method was tested using cells of more than 2 generations after recovery, and the number of cell generations after recovery was controlled within 30 generations.

[0050] Adjust the cell concentration to 4.0*10 4 cell / mL, add to 96-well cell culture plate, 100 μL / well. Cover the cell plate, seal the edges, and culture at 37°C and 5.0% CO2 for 24 hours for sample collagen expression test;

[0051] The culture medium was discarded from the cultured cell plate, and the cell plate was washed 1-2 times with PBS. The dose of the cell plate was 500 mJ / cm 2 UVB was used to induce modeling. After irradiation, the sample was diluted to 2wt% with DMEM and added to the cell plate. Six wells were added to each well at the same concentration. A blank control group and a positive control group were set up at the same time. The blank control group was added with 100μL / well DMEM, and the positive control was added with 100μL / well DMEM medium containing 100ng / mL recombinant human transforming growth factor TGF-β. Six wells were added to each group. After the addition of samples, the cells were cultured at 37°C and 5.0% CO2 for 24 hours. After the culture was completed, the cell culture medium was discarded, and the cells were gently rinsed twice with PBS and discarded. The cells at the bottom were frozen and thawed or lysed, and the frozen and thawed liquid or lysed liquid was collected to determine the collagen content. During the test, two wells were randomly selected from each group of 6 wells and mixed into three samples for testing. The cell freezing and thawing method is: add 100μL / well PBS, place in a refrigerator below -15℃ and freeze and thaw twice repeatedly, collect the frozen and thawed solution for use; the cell lysis method is: add 100μL / well cell lysis solution, place in a 2-8℃ refrigerator for lysis for 10 minutes, collect the lysis solution for use. If it is not possible to measure in time, the cell fluid can be collected in a 1.5mL sterile centrifuge tube and placed in a -80℃ ultra-low temperature refrigerator for storage.

[0052] The collagen content of the cell culture fluid was tested by enzyme-linked immunosorbent assay (ELISA), and the specific test method was carried out according to the instruction manual of the human collagen enzyme-linked immunosorbent assay kit. According to the standard curve of the ELISA kit, the collagen content of the sample group, blank control group and positive control group was calculated respectively.

[0053] Collagen expression up-regulation rate (%) = (TC) / C × 100%;

[0054] Wherein, T refers to the average collagen content of the sample group, and C refers to the average collagen content of the blank control group.

[0055] Table 2 Test for promoting collagen regeneration

[0056] Composition Collagen upregulation rate / % Significance Composition 1 17.43 * Composition 2 17.51 * Composition 3 16.72 * Composition ① 10.36 * Composition ② 10.72 * Composition ③ 11.24 * Composition ④ 9.94 * Composition ⑤ 9.63 * Composition ⑥ 9.82 * Composition ⑦ 9.41 * blank - - Positive 104.76 **

[0057] Note: “*” indicates p<0.05

[0058] According to the experimental results in Table 2, the composition provided by the present invention has a significant effect of promoting collagen regeneration. According to the test results of composition 1 and compositions ①-⑦, there is a certain synergistic effect between the extract of Acropora dactylifera, the extract of Pheretima pubescens, deer bone collagen, tripeptide-1 and oligopeptide-2 in the composition.

[0059] Test 2: Patch test:

[0060] The present invention experiments verify the biosafety of application examples 1-5 and blank application examples.

[0061] The 2015 "Technical Specifications for Safety of Cosmetics" was used as a reference standard for the irritation evaluation of cosmetics. The test method was a skin patch test, and 60 people aged 16-65 years were randomly distributed and tested.

[0062] Test method: Put the test substance into the patch tester, the dosage is 0.020-0.025g, cover the patch tester with the test substance on the back or forearm of the subject with non-irritating cloth-based tape, and gently press it with the palm to make it evenly adhere to the skin surface for 24 hours. After removing the test patch tester, observe the skin reaction after the indentation disappears 30 minutes later. If the result is negative, observe again 24 hours and 48 hours after the patch test.

[0063] Evaluation criteria:

[0064] Grade 0: negative reaction;

[0065] Grade 1: Suspicious reaction, only slight erythema;

[0066] Grade 2: weak positive reaction, erythema, infiltration, edema, and papules may be present;

[0067] Grade 3: Strong positive reaction, erythema, infiltration, edema, papules, and the reaction may extend beyond the test area;

[0068] Grade 4: Very strong positive reaction, obvious erythema, severe infiltration, edema, confluent herpes, and the reaction exceeds the test area.

[0069] Test results: Skin reactions were negative in all subjects.

[0070] The embodiments described above are part of the embodiments of the present application, rather than all of the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents the selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present application.

Claims

1. A serum for promoting collagen regeneration, characterized in that: Contains the following components by mass percentage: Composition for promoting collagen regeneration: 1-3wt%; Glycerol: 20-30wt%; Sodium ascorbate: 1-5wt%: Deionized water to 100wt%: The composition for promoting collagen regeneration is composed of the following components in parts by weight: Carrageenan Extract: 4-6 parts; Pheretima pubescens extract: 5-8 parts; Deer bone collagen: 5-7 parts; Tripeptide-1: 1-3 parts; Oligopeptide-2: 1-3 parts.

2. A method for preparing the essence according to claim 1, characterized in that: The preparation method is specifically as follows: At room temperature, the collagen regeneration promoting composition and deionized water are placed in an emulsifying pot and mixed and homogenized; glycerol and sodium ascorbate are added while stirring, and stirring is continued for 20-25 minutes after the glycerol and sodium ascorbate are added; homogenization is performed, and the material is discharged to obtain a finished essence liquid; the essence liquid is sealed and packaged, and stored at 2-4°C away from light.

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