CN117679344B - Anti-aging repairing composition and preparation method and application thereof - Google Patents
Anti-aging repairing composition and preparation method and application thereof Download PDFInfo
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Abstract
The application discloses an anti-aging repairing composition, a preparation method and application thereof, wherein the anti-aging repairing composition comprises the following components in percentage by mass: 0.2-4% of pea extract, 0.01-1% of recombinant XVII type collagen, 1-20% of hydroxypropyl tetrahydropyran triol and the balance of solvent. The components of the composition act on the epidermis layer, the basement membrane and the dermis layer of the skin respectively, and the expression of related genes COL7, COL17, laminin5, integrin ITG-beta 4 and nestin Nidogen are promoted to promote the synergistic effect of multiple ways such as hemidesmosomes on the basal cells of the epidermis so as to improve the signal transmission and the substance transmission between dermis and epidermis, fully display the tension of the skin, improve the skin wrinkles, and simultaneously repair the skin barrier and improve the anti-aging repair effect.
Description
Technical Field
The application relates to the technical field of daily chemical products, in particular to an anti-aging repairing composition, a preparation method and application thereof.
Background
The basement membrane-semidesmosome can organically combine the epidermis layer and the dermis layer, strengthen the link between the epidermis layer and the dermis layer, and facilitate the signal transmission and the substance exchange of the skin from inside to outside, thereby achieving the effects of resisting wrinkles, smoothing the skin and recovering the water and light muscles. Wherein, the basement membrane is an important bearing structure between the epidermis and the dermis, and plays a role of double faced adhesive tape; the hemidesmosome is a main connection structure between epidermal basal cells and a basal membrane band below, is formed by embedding irregular protrusions of a cell membrane at the dermis side of keratinocytes and the basal membrane band, and is a key structure that the epidermis can adhere to the basal membrane. However, with age, the number of hemidesmosomes decreases, the adhesion of the basal lamina to the epidermis layer decreases, resulting in a slow down of the exchange of substances between the epidermis layer and the dermis layer, increasing skin wrinkles and exacerbating skin collapse.
At present, no anti-aging product which organically combines anti-aging and repairing aiming at a basement membrane-semi-desmosome to strengthen the anti-aging effect of skin exists in the market, and the anti-aging can not be comprehensively carried out from multiple layers of epidermis layers, basement membranes, dermis layers and the like, so that the epidermis and dermis of the skin can not be truly and organically communicated, and the overall anti-aging and repairing effects can be realized.
Disclosure of Invention
The application provides an anti-aging repairing composition, a preparation method and application thereof, and aims to solve the technical problem that the current anti-aging product cannot realize anti-aging from multiple layers such as a epidermis layer, a basal layer and a dermis layer.
The application provides an anti-aging repairing composition which comprises the following components in percentage by mass:
0.2-4% of pea extract, 0.01-1% of recombinant XVII type collagen, 1-20% of hydroxypropyl tetrahydropyran triol and the balance of solvent.
In one embodiment, the composition comprises the following components in percentage by mass:
pea extract 0.5-2%, recombinant XVII type collagen 0.01-0.5%, hydroxypropyl tetrahydropyran triol 5-10%, and the rest is solvent.
In one embodiment, the composition comprises the following components in percentage by mass:
pea extract 1.5%, recombinant XVII type collagen 0.2%, hydroxypropyl tetrahydropyran triol 8%, and the rest is solvent.
In one embodiment, the solvent comprises deionized water and an alcoholic solvent comprising one or more of propylene glycol, 1, 3-propanediol, butylene glycol, dipropylene glycol, glycerin, 1, 2-hexanediol, and 1, 2-pentanediol.
The components of the anti-aging repair composition act on the epidermis layer, the basement membrane and the dermis layer respectively, and the expression of related genes COL7, COL17, laminin Lamine 5, integrin ITG-beta 4 and nestin Nidogen are promoted to promote the synergistic effect of multiple ways such as hemidesmosomes on epidermal basal cells so as to improve the signal transmission and the substance transmission between dermis and epidermis, fully display the tension of skin, improve skin wrinkles and repair skin barriers at the same time.
Wherein the pea extract is rich in plant polypeptides with protease resisting activity, and can promote synthesis of monisin, proteoglycan, type I/III/V collagen and elastin.
Recombinant human XVII type collagen is secreted and expressed by Pichia pastoris with high efficiency, has an amino acid sequence with more than 80% homology with human XVII type collagen, and can promote the expression of related genes COL7, COL17, laminin5, integrin ITG-beta 4 and nestin Nidogen.
Hydroxypropyl tetrahydropyran triol activates the synthesis of mucopolysaccharide, promoting the synthesis of type IV collagen and type VII collagen.
The application provides a preparation method of an anti-aging repair composition, which comprises the following steps:
Adding deionized water into a stirring pot, heating to 70-90 ℃ under stirring, preserving heat for 20-40 min, and cooling to 25-50 ℃;
Step two, adding pea extract, recombinant XVII type collagen, hydroxypropyl tetrahydropyran triol and an alcohol solvent into a stirring pot, and stirring and dissolving until no particles exist;
Transferring the materials in the stirring pot into an ultrasonic equipment pot, carrying out ultrasonic treatment for 5-30 min, filtering and discharging by 400 meshes, wherein the ultrasonic frequency is 18-22 kHz, and obtaining the anti-aging repairing composition.
In one embodiment, the first step includes:
Adding deionized water into a stirring pot, heating to 80 ℃ under stirring, preserving heat for 30min, and cooling to 40 ℃.
In one embodiment, the third step includes:
transferring the materials in the stirring pot into an ultrasonic equipment pot, performing ultrasonic treatment for 20min, and filtering and discharging at an ultrasonic frequency of 20kHz and 400 meshes to obtain the anti-aging repairing composition.
The preparation method of the anti-aging repair composition can improve the solubility of pea extract and enhance the hydrolysis resistance of recombinant XVII type collagen, thereby enhancing the activity retention time of the anti-aging repair composition in water.
The application of the anti-aging repairing composition in skin care products is characterized in that the skin care products comprise face cream, emulsion, facial mask or essence.
In one embodiment, the anti-aging repair composition is present in the skin care product at a percentage of 3%
The skin care product contains an anti-aging repairing composition, can play a synergistic effect aiming at most of key targets of basement membrane and semidesmosomes, strengthen signal transmission and substance exchange between dermis and epidermis, achieve the effects of more rapidly, continuously and effectively resisting wrinkles, repairing skin and reducing skin darkness and roughness.
Drawings
FIG. 1 is a forehead line image of subject 1 before and after 28 days of use of an anti-aging facial cream, as shown in an embodiment of the present application;
FIG. 2 is a forehead line image of subject 2 before and after 28 days of anti-aging facial cream use, as shown in an embodiment of the application;
Fig. 3 is a forehead line image of subject 3 before and after 28 days of using an anti-aging facial cream, as shown in an embodiment of the present application.
Detailed Description
The following detailed description of the present invention will provide further details in order to make the above-mentioned objects, features and advantages of the present invention more comprehensible. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. The present invention may be embodied in many other forms than described herein and similarly modified by those skilled in the art without departing from the spirit of the invention, whereby the invention is not limited to the specific embodiments disclosed below.
The term "prepared from …" as used herein is synonymous with "comprising". The terms "comprising," "including," "having," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, step, method, article, or apparatus.
When an equivalent, concentration, or other value or parameter is expressed as a range, preferred range, or a range bounded by a list of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when ranges of "1 to 5" are disclosed, the described ranges should be construed to include ranges of "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a numerical range is described herein, unless otherwise indicated, the range is intended to include its endpoints and all integers and fractions within the range.
Furthermore, the indefinite articles "a" and "an" preceding an element or component of the invention are not limited to the requirements of the number of elements or components (i.e. the number of occurrences). Thus, the use of "a" or "an" should be interpreted as including one or at least one, and the singular reference of an element or component also includes the plural reference unless the amount is obvious to the singular reference.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
The component raw materials and percentage contents of examples 1 to 3 and comparative examples 1 to 3 are shown in table 1:
TABLE 1 component percentage content table
The preparation methods of the anti-aging repair compositions of examples 1 to 3 and comparative examples 1 to 3 each include the following steps:
adding deionized water into a stirring pot, heating to 80 ℃ under stirring, preserving heat for 30min, and cooling to 40 ℃;
Step two, adding pea extract, recombinant XVII type collagen, hydroxypropyl tetrahydropyran triol and an alcohol solvent into a stirring pot, and stirring and dissolving until no particles exist;
Thirdly, transferring the materials in the stirring pot into an ultrasonic equipment pot, carrying out ultrasonic treatment for 20min, and filtering and discharging at an ultrasonic frequency of 20kHz and 400 meshes to obtain the anti-aging repairing composition.
The anti-aging repair compositions prepared in examples 1 to 3 and comparative examples 1 to 3 were subjected to in vitro tests, and the test methods and test results are as follows:
in vitro cell experiments: the method is characterized in that human immortalized skin keratinocyte HaCat cultured in vitro is taken as a model, the expression change of related proteins at the junction of the dermis after treatment by an anti-aging composition is tested, and the influence of the treatment by the composition on the expression of related functional genes of the basal membrane of the HaCat cell is detected by a qRT-PCR method.
QRT-PCR method, the test method is as follows:
First, extraction and reverse transcription of mRNA:
(1) Cells were plated in 6-well plates 1.5X105 cells per well, and after 24h 0.1% of the anti-senescence repair composition of example 1 was added for co-cultivation for 24h;
(2) Washing with PBS for 1 time, adding 1mL of Trizol, and transferring to a 1.5mL centrifuge tube or placing at-80 ℃ for later use;
(3) Centrifuging at 12000rpm for 5min at 4deg.C, removing precipitate, and collecting supernatant;
(4) Adding a certain amount of chloroform according to the proportion of 200 mu L of chloroform/1 ml of Trizol, shaking and mixing uniformly, and standing at room temperature for 15min;
(5) Centrifuging at 12000rpm for 15min at 4deg.C, sucking the supernatant into another clean centrifuge tube, carefully sucking, and if not, re-centrifuging to suck the supernatant;
(7) Adding a certain amount of isopropanol according to the ratio of 500 μl isopropanol/1 mLTrizol, mixing uniformly, and standing at room temperature for 10min;
(8) Centrifuging at 12000rpm for 10min at 4deg.C, sucking and removing supernatant to precipitate RNA at the bottom of the tube;
(9) Adding a certain amount of 75% ethanol according to the proportion of 1mL of 75% ethanol/1 mL of Trizol, and gently mixing;
(10) Centrifuging at 7500rpm for 5min at 4 ℃, sucking and removing supernatant, and standing for 5-10 min to dry RNA, but avoiding insufficient drying and excessive drying;
(11) Dissolving the precipitate with high-pressure DEPC water, gently blowing to dissolve completely, and placing on ice or storing in-80deg.C refrigerator;
(12) The spectrophotometer detects the OD260/280 value and quantifies the RNA concentration. Reverse transcription of RNA into cDNA using a reverse transcription kit is performed as follows:
① Genomic DNA removal reaction
The reaction mixture was prepared on ice using a reverse transcription (PRIMESCRIPT' MRTREAGENTKITWITHGDNAERASER) kit according to the following components, and in order to ensure the accuracy of the preparation of the reaction mixture, when each reaction was performed, masterMix was prepared in an amount of +2 of the reaction, and then was packed into each reaction tube, and finally RNA samples were added, and the reaction system was as shown in table 2.
TABLE 2 genomic DNA removal reaction System Table
② Reverse transcription reaction
Preparing reaction liquid on ice, preparing a MasterMix according to the reaction number of +2 when each reaction is carried out in order to ensure the preparation accuracy of the reaction liquid, and then sub-packaging 10 mu L of the MasterMix into each reaction tube, and gently mixing the mixture, wherein a reaction system is shown in a table 3.
TABLE 3 Table of reverse transcription reaction systems
③ Real-time fluorescence quantitative PCR (qRT-PCR)
A20. Mu.L real-time fluorescent quantitative PCR reaction system was prepared as shown in Table 4.
TABLE 4 real-time fluorescent quantitative PCR reaction System Table
Setting 3 compound holes for each sample, taking GAPDH gene as an internal reference, obtaining threshold cycle numbers (Ct) of the GAPDH and target genes, and calculating the relative expression quantity of the target genes by using the detected Ct values: 2- Δct (target gene) -CT (GAPDH) ].
Examples 2 and 3 and comparative examples 1 to 3 were subjected to in vitro tests in the same manner as described above, and test results were obtained as shown in table 5.
Table 5 table of results of in vitro tests of examples 1 to 3 and comparative examples 1 to 3
As can be seen from Table 5 above, the relative expression levels of the anti-aging treatment compositions of examples 1 to 3 were better for COL7, COL17, ITG-. Beta.4, laimin 5 and Nidogin genes than for comparative examples 1 to 3; obviously, compared with the comparative examples 1 to 3, the effect of promoting the expression of the basement membrane related genes is not obvious when pea extracts, recombinant XVII type collagen and hydroxypropyl tetrahydropyran triol are used alone, and the effect of promoting the expression of the basement membrane related genes is obvious when the components of examples 1 to 3 are compounded. It can be seen that the formulation components of examples 1 to 3 have a synergistic effect on gene expression of key structural components of the basement membrane and the hemidesmosomes.
Example 4
The anti-aging cream prepared from the anti-aging repairing composition is prepared from the anti-aging repairing composition prepared in the example 2, an oil phase component, an aqueous phase component and related auxiliary agents, and the percentage contents of the specific raw materials are shown in the table 6.
Table 6 percentage of cream ingredients
The preparation method of the anti-aging face cream comprises the following steps:
(1) Homogenizing the phase A component at 25 ℃ for 3 minutes until no particles are uniform, and heating to 85 ℃ to obtain a first mixed solution;
(2) Heating the phase B component to 85 ℃, and uniformly mixing to obtain a second mixed solution;
(3) Adding the second mixed solution obtained in the step (2) into the first mixed solution obtained in the step (1), and uniformly mixing to obtain a third mixed solution;
(4) Cooling the third mixed solution obtained in the step (3) to 50 ℃, adding the phase C, and uniformly mixing to obtain a fourth mixed solution;
(5) And (3) cooling the fourth mixed solution obtained in the step (4) to below 38 ℃, adding the phase D and the phase E, and uniformly mixing to obtain the anti-aging facial cream.
To demonstrate the effect and performance of the product of the invention, a product test was performed on the cream of example 4.
Test purpose: evaluating the efficacy of the test sample in anti-aging repair by normal continuous use of the test sample by adult female volunteers;
volunteer age: 40-60 years old;
Gender: female;
Volunteer population: 32 people;
volunteer facial skin texture: the facial skin is drier and rough, the facial skin is looser, the eyes have fine lines, the corners of the eyes have fine lines or wrinkles, the forehead has fine lines or wrinkles with obvious stature lines.
Product frequency of use: after the face is cleaned and the skin is refreshed, a proper amount of the product is taken, the product is smeared on the face and is gently rubbed and massaged until the product is absorbed, the product is used for 28 days, and the product is used for 2 times a day and each time in the morning and evening.
The evaluation method comprises the following steps: the subjects were tested for facial gloss, skin firmness F4, wrinkle depth (forehead lines, stature lines) before (D0) and after 28 days of use of the test product (D28), subject self-assessed questionnaires were conducted after 28 days of use of the test product (D28), and the test results before and after use of the product were compared by a statistical test method to determine whether there was a statistical difference.
The statistical method comprises the following steps: the statistical analysis software is SPSS. The metering values are normally distributed, and statistical analysis is carried out by adopting a t-test method; the metering values are non-normal distribution, and statistical analysis is carried out by adopting a rank sum test method. The statistical methods all used a two-tailed test, level=0.05.
The calculation formula is as follows:
improvement rate after 28 days of application of the product (skin firmness F4, wrinkle depth) = (D0-D28)/d0×100%;
Improvement rate after 28 days (gloss) with the product = - (D0-D28)/d0×100%;
Where D0 represents the skin parameter base value before the product is applied to the test area and D28 represents the skin parameter value after 28 days of the product application to the test area.
The test procedure for the subjects is shown in table 7.
Table 7 test flow schedule
(1) Test results
TABLE 8 skin gloss index Specification Table
Table 9 skin gloss analysis results table
1) The statistical method comprises the following steps: analysis was performed using t-test or rank sum test methods, with a test level α=0.05.
2) The significance labeling method comprises the following steps: "ns" means that there is no statistical difference, p.gtoreq.0.05; p <0.05 indicates a significant difference ("" indicates 0.01. Ltoreq.p <0.05 ";" indicates 0.001. Ltoreq.p <0.01 ";" indicates p < 0.001).
3) Number of product used = 32.
TABLE 10 skin firmness F4 specification table
Table 11 table of skin firmness F4 (mm x s) analysis results
1) The statistical method comprises the following steps: analysis was performed using a rank sum test method, with a test level of a=0.05.
2) The significance labeling method comprises the following steps: "ns" means no statistical difference, p20.05; p <0.05 indicates a significant difference ("" indicates 0.01. Ltoreq.p <0.05 ";" indicates 0.001. Ltoreq.p <0.01 ";" indicates p < 0.001).
3) Number of product used = 32.
Table 12 forehead line depth index specification table
TABLE 13 forehead line depth (. Mu.m) analysis results table
1) The statistical method comprises the following steps: analysis was performed using a t-test method, with a test level of a=0.05,
The significance labeling method comprises the following steps: "ns" means that there is no statistical difference, p.gtoreq.0.05; p <0.05 is indicated by a significant difference ("" means 0.01. Ltoreq.p <0.05 ";" means 0.001. Ltoreq.p <0.01 ";" means p < 0.001).
3) Number of product used = 32.
Wherein figures 1 to 3 show images of forehead lines of randomly selected 3 subjects, it can be seen from the figures that the forehead lines of the subjects are significantly shallower after the use of the anti-ageing face cream.
Table 14 subjects identify the product after 28 days of continuous use of the anti-aging cream
1) Evaluation index: the "very disagreement" is classified as 1,2, 3, 4, 5, and "disagreement".
2) Number of product used = 32.
As can be seen from tables 9, 11 and 13, the gloss of the subject was significantly reduced (p < 0.001) from the base value after 28 days of continuous use of the test example 4 product for 28 days, with an improvement of 40.99%; subject's skin firmness F4 was significantly reduced (p < 0.001) compared to basal value after 28 days of product use with an improvement rate of 38.77%; the subject's forehead line depth was significantly reduced (p < 0.001) from the basal value after 28 days of product use with an improvement rate of 32.80%.
From Table 14, it can be seen that 100.00% of subjects feel that the product is very moist after use; 96.88% of subjects feel that the product is more firm and has a pull-up feel after use; 100.00% of subjects feel that the product is finer and smoother in skin after use; 93.75% of the subjects felt the product to be full and elastic after use; 100.00% of subjects felt the product to be more transparent to the skin after use; 96.88% of subjects feel the product has moisturizing effect; 100.00% of subjects felt the product to have a soothing effect; 93.75% of subjects felt that the product could improve fish tail; 93.75% of subjects felt that the product could improve under-eye fine wrinkles; 93.75% of subjects felt that the product could improve forehead lines; 93.75% of the subjects felt the product to improve the french; 93.75% of subjects felt the product to improve the facial texture; 100.00% of subjects considered the product to be mild and non-irritating; 100.00% of the subjects were satisfied with the overall product.
Thus, it is possible to obtain the results of the tests in tables 9, 11, 13 and 14, that the anti-aging repairing composition of the present invention is applied to skin care products, and can repair skin while effectively reducing wrinkles, thereby achieving effects of reducing skin dullness and roughness.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The foregoing embodiments have been provided for the purpose of illustrating the general principles of the present application, and are not to be construed as limiting the scope of the application. It should be noted that any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art without departing from the spirit and principles of the present application are intended to be included in the scope of the present application.
Claims (9)
1. An anti-aging repairing composition is characterized by comprising the following components in percentage by mass:
0.2-4% of pea extract, 0.01-1% of recombinant XVII type collagen, 1-20% of hydroxypropyl tetrahydropyran triol and the balance of solvent.
2. An anti-aging repair composition according to claim 1, comprising the following components in mass percent:
pea extract 0.5-2%, recombinant XVII type collagen 0.01-0.5%, hydroxypropyl tetrahydropyran triol 5-10%, and the rest is solvent.
3. An anti-aging repair composition according to claim 2, comprising the following components in mass percent:
pea extract 1.5%, recombinant XVII type collagen 0.2%, hydroxypropyl tetrahydropyran triol 8%, and the rest is solvent.
4. An anti-aging treatment composition according to any one of claims 1 to 3, wherein the solvent comprises deionized water and an alcoholic solvent comprising one or more of propylene glycol, butylene glycol, dipropylene glycol, glycerol, 1, 2-hexanediol and 1, 2-pentanediol.
5. A method of preparing an anti-aging repair composition according to any one of claims 1 to 4, comprising the steps of:
Adding deionized water into a stirring pot, heating to 70-90 ℃ under stirring, preserving heat for 20-40 min, and cooling to 25-50 ℃;
Step two, adding pea extract, recombinant XVII type collagen, hydroxypropyl tetrahydropyran triol and an alcohol solvent into a stirring pot, and stirring and dissolving until no particles exist;
Transferring the materials in the stirring pot into an ultrasonic equipment pot, carrying out ultrasonic treatment for 5-30 min, filtering and discharging by 400 meshes, wherein the ultrasonic frequency is 18-22 kHz, and obtaining the anti-aging repairing composition.
6. A method of preparing an anti-aging repair composition according to claim 5, wherein step one comprises:
Adding deionized water into a stirring pot, heating to 80 ℃ under stirring, preserving heat for 30min, and cooling to 40 ℃.
7. A method of preparing an anti-aging repair composition according to claim 5, wherein step three comprises:
transferring the materials in the stirring pot into an ultrasonic equipment pot, performing ultrasonic treatment for 20min, and filtering and discharging at an ultrasonic frequency of 20kHz and 400 meshes to obtain the anti-aging repairing composition.
8. Use of an anti-aging treatment composition according to any one of claims 1 to 4 for the preparation of a skin care product comprising a cream, lotion, mask or essence.
9. Use of an anti-ageing repair composition according to claim 8 for the preparation of a skin care product, wherein the anti-ageing repair composition is present in the skin care product in an amount of 3% by mass.
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CN117122527A (en) * | 2023-07-04 | 2023-11-28 | 时垠(上海)生物科技有限公司 | Skin care composition, anti-aging cosmetic containing skin care composition and application of skin care composition |
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JP2010189363A (en) * | 2009-02-20 | 2010-09-02 | Seiwa Kasei Co Ltd | Cosmetic |
JP2011099020A (en) * | 2009-11-05 | 2011-05-19 | Seiwa Kasei Co Ltd | Antioxidant |
CN113876609A (en) * | 2021-10-29 | 2022-01-04 | 湖北省麦诗特生物科技有限公司 | Anti-aging composition containing hydroxypropyl tetrahydropyrane triol and polypeptide protein and application thereof |
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