Compositions and reagents and their applications, as well as promoters for the production of type I collagen and type III collagen and cosmetics

Abstract

The present invention relates to the technical field of cosmetics, and in particular to a composition, a reagent and their applications, as well as a promoter for the production of type I collagen and type III collagen and a cosmetic. The composition comprises 0.001 to 0.55 parts by weight of palmitoyl tripeptide trifluoroacetate, 0.001 to 1 part of methylsilanetriol, and 0.001 to 0.55 parts by weight of snake venom-like tripeptide. The technical solution provided by the present invention can not only regulate mitochondrial function, but also promote the expression of HSP47 protein, thereby promoting the production of collagen in the dermis, reducing the length and number of nasolabial folds and increasing skin elasticity, showing good anti-aging effects.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and particularly relates to a composition, a reagent and their applications, as well as a promoter for the production of type I collagen and type III collagen and a cosmetic product. Background Art

[0002] Collagen, also known as collagen, is a high-molecular-weight protein secreted by fibroblasts. It polymerizes into collagen fibrils outside cells and then forms collagen fibers through a small amount of cementing substance. The filamentous collagen fibers can keep the skin elastic. It exists in human skin, bones, teeth, tendons and other parts, and its main physiological function is to serve as an adhesive substance for connective tissues. In terms of the skin, it forms a reticular support structure with elastic fibers, providing a stable and strong support for the dermis. In human skin, the collagen content is as high as 71.9%, maintaining the elasticity and toughness of human skin. In terms of spatial structure, collagen is a transparent substance formed by a triple helix. Three independent collagen peptide chains rely on hydrogen bonds to maintain the structure of the triple helix wound around each other. So far, about 28 different types of collagen have been discovered, all with a triple helix wound structure.

[0003] If there is a lack of collagen in the human skin, it will lead to the cross-linking and solidification of collagen fibers and a reduction in intercellular mucopolysaccharides. As a result, the human skin will age and lose its elasticity, showing wrinkles, brown spots, sallow complexion, dryness, roughness, itching, aging, etc. Therefore, promoting the production of skin collagen is of great significance for maintaining healthy skin.

[0004] In view of this, the present invention is specifically proposed. Summary of the Invention

[0005] The purpose of the present invention is to solve the problem of how to promote the production of skin collagen, and provides a composition, a reagent and their applications, as well as a promoter for the production of type I collagen and type III collagen and a cosmetic product.

[0006] One object of the present invention is to provide a composition for promoting collagen production, which can promote the production of collagen by promoting the expression of the chaperone heat shock protein 47 (HSP47).

[0007] Another object of the present invention is to provide a reagent containing the aforementioned composition provided by the present invention.

[0008] Another object of the present invention is to provide an application of the aforementioned composition or reagent of the present invention in the preparation of a preparation for at least one of the following characteristics:

[0009] (a) Maintaining and / or increasing mitochondrial activity;

[0010] (b) Enhancing the expression of HSP47 protein;

[0011] (c) Enhance the expression of type I collagen and type III collagen.

[0012] Another object of the present invention is to provide a promoter for the production of type I collagen and type III collagen, which promoter comprises the foregoing composition or reagent provided by the present invention.

[0013] Another object of the present invention is to provide a cosmetic comprising the foregoing composition or reagent of the present invention.

[0014] In order to achieve the above objects of the present invention, on the one hand, the present invention provides a composition, wherein the composition comprises 0.001 to 0.55 parts by weight of palmitoyl tripeptide trifluoroacetate, 0.001 to 1 part of methylsilanetriol, and 0.001 to 0.55 parts by weight of snake venom-like tripeptide.

[0015] The composition provided by the present invention particularly has the function or effect of promoting collagen production.

[0016] In a specific embodiment of the present invention, in the composition, the mass ratio of palmitoyl tripeptide trifluoroacetate, methylsilanetriol, and snake venom-like tripeptide is (30 to 60):1:(10 to 40), and further is (40 to 50):1:(18 to 25).

[0017] On the other hand, the present invention provides a reagent which contains the composition of the present invention. Based on the total mass of the reagent, the mass content of the composition in the reagent is 0.003% to 2.1%.

[0018] In a specific embodiment of the present invention, the reagent further comprises at least one of glycerol, D-ribose, and amino acids.

[0019] In a specific embodiment of the present invention, the reagent comprises the following components by mass percentage: 0.001% to 0.55% of palmitoyl tripeptide trifluoroacetate, 0.001% to 1% of methylsilanetriol, 0.001% to 0.55% of snake venom-like tripeptide, 15% to 30% of glycerol, 5% to 20% of D-ribose, 0.5% to 3% of amino acids, and 44.9% to 79.497% of water.

[0020] On the other hand, the present invention provides the foregoing composition or reagent of the present invention for preparing a preparation, which preparation can increase the mitochondrial membrane potential of fibroblasts and can be applied to maintaining and / or increasing mitochondrial activity, enhancing HSP47 protein expression, or enhancing the expression of type I collagen and type III collagen.

[0021] In another aspect, the present invention provides a promoter for the production of type I collagen and type III collagen, which promoter comprises the composition or reagent described above in the present invention, and further the composition or reagent serves as an active ingredient in the promoter.

[0022] In yet another aspect, the present invention provides a cosmetic, which cosmetic comprises the composition or reagent described above in the present invention and cosmetically acceptable excipients.

[0023] In a specific embodiment of the present invention, based on the total mass of the cosmetic, the mass percentage of the composition or reagent is not more than 5%.

[0024] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0025] (1) The composition of the present invention combines palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom-like tripeptide in proportion. It can not only regulate mitochondrial function, but also promote the expression of HSP47 protein, thereby promoting the production of collagen in the dermis, reducing the length and number of nasolabial folds and increasing skin elasticity, showing good anti-aging effects;

[0026] (2) The composition of the present invention has good anti-aging effects and can be used as an active ingredient of cosmetics to be mixed with cosmetic excipients and the like to make various dosage forms of cosmetics, having practical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0028] Figure 1 Test results of the mitochondrial membrane potential of the compositions provided for different embodiments or comparative examples of the present invention;

[0029] Figure 2 Immunofluorescence images of HSP47 protein of the reagents provided for different embodiments or comparative examples of the present invention;

[0030] Figure 3 Comparative graph of the immunofluorescence intensity of HSP47 protein of the reagents provided for different embodiments or comparative examples of the present invention;

[0031] Figure 4 Comparative graph of the expression results of type I collagen of the reagents provided for different embodiments or comparative examples of the present invention;

[0032] Figure 5Comparison chart of type III collagen expression results of reagents provided in different embodiments or comparative examples of the present invention;

[0033] Figure 6 Comparison chart of the change in wrinkle depth before and after volunteers used the creams prepared from different embodiments or comparative examples of the present invention. Detailed implementation manners

[0034] The technical solutions of the present invention will be clearly and completely described below in conjunction with the accompanying drawings and specific implementation manners. However, those skilled in the art will understand that the following described embodiments are part of the embodiments of the present invention, rather than all embodiments, and are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the scope of protection of the present invention. For those conditions not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. For reagents or instruments not specified by the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0035] The biosynthesis of collagen fibers begins with the transcription of genes in the cell nucleus, is correctly folded and assembled into a triple helix structure in the endoplasmic reticulum, undergoes glycosylation and hydroxylation modifications, and then procollagen is secreted by the Golgi apparatus to the extracellular space for processing. Then, collagen aggregates into large fibers, becomes strengthened and assembled into bundles to form the final and insoluble collagen fibers. During this process, a collagen-specific molecular chaperone, heat shock protein 47 (HSP47), is involved. HSP47 is a protein located in the endoplasmic reticulum (protein synthesis chamber) of cells. It binds to procollagen in a neutral environment (endoplasmic reticulum) and dissociates in a slightly acidic environment (Golgi apparatus). HSP47 can modify procollagen, prevent the formation of aggregates of nascent procollagen chains, inhibit the degradation of procollagen in cells and the polymerization of procollagen triple helix structures, and can also promote the secretion of procollagen from the endoplasmic reticulum to the Golgi apparatus. In short, HSP47 is particularly important for the correct folding and assembly of the triple helix structure of procollagen. At the same time, there is a correlation between the overexpression of collagen and HSP47.

[0036] Based on this, the present invention uses palmitoyl tripeptide trifluoroacetate, methylsilanetriol, and snake venom-like tripeptide as the main active ingredients, and they are compounded in proportion to promote the expression of HSP47 protein, thereby achieving the promotion effect on collagen production.

[0037] On the one hand, the present invention provides a composition, which includes 0.001 - 0.55 parts by weight of palmitoyl tripeptide trifluoroacetate, 0.001 - 1 part by weight of methylsilanetriol, and 0.001 - 0.55 parts by weight of snake venom-like tripeptide. Preferably, it can promote collagen production.

[0038] The composition of the present invention is a compound of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom-like tripeptide in proportion. It can not only regulate mitochondrial function, but also promote the expression of HSP47 protein, thereby promoting the production of collagen in the dermis, reducing the length and number of nasolabial folds and increasing skin elasticity, showing good anti-aging effects. There is a mass ratio relationship among the above components of the composition provided by the present invention. The mass ratio of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom-like tripeptide is (0.1-55):(0.1-100):(0.1-55), so as to obtain the above technical effects.

[0039] Palmitoyl Tripeptide-5 (CAS: 623172-56-5) is a synthetic peptide composed of three amino acid chains and a fatty acid, belonging to the signal peptide. The structural formula is as follows:

[0040]

[0041] In palmitoyl tripeptide trifluoroacetate, the introduction of palmitoyl helps to better pass through the biological barrier and lipid membrane, enter the skin and exert its efficacy. In the composition of the present invention, by weight, the dosage of palmitoyl tripeptide trifluoroacetate can be 0.001 part, 0.005 part, 0.1 part, 0.2 part, 0.3 part, 0.4 part, 0.5 part, 0.55 part or the range composed of any two of them.

[0042] Methyl silanetriol (2445-53-6) is an organosilicon compound, a colorless liquid with a special odorless smell, and is usually used as a hair conditioner. The inventor of the present invention found in the research that the simultaneous introduction of methylsilanetriol when palmitoyl tripeptide trifluoroacetate and snake venom-like tripeptide are compounded provides a large number of binding sites for non-covalent bonds such as hydrogen bonds in the composition system. On the one hand, it improves the solubility of palmitoyl tripeptide trifluoroacetate and snake venom-like tripeptide, and on the other hand, it is more conducive to promoting transdermal absorption and then exerting efficacy. In the composition of the present invention, by weight, the dosage of methylsilanetriol can be 0.001 part, 0.005 part, 0.1 part, 0.2 part, 0.5 part, 0.6 part, 0.8 part, 1 part or the range composed of any two of them.

[0043] The tripeptide mimicking snake venom (CAS: 823202-99-9) is a small peptide that mimics the activity of snake venom toxins and can reduce wrinkles by inhibiting muscle contraction. However, the use of the tripeptide mimicking snake venom may cause symptoms such as skin redness and itching. In the present invention, through the compounding of three substances, by improving the transdermal absorption effect of the active ingredients, the efficacy can be exerted with a smaller addition amount, thus avoiding symptoms such as redness and itching. In the composition of the present invention, by weight, the dosage of the tripeptide mimicking snake venom can be 0.001 part, 0.005 part, 0.1 part, 0.2 part, 0.3 part, 0.4 part, 0.5 part, 0.55 part or the range composed of any two of them.

[0044] In a specific embodiment of the present invention, in the composition, the mass ratio of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and the tripeptide mimicking snake venom is (30-60):1:(10-40), and further is (40-50):1:(18-25).

[0045] The present invention further regulates the dosages of several substances in the composition, makes full use of the non-covalent interactions among palmitoyl tripeptide trifluoroacetate, methylsilanetriol and the tripeptide mimicking snake venom, improves the transdermal absorption effect of the active ingredients, and at the same time avoids the problems of product stability or storage stability caused by excessive non-covalent bonds, thereby affecting the use effect. For example, in different embodiments, in the composition, the mass ratio of palmitoyl tripeptide trifluoroacetate to methylsilanetriol can be 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1 or the range composed of any two of them; the mass ratio of methylsilanetriol to the tripeptide mimicking snake venom can be 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40 or the range composed of any two of them.

[0046] On the other hand, the present invention provides a reagent, which contains the composition of the present invention. Based on the total mass of the reagent, the mass content of the composition in the reagent is 0.003%-2.1%. That is, based on the total mass of the reagent, the sum of the mass percentages of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and the tripeptide mimicking snake venom is 0.003%-2.1%.

[0047] For example, in different embodiments, based on the total mass of the reagent, the sum of the mass percentages of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and the tripeptide mimicking snake venom can be 0.003%, 0.006%, 0.01%, 0.05%, 0.1%, 0.3%, 0.5%, 0.8%, 1%, 1.3%, 1.5%, 1.7%, 1.9%, 2.1% or the range composed of any two of them.

[0048] The reagent of the present invention can promote collagen production and may further include a cosmetically or pharmaceutically acceptable adjuvant, such as but not limited to an agent for improving cellular energy metabolism, an agent for improving or protecting mitochondrial function, an agent for promoting intercellular communication, a cell membrane protectant, an agent for increasing connexin in skin cells, an anti-aging agent, an anti-wrinkle agent, a skin tightening agent, a heat shock protein stimulant, an anti-chromatosis agent, a melanin synthesis stimulant or inhibitor, an antioxidant, an emulsifier, an emollient, a humectant, a moisture retention substance, a moisturizer, a vitamin, an amino acid, a protein, a thickening agent, a surfactant, a softening agent, an emulsifier, an adhesive, a preservative, a collagen synthesis stimulant, an elastin synthesis stimulant, a decorin synthesis stimulant, an agent for inhibiting collagen degradation, an agent for improving collagen fiber crosslinking, an agent for inhibiting matrix metalloproteinases, an agent for inhibiting elastin degradation, an agent for inhibiting serine proteases (such as kallikrein, elastase or cathepsin), an agent for stimulating fibroblast proliferation, an agent for stimulating keratinocyte proliferation, etc., provided that it is physically and chemically compatible with the remaining components of the reagent and specifically with the composition of the present invention. In addition, the nature of these additional components should not unacceptably alter the benefits of the composition of the present invention. The nature of these additional components can be synthetic or natural, or from a biotechnological process or from a combination of a synthetic process and a biotechnological process.

[0049] In some specific embodiments of the present invention, the present invention provides a reagent comprising palmitoyl tripeptide trifluoroacetate, methylsilanetriol and viper venom tripeptide as described above, and at least one of the following adjuvants in a pharmaceutically or cosmetically effective amount:

[0050] (1) A cosmetic or pharmaceutical adjuvant formed by a humectant, a moisture-retaining substance, a moisturizer and an emollient; (2) A compound that improves cellular energy metabolism, rejuvenates cells, enhances vitality, increases the energy supply level of cells or increases the energy level or adenosine triphosphate (ATP) production in cells; (3) A compound that can produce an anti-aging effect by regulating mitochondrial membrane potential, improves mitochondrial function with age, or prevents skin aging due to a decrease in mitochondrial membrane potential; (4) A compound having anti-wrinkle properties that improve skin conditions, or an agent that improves the appearance and vitality of the skin, thereby improving nasolabial folds, crow's feet, fatigue features, uneven skin tone and dryness; (5) A compound having the functions of promoting collagen synthesis, promoting elastin synthesis or inhibiting collagen degradation, inhibiting elastin degradation, inhibiting matrix metalloproteinase degradation; (6) Promote NAD + , an active substance that replenishes the consumption of NADase involved in physiological processes including DNA repair, metabolism and cell death.

[0051] In some specific embodiments of the present invention, the reagent further includes at least one of glycerol, D-ribose, and amino acids. Components such as glycerol, D-ribose, and amino acids can be added in conventional amounts, which will not be elaborated here.

[0052] Among them, glycerol serves as a skin moisturizer, and natural D-ribose and amino acids can further improve cellular energy metabolism and mitochondrial function. Amino acids include, but are not limited to, glutamic acid.

[0053] In addition, the reagent of the present invention may further include conventional preservatives such as phenoxyethanol, ethylhexylglycerol, etc.

[0054] In the specific embodiments of the present invention, the reagent includes the following components by mass percentage: palmitoyl tripeptide trifluoroacetate 0.001% - 0.55%, methylsilanetriol 0.001% - 1%, snake venom-like tripeptide 0.001% - 0.55%, glycerol 15% - 30%, ribose 5% - 20%, amino acids 0.5% - 3%, and water.

[0055] The preparation method of the reagent of the present invention is not limited. Just proportion the ingredients and mix them evenly.

[0056] The composition provided by the present invention can have a good effect of promoting collagen production. The corresponding mitochondrial membrane potential test, HSP47 protein expression test, type I and type III collagen expression test carried out by the further prepared above-mentioned reagent can prove the promoting effect of the composition of the present invention on collagen production.

[0057] On the other hand, the present invention provides an application of the aforementioned composition or reagent of the present invention in the preparation of a preparation for maintaining and / or increasing mitochondrial activity. Among them, the composition or reagent is used to increase the mitochondrial membrane potential of fibroblasts.

[0058] Mitochondria are the energy factories of cells, responsible for generating most of the energy required by cells. They synthesize ATP through the process of oxidative phosphorylation and participate in many other important functions of cells, including cell cycle regulation, apoptosis, calcium buffering, and antioxidant defense. During the aging process, mitochondria may be damaged, manifested as dysfunction. The decrease in mitochondrial membrane potential is usually closely related to mitochondrial dysfunction, and this state will further exacerbate the phenomenon of cell aging. The composition of the present invention can increase the mitochondrial membrane potential, promote autophagy of damaged mitochondria, thereby inhibiting the occurrence and development of cell aging.

[0059] On the other hand, the present invention provides an application of the aforementioned composition or reagent of the present invention in the preparation of a preparation for enhancing HSP47 protein expression.

[0060] On the other hand, the present invention provides an application of the aforementioned composition or reagent of the present invention in the preparation of a preparation for enhancing the expression of type I collagen and type III collagen.

[0061] Among the various collagens contained in vertebrates, type I collagen has the highest content, occupies most connective tissues, and is abundant in bones, corneas, tendons and dermis. The content of type I and type III fibrils determines the strength and elasticity of the skin. The composition obtained by the present invention from a certain ratio of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom-like tripeptide can promote the expression of HSP47 protein, thereby promoting the production of type I collagen and type III collagen, reducing the length and number of nasolabial folds and increasing skin elasticity, showing good anti-aging effects.

[0062] The application provided by the present invention can be used to prepare the aforementioned composition or reagent of the present invention into a preparation, which can be used for various potential application scenarios to promote the production of collagen in the dermis. As is well known in the art, the preparation generally also includes solvents, suspending agents, viscosity regulators, acid-base buffers, water-soluble cosolvents, plasticizers, moisturizers, disinfectants and antibacterial preservatives, etc., and the dosage is also the dosage well known in the art, which does not affect the functions that the composition or reagent provided by the present invention can achieve.

[0063] On another aspect, the present invention provides a promoter for the production of type I collagen and type III collagen, wherein the promoter contains the composition or reagent of the present invention as an active ingredient.

[0064] Among them, in the promoter for the production of type I collagen and type III collagen, the above composition or reagent can be used as the only active ingredient for promoting the production of type I collagen and type III collagen.

[0065] On another aspect, the present invention provides an application of any one of the aforementioned compositions or reagents of the present invention in the preparation of cosmetics.

[0066] In the specific embodiments of the present invention, the cosmetics include at least one of essence, lotion, cream and mask. The state of the cosmetics of the present invention is not limited thereto and can be adjusted conventionally according to actual needs.

[0067] On another aspect, the present invention provides a cosmetic, which includes any one of the above compositions or reagents, and cosmetically acceptable excipients.

[0068] In the specific embodiments of the present invention, based on the total mass of the cosmetic, the mass percentage of the composition or reagent is not more than 5%. Preferably, it is 0.1% - 5%.

[0069] In different embodiments, based on the total mass of the cosmetic, the mass percentage of the composition can be 0.1%, 0.3%, 0.5%, 0.8%, 1%, 1.5%, 2%, 3%, 4%, 5%, or a range composed of any two of them.

[0070] In a specific embodiment of the present invention, the excipients include at least one of emollients, emulsifiers, thickeners, humectants, skin protectants, and preservatives. The usage amount can be well-known in the art and is an amount that does not affect the efficacy of the composition or reagent.

[0071] The materials used in the following examples, comparative examples, and experimental examples are all commercially available as usual.

[0072] Example 1

[0073] This example provides a reagent containing a composition for promoting collagen production, including the following components by mass percentage: palmitoyl tripeptide trifluoroacetate 0.55%, methylsilanetriol 1%, snake venom-like tripeptide 0.55%, glycerol 23.4%, D-ribose 10.42%, glutamic acid 0.65%, and the balance water.

[0074] The preparation method of the reagent in this example includes: weighing each component according to the ratio and stirring and mixing evenly at room temperature.

[0075] Example 2

[0076] This example provides a reagent containing a composition for promoting collagen production, including the following components by mass percentage: palmitoyl tripeptide trifluoroacetate 0.001%, methylsilanetriol 0.001%, snake venom-like tripeptide 0.001%, glycerol 23.4%, D-ribose 10.42%, glutamic acid 0.65%, and the balance water.

[0077] The preparation method of the reagent in this example includes: weighing each component according to the ratio and stirring and mixing evenly at room temperature.

[0078] Example 3

[0079] This example provides a reagent containing a composition for promoting collagen production, including the following components by mass percentage: palmitoyl tripeptide trifluoroacetate 0.23%, methylsilanetriol 0.005%, snake venom-like tripeptide 0.11%, glycerol 23.4%, D-ribose 10.42%, glutamic acid 0.65%, and adding the balance water until the sum of the contents of each component is 100%.

[0080] The preparation method of the reagent in this example includes: weighing each component according to the ratio and stirring and mixing evenly at room temperature.

[0081] Example 4

[0082] This embodiment provides a reagent containing a composition that promotes collagen production, including the following components by mass percentage: palmitoyl tripeptide trifluoroacetate 0.091%, methylsilanetriol 0.165%, snake venom-like tripeptide 0.091%, glycerol 23.4%, D-ribose 10.42%, glutamic acid 0.65%, and the balance being water.

[0083] The preparation method of the reagent in this embodiment includes: weighing each component in proportion and stirring and mixing evenly at room temperature.

[0084] Examples 5 to 8

[0085] This embodiment provides a facial cream, and the formula is shown in Table 1.

[0086] Table 1 Different facial cream formulas (wt%)

[0087]

[0088] The preparation method of the facial cream in this embodiment includes the following steps:

[0089] (1) Heat the deionized water in phase A to 80 - 85°C;

[0090] (2) Add the components in phase B in the order of glyceryl stearate, glyceryl stearate / PEG - 100 stearate complex, isocetane, isooctyl palmitate, cetearyl alcohol to the oil pan, pre - mix and heat to 80 - 85°C, and stir evenly;

[0091] (3) Add the material obtained in step (2) to the material obtained in step (1) with homogenization turned on, stir and mix evenly and homogenize and emulsify for 10 minutes;

[0092] (4) Cool the material obtained in step (3) to below 40°C, and add the components of phase C in the order of phenoxyethanol, SC50 ethylhexylglycerin, and the reagent of this embodiment respectively, stir evenly, and cool to room temperature.

[0093] Comparative Examples 1 to 6

[0094] Comparative Examples 1 to 6 provide different reagents, and the specific compositions are shown in Table 2. The preparation method of the reagent includes: weighing each component in proportion and stirring and mixing evenly at room temperature.

[0095] Table 2 Different composition compositions (wt%)

[0096]

[0097] Comparative Example 7

[0098] Comparative Example 7 provides a cream, referring to the cream of Example 5 and its preparation method, the difference being that: the reagent of Comparative Example 4 with equal weight is used to replace the reagent of Example 1.

[0099] Comparative Example 8

[0100] Comparative Example 8 provides a cream, referring to the cream of Example 5 and its preparation method, the difference being that: the reagent of Example 1 is not added, and the amount of the missing reagent of Example 1 is supplemented with deionized water.

[0101] Experimental Example 1

[0102] The mitochondrial membrane potential of the reagents of Examples 1-4 and Comparative Examples 1-6 was tested. In this experiment, fibroblasts were used as the object to evaluate the effect of each reagent on the mitochondrial membrane potential. The specific experimental steps are as follows.

[0103] Cell seeding: Human fibroblasts with a confluence of 80% were plated, with a volume of 1 mL per well, and plated in a 12-well plate to make the cell density reach 7×10 4 ~9×10 4 / well.

[0104] Observation of cells: The cell state is good, and the number is appropriate, and the next experiment can be carried out.

[0105] Setting groups: Blank group, model group, Example 1, Example 2, Example 3, Example 4, Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Comparative Example 5, Comparative Example 6.

[0106] Model culture: Discard the culture medium, wash with PBS, and then add CCCP (model group) for incubation.

[0107] Sample addition and culture: On the basis of the model group, discard the culture medium, wash with PBS, and then add 1 mL / well of high-glucose DMEM complete medium (blank group and model group), 1 mL / well of high-glucose DMEM complete medium containing the composition of the example / comparative example (composition group, the content of the composition in the high-glucose DMEM complete medium is 1 wt%), and then place the well plate in an incubator at 37°C and 5% CO2 for 24 h.

[0108] Cell collection: Use trypsin to digest the cells in the well plate, centrifuge and discard the supernatant, add mitochondrial membrane potential detection solution (JC-1), and incubate. Then wash with pre-cooled buffer, centrifuge 3 times repeatedly, and discard the supernatant. Collect the cells into a new EP tube and add fresh pre-cooled buffer.

[0109] Mitochondrial fluorescence measurement: Take 100 uL of cell solution into a 96-well plate, and detect the fluorescence intensities of JC-1 monomer and polymer respectively, and perform ratio analysis.

[0110] Figure 1 Test results of the mitochondrial membrane potential of the reagents provided for different embodiments or comparative examples of the present invention. Mitochondria in cells, as the main site for aerobic respiration to produce ATP, are one of the important organelles in the body. Mitochondria use oxidizable substrates to generate an electrochemical proton gradient on the mitochondrial membrane, and this gradient is used to produce ATP. Under normal circumstances, the membrane potential of mitochondria shows positive outside and negative inside. During the process of mitochondrial dysfunction, the mitochondrial membrane potential will decrease, which may then cause the opening of mitochondrial permeability pores and the loss of the electrochemical gradient. Therefore, the mitochondrial membrane potential is an essential parameter of mitochondrial function and can be used as an indicator of cell health. JC-1 is an ideal fluorescent probe widely used to detect mitochondrial membrane potential. When the mitochondrial membrane potential is relatively high, JC-1 aggregates in the mitochondrial matrix to form polymers, which can produce red fluorescence; when the mitochondrial membrane potential is relatively low, JC-1 cannot aggregate in the mitochondrial matrix, and at this time JC-1 is a monomer, which can produce green fluorescence. Therefore, the ratio of red to green fluorescence (Red / Green) is used to measure the proportion of mitochondrial depolarization. The lower the Red / Green ratio, the lower the mitochondrial membrane potential; the higher the Red / Green ratio, the higher the mitochondrial membrane potential. Therefore, the active substance has the effect of increasing the mitochondrial membrane potential, improving mitochondrial function, and restoring cell health. As can be seen from Figure 1 it, the mitochondrial membrane potential of fibroblasts in the model group was significantly decreased, and the mitochondrial membrane potential measured by the reagents of Examples 1 to 4 was significantly increased, with a statistical difference of p < 0.01. Among them, the mitochondrial membrane potential measured in Example 3 was higher and the effect was better; the mitochondrial membrane potential measured by the reagents of Comparative Examples 1 to 6 did not change significantly.

[0111] Experimental Example 2

[0112] The reagents of Examples 1 to 4 and Comparative Examples 1 to 6 were tested for HSP47 protein expression. In this experiment, fibroblasts were used as the object to evaluate the effect of each reagent on HSP47 protein expression. The specific experimental steps are as follows.

[0113] Cell seeding: Human fibroblasts with a confluence of 80% were plated, with a volume of 2 mL per well, and plated in a 6-well plate to make the cell density reach 20×10 4 / well.

[0114] Group setting: blank group, model group, Example 1, Example 2, Example 3, Example 4, Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Comparative Example 5, Comparative Example 6.

[0115] Model culture: Discard the culture medium, wash with PBS, and then carry out culture under heat stimulation conditions (model group).

[0116] Sample addition and culture: Discard the culture medium, wash with PBS, and add 2 mL / well of high-glucose DMEM complete medium (blank group and model group), 2 mL / well of high-glucose DMEM complete medium containing the reagent of the example / comparative example (reagent group, the content of the reagent in the high-glucose DMEM complete medium is 1 wt%). Subsequently, establish the model, and then place the well plate in an incubator at 37 °C and 5% CO2 for 24 h.

[0117] HSP47 immunofluorescence detection: Discard the culture medium, wash once or twice with PBS, perform fixation, permeabilization, and blocking processes, then incubate with the primary antibody and the secondary antibody, and perform microscopic observation and fluorescence intensity data analysis (total fluorescence intensity / cell number).

[0118] The immunofluorescence images and fluorescence intensity comparison charts of the HSP47 protein of the reagents provided by different examples or comparative examples are respectively as Figure 2 and Figure 3 shown. From Figure 2 , 3 it can be seen that there is no statistical difference in the expression of HSP47 protein in fibroblasts between the model group and the blank group. The expression of HSP47 measured by the reagents of Examples 1 to 4 increased, with a statistical difference of p < 0.05. Among them, the expression effect measured by Example 3 was better, and the expression of HSP47 measured by the reagents of Comparative Examples 1 to 6 did not increase significantly.

[0119] Experimental Example 3

[0120] Test the expression of type I and type III collagen of the reagents of Examples 1 to 4 and Comparative Examples 1 to 6. In this experiment, fibroblasts were used as the object to evaluate the effects of each reagent on the expression of type I and type III collagen. The specific experimental steps are as follows.

[0121] Cell seeding: Plate the human fibroblasts with a confluence of 80%, with a volume of 2 mL per well, and plate them in a 24-well plate to make the cell density reach 4×10 4 / well.

[0122] Set the groups: blank group, model group, Example 1, Example 2, Example 3, Example 4, Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Comparative Example 5, Comparative Example 6.

[0123] Sample addition and culture: Discard the culture medium, wash with PBS, and add high-glucose DMEM complete medium (blank group), high-glucose DMEM complete medium containing the reagent of the example / comparative example (reagent group, the content of the reagent in the high-glucose DMEM complete medium is 0.1 wt%), high-glucose DMEM complete medium containing the positive control (0.01 wt% ascorbic acid AA) (positive control group). Then place the well plate in an incubator at 37 °C and 5% CO2 for 72 h.

[0124] Immunofluorescence detection of type I and type III collagen: Discard the culture medium, wash once or twice with PBS, and after the fixation, permeabilization, and blocking processes, incubate with type I or type III primary antibody and secondary antibody respectively, then observe under a microscope and analyze the fluorescence intensity data (total fluorescence intensity / cell number). Use Image J software to measure the total fluorescence intensity in the same area, keep the threshold consistent, calculate the number of cells in the same area, and the average fluorescence intensity = total fluorescence intensity / cell number. The final result takes the blank group as 100%, and calculates the relative content of each group.

[0125] The comparison charts of type I collagen expression results and type III collagen expression results of the reagents provided by different embodiments or comparative examples of the present invention are respectively as Figure 4 and Figure 5 shown. It can be seen from Figure 4 , 5 that the reagents in the positive control group and Examples 1 to 4 all increased the contents of type I and type III collagen, and the statistical difference p < 0.05. Among them, the implementation effect of Example 3 is better; the reagents in Comparative Examples 1 to 6 have no significant improvement on collagen.

[0126] Experimental Example 4

[0127] Conduct a volunteer use experiment on the creams of Examples 5 to 8 and Comparative Examples 7 to 8.

[0128] Experimental steps: Recruit a number of volunteers aged between 30 and 60 years old with mild, severe wrinkles and skin laxity on the face. After collecting basic data (the depth of nasolabial folds and skin elasticity), perform radiofrequency treatment. Randomly, 3 people use the cream of Comparative Example 8 (placebo), 3 people use the cream of Comparative Example 7, 3 people use the cream of Example 5, 3 people use the cream of Example 6, 3 people use the cream of Example 7, and 3 people use the cream of Example 8, once in the morning and once in the evening every day. Make a follow-up visit at the 8th week and collect the above skin data. The test results are shown in Table 3, where:

[0129] The difference in wrinkle depth relative to the baseline value = wrinkle depth (baseline value) - wrinkle depth (after use);

[0130] The change rate of skin elasticity relative to the baseline value % = [skin elasticity (after use) - skin elasticity (baseline value)] / skin elasticity (baseline value) × 100%.

[0131] Table 3 Changes in wrinkle depth and skin elasticity of users of different creams

[0132]

[0133] Table 3 shows that for Example 7, the numerical value of the difference in wrinkle depth relative to the baseline value and the rate of change in skin elasticity relative to the baseline value are the largest, indicating that the anti-wrinkle effect implemented on the skin is the best.

[0134] Figure 6 This is a comparison chart of the change in wrinkle depth before and after volunteers used the creams prepared from different examples or comparative examples of the present invention. From the above test results, it can be seen that compared with the placebo cream of Comparative Example 8 and the baseline value, after using the cream of the example for 8 weeks, the average depth of skin texture has significantly decreased, and the skin elasticity has significantly increased. Moreover, Example 7 has the best improvement effect on wrinkle depth and elasticity (the green shaded part shown in the image is the smallest. Image after 8 weeks). There is no significant difference before and after using Comparative Example 7.

[0135] In summary, the composition or reagent of the present invention can promote the expression of HSP47 protein, promote the production of type I and type III collagen, and it is found in human tests that after using the cream containing the corresponding reagent for 8 weeks, the depth of nasolabial folds has significantly decreased and the skin elasticity has significantly increased, which further demonstrates that the composition or reagent of the present invention can promote collagen production.

[0136] 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 it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A promoter composition for the production of type I collagen and type III collagen, characterized in that, The composition consists of 0.001 to 0.55 parts by weight of palmitoyl tripeptide trifluoroacetate, 0.001 to 1 part of methylsilanetriol, and 0.001 to 0.55 parts of snake venom-like tripeptide; In the composition, the mass ratio of palmitoyl tripeptide trifluoroacetate, methylsilanetriol, and snake venom-like tripeptide is (30 - 60) : 1 : (10 - 40).

2. The composition according to claim 1, wherein In the composition, the mass ratio of palmitoyl tripeptide trifluoroacetate, methylsilanetriol, and snake venom-like tripeptide is (40 - 50) : 1 : (18 - 25).

3. A reagent, characterized in that, The reagent contains the composition described in any one of claims 1 - 2. Based on the total mass of the reagent, the mass content of the composition in the reagent is 0.003% - 2.1%.

4. The reagent according to claim 3, characterized in that, The reagent further includes at least one of glycerol, D-ribose, and amino acids.

5. The reagent according to claim 4, wherein The reagent includes the following components by mass percentage: 0.001% - 0.55% of palmitoyl tripeptide trifluoroacetate, 0.001% - 1% of methylsilanetriol, 0.001% - 0.55% of snake venom-like tripeptide, 15% - 30% of glycerol, 5% - 20% of D-ribose, 0.5% - 3% of amino acids, and 44.9% - 79.497% of water.

6. Use of the composition described in any one of claims 1 - 2 or the reagent described in any one of claims 3 - 5 in the preparation of a preparation for anti-aging.

7. A cosmetic, characterized in that, The cosmetic includes the composition described in any one of claims 1 - 2 or the reagent described in any one of claims 3 - 5 and cosmetically acceptable excipients.

8. The cosmetic according to claim 7, characterized in that, Based on the total mass of the cosmetic, the mass percentage of the composition or the reagent is not more than 5%.

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