Composition, reagent, application of composition and reagent, generation accelerant of type I collagen and type III collagen and cosmetic

Abstract

The invention relates to the technical field of cosmetics, in particular to a composition, a reagent, application of the composition and the reagent, a generation accelerant of type I collagen and type III collagen and cosmetics. The composition is prepared from the following components in parts by weight: 0.001 to 0.55 part of palmitoyl tripeptide trifluoroacetate, 0.001 to 1 part of methylsilanetriol and 0.001 to 0.55 part of snake venom-like tripeptide. According to the technical scheme provided by the invention, not only can the mitochondrial function be adjusted, but also the expression of HSP47 protein can be promoted, so that the collagen generation of dermis is promoted, the length and the quantity of nasolabial folds are reduced, the skin elasticity is improved, and a good anti-aging effect is shown.

Description

Technical Field

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

[0002] Collagen, also known as collagen, is a high molecular protein secreted by fibroblasts, polymerized into collagen fibrils outside the cells, and then bonded into collagen fibers by a small amount of adhesive. Filamentous collagen fibers can keep the skin elastic. It exists in human skin, bones, teeth, tendons and other parts. Its main physiological function is to serve as an adhesive substance for connective tissue. In the skin, it forms a mesh support structure with elastic fibers, providing 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 triple helices. Three independent collagen peptide chains rely on hydrogen bonds to maintain the structure of the triple helices intertwined with each other. So far, about 28 different types of collagen have been discovered, all of which have a triple helical winding structure.

[0003] Lack of collagen in human skin will lead to cross-linking and solidification of collagen fibers, and reduction of intercellular mucopolysaccharides. Human skin will age and lose elasticity, and wrinkles, brown spots, chlorosis, 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 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 to provide a composition and reagent and their application, as well as a production promoter and cosmetics for type I collagen and type III collagen.

[0006] One object of the present invention is to provide a composition for promoting collagen production, which can promote collagen production by promoting the expression of 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 a use of the aforementioned composition or agent of the present invention in preparing a preparation having at least one of the following characteristics: (a) maintain and / or increase mitochondrial activity; (b) Enhanced HSP47 protein expression; (c) Enhanced the expression of type I and type III collagen.

[0009] Another object of the present invention is to provide a promoter for the production of type I collagen and type III collagen, wherein the promoter comprises the aforementioned composition or the aforementioned reagent provided by the present invention.

[0010] Another object of the present invention is to provide a cosmetic comprising the aforementioned composition or the aforementioned agent of the present invention.

[0011] In order to achieve the above-mentioned purpose of the present invention, the present invention provides a composition on the one hand, wherein the composition comprises 0.001 to 0.55 parts of palmitoyl tripeptide trifluoroacetate, 0.001 to 1 parts of methylsilanetriol and 0.001 to 0.55 parts of snake venom tripeptide in parts by weight.

[0012] The composition provided by the present invention has a function or effect of promoting collagen production.

[0013] 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 (30-60) : 1 : (10-40), further (40-50) : 1 : (18-25).

[0014] Another aspect of the present invention provides a reagent, wherein the reagent contains the composition of the present invention, and the mass content of the composition in the reagent is 0.003% to 2.1% based on the total mass of the reagent.

[0015] In a specific embodiment of the present invention, the reagent further comprises at least one of glycerol, D-ribose and amino acid.

[0016] In a specific embodiment of the present invention, the reagent includes the following components by mass percentage: 0.001% to 0.55% palmitoyl tripeptide trifluoroacetate, 0.001% to 1% methylsilanetriol, 0.001% to 0.55% snake venom tripeptide, 15% to 30% glycerol, 5% to 20% D-ribose, 0.5% to 3% amino acids and 44.9% to 79.497% water.

[0017] On the other hand, the present invention provides the aforementioned composition or reagent of the present invention for preparing a preparation, wherein the preparation can increase the mitochondrial membrane potential of fibroblasts, and can be used to maintain and / or increase mitochondrial activity, enhance HSP47 protein expression, or enhance the expression of type I collagen and type III collagen.

[0018] In another aspect, the present invention provides an agent for promoting the production of type I collagen and type III collagen, wherein the agent comprises the aforementioned composition or reagent of the present invention, and the composition or reagent serves as an active ingredient in the agent.

[0019] In another aspect, the present invention provides a cosmetic comprising the aforementioned composition or reagent of the present invention and cosmetically acceptable auxiliary materials.

[0020] In a specific embodiment of the present invention, the mass percentage of the composition or agent is no more than 5% based on the total mass of the cosmetic.

[0021] Compared with the prior art, the present invention has the following beneficial effects: (1) The composition of the present invention is a mixture of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom tripeptide in a certain proportion, which 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; (2) The composition of the present invention has good anti-aging effects and can be used as an effective ingredient of cosmetics and mixed with cosmetic excipients to prepare cosmetics of various dosage forms, thus having practical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0023] Figure 1 The test results of mitochondrial membrane potential of the compositions provided in different embodiments or comparative examples of the present invention; Figure 2 Immunofluorescence images of HSP47 proteins of reagents provided in different embodiments or comparative examples of the present invention; Figure 3 A comparison chart of the immunofluorescence intensity of HSP47 protein of reagents provided in different embodiments or comparative examples of the present invention; Figure 4 A comparison chart of the expression results of type I collagen of reagents provided in different embodiments or comparative examples of the present invention; Figure 5 A comparison chart of the expression results of type III collagen of reagents provided in different embodiments or comparative examples of the present invention; Figure 6 The following is a comparison chart of the changes in wrinkle depth of volunteers before and after using the facial creams prepared from different embodiments or comparative examples of the present invention. DETAILED DESCRIPTION

[0024] The technical scheme of the present invention will be clearly and completely described below in conjunction with the accompanying drawings and specific embodiments, but it will be understood by those skilled in the art that the following described embodiments are part of embodiments of the present invention, rather than all embodiments, and are only used to illustrate the present invention, and should not be considered 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 making creative work are within the scope of protection of the present invention. If specific conditions are not specified in the embodiments, they are carried out according to normal conditions or conditions recommended by the manufacturer. If the manufacturer is not specified in the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0025] The biosynthesis of collagen fibers begins with the transcription of genes in the cell nucleus, and is correctly folded and assembled into a triple helical structure in the endoplasmic reticulum. After glycosylation and hydroxylation, procollagen is secreted from the Golgi apparatus to the extracellular space for processing, and then the collagen aggregates into large fibers, becomes strengthened and assembled into bundles, and forms the final and insoluble collagen fibers. In 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 the cell. It binds to procollagen in a neutral environment (endoplasmic reticulum) and dissociates in an acidic environment (Golgi apparatus). HSP47 can modify procollagen and prevent the formation of aggregates of newly formed procollagen chains, inhibit the degradation of procollagen in cells and the polymerization of the procollagen triple helical structure, 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 helical structure of procollagen. Meanwhile, there was a correlation between the overexpression of collagen and HSP47.

[0026] Based on this, the present invention uses palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom-like tripeptide as main active ingredients, which are compounded in proportion to promote the expression of HSP47 protein, thereby achieving the effect of promoting collagen production.

[0027] In one aspect, the present invention provides a composition, which comprises, by weight, 0.001 to 0.55 parts of palmitoyl tripeptide trifluoroacetate, 0.001 to 1 parts of methylsilanetriol, and 0.001 to 0.55 parts of snake venom-like tripeptide, which can preferably promote collagen production.

[0028] The composition of the present invention mixes palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom-like tripeptide in proportion, which can not only regulate mitochondrial function, but also promote the expression of HSP47 protein, thereby promoting collagen production 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 between the above components of the composition provided by the present invention, and the mass ratio of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom-like tripeptide is (0.1-55): (0.1-100): (0.1-55), thereby obtaining the above technical effects.

[0029] Palmitoyl Tripeptide-5 (CAS: 623172-56-5) is a synthetic peptide composed of three amino acid chains and fatty acids. It belongs to the signal peptide class and has the following structural formula:

[0030] In palmitoyl tripeptide trifluoroacetate, the access of palmitoyl helps to better pass through biological barriers and lipid membranes, enter the skin, and exert its efficacy. In the composition of the present invention, the amount of palmitoyl tripeptide trifluoroacetate can be 0.001 parts, 0.005 parts, 0.1 parts, 0.2 parts, 0.3 parts, 0.4 parts, 0.5 parts, 0.55 parts, or a range consisting of any two of them, by weight.

[0031] Methyl silanetriol (2445-53-6) is an organic silicon compound, a colorless liquid, with a special smell without peculiar odor, and is usually used as a hair conditioner. The inventors of the present invention found in their research that the introduction of methyl silanetriol while compounding palmitoyl tripeptide trifluoroacetate and snake venom-like tripeptide provides a large number of binding sites for non-covalent bonds such as hydrogen bonds for the composite system, which improves the solubility of palmitoyl tripeptide trifluoroacetate and snake venom-like tripeptide on the one hand, and helps promote transdermal absorption on the other hand, thereby exerting the efficacy. In the composition of the present invention, the amount of methyl silanetriol can be 0.001 parts, 0.005 parts, 0.1 parts, 0.2 parts, 0.5 parts, 0.6 parts, 0.8 parts, 1 part, or a range consisting of any two of them, by weight.

[0032] Snake venom tripeptide (CAS: 823202-99-9) is a small peptide that simulates the activity of snake venom toxins. It can reduce wrinkles by inhibiting muscle contraction, but the use of snake venom tripeptide may cause skin redness, swelling, itching and other symptoms. The present invention improves the transdermal absorption effect of the active ingredients by compounding the three substances, so that the efficacy can be exerted at a smaller addition amount, avoiding symptoms such as redness, swelling, itching and the like. In the composition of the present invention, the amount of snake venom tripeptide can be 0.001 parts, 0.005 parts, 0.1 parts, 0.2 parts, 0.3 parts, 0.4 parts, 0.5 parts, 0.55 parts or the range of any two of them by weight.

[0033] 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 (30-60) : 1 : (10-40), further (40-50) : 1 : (18-25).

[0034] The present invention further regulates the dosage of several substances in the composition, makes full use of the non-covalent interaction between palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom-like tripeptide, improves the transdermal absorption effect of the active ingredient, and avoids the product stability or storage stability problems caused by too many 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 of any two thereof; the mass ratio of methylsilanetriol to snake venom-like tripeptide can be 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40 or the range of any two thereof.

[0035] Another aspect of the present invention provides a reagent, the reagent contains the composition of the present invention, and the mass content of the composition in the reagent is 0.003% to 2.1% based on the total mass of the reagent. That is, based on the total mass of the reagent, the sum of the mass percentages of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom-like tripeptide is 0.003% to 2.1%.

[0036] 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 snake venom-like tripeptide 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 a range consisting of any two of them.

[0037] The agent of the present invention can promote collagen production, and may further include cosmetically or pharmaceutically acceptable adjuvants, such as but not limited to agents for improving cell energy metabolism, agents for improving or protecting mitochondrial function, agents for promoting intercellular communication, cell membrane protectants, agents for increasing connexins in skin cells, anti-aging agents, anti-wrinkle agents, skin tightening agents, heat shock protein stimulators, anti-pigmentation agents, melanin synthesis stimulators or inhibitors, antioxidants, emulsifiers, emollients, humectants, moisture retaining substances, moisturizers, vitamins, amino acids, proteins, thickeners, surfactants The invention also provides the invention claims a kind of preparation method of the present invention.The preparation method of the present invention can be used for the preparation of the present invention.For example, the preparation of the present invention comprises the following: the preparation of the present invention comprises the following: , softening agent, emulsifying agent, adhesive, preservative, collagen synthesis stimulator, elastin synthesis stimulator, decorin synthesis stimulator, the medicament that inhibits collagen degradation, the medicament that improves collagen fiber cross-linking, the medicament that inhibits matrix metalloproteinase, the medicament that inhibits elastin degradation, the medicament that inhibits serine protease (such as kallikrein, elastase or cathepsin), the medicament that stimulates fibroblast proliferation, the medicament that stimulates keratinocyte proliferation etc., condition is that it is physically and chemically compatible with the remaining components of the reagent and particularly with the composition of the present invention.In addition, the property of these other ingredients should not unacceptably change the benefit of the present composition.The property of these other ingredients can be synthetic or natural, or from biotechnological process or from the combination of synthesis procedure and biotechnological process.

[0038] In some specific embodiments of the present invention, the present invention provides a reagent comprising the above palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom tripeptide, and at least one of the following adjuvants in a pharmaceutically or cosmetically effective amount: (1) Cosmetic or pharmaceutical adjuvants formed from moisturizers, moisture-retaining substances, humectants and emollients; (2) Compounds that improve cellular energy metabolism, rejuvenate cells, enhance vitality, increase cellular energy levels or increase energy levels or adenosine triphosphate (ATP) production in cells; (3) Compounds that can produce anti-aging effects by regulating mitochondrial membrane potential, improve mitochondrial function with age, or prevent skin aging due to decreased mitochondrial membrane potential; (4) Compounds with anti-wrinkle properties that improve skin condition, or agents that improve the appearance and vitality of the skin and thus improve nasolabial folds, crow's feet, fatigue characteristics, uneven skin tone and dryness; (5) Compounds that promote collagen synthesis, promote elastin synthesis or inhibit collagen degradation, inhibit elastin degradation, and inhibit matrix metalloproteinase degradation; (6) Compounds that promote NAD + , to replenish the depleted active substances of NADase involved in physiological processes including DNA repair, metabolism and cell death.

[0039] In some specific embodiments of the present invention, the reagent further comprises at least one of glycerol, D-ribose and amino acid. Glycerol, D-ribose, amino acid and other components can be added in conventional amounts, which will not be described in detail here.

[0040] Among them, glycerol acts as a skin moisturizer, and natural D-ribose and amino acids can further improve cell energy metabolism and mitochondrial function. Amino acids include but are not limited to glutamic acid.

[0041] In addition, the agent of the present invention may also include conventional preservatives such as phenoxyethanol, ethylhexylglycerin, and the like.

[0042] In a specific embodiment of the present invention, the reagent includes the following components by mass percentage: 0.001% to 0.55% palmitoyl tripeptide trifluoroacetate, 0.001% to 1% methylsilanetriol, 0.001% to 0.55% snake venom tripeptide, 15% to 30% glycerol, 5% to 20% ribose, 0.5% to 3% amino acids and water.

[0043] The preparation method of the reagent of the present invention is not limited, and the ingredients are mixed in proportion and evenly mixed.

[0044] The composition provided by the present invention can effectively promote collagen production, and the promoting effect of the composition of the present invention on collagen production can be demonstrated by the corresponding mitochondrial membrane potential test, HSP47 protein expression test, type I and type III collagen expression test performed by the above-mentioned reagents further prepared.

[0045] Another aspect of the present invention provides a use of the aforementioned composition or reagent of the present invention in the preparation of a preparation for maintaining and / or increasing mitochondrial activity, wherein the composition or reagent is used to increase the mitochondrial membrane potential of fibroblasts.

[0046] Mitochondria are the energy factories of cells, responsible for producing 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 and manifest as dysfunction. The reduction of mitochondrial membrane potential is usually closely related to mitochondrial dysfunction, which further aggravates the phenomenon of cell aging. The composition of the present invention can enhance the mitochondrial membrane potential and promote autophagy of damaged mitochondria, thereby inhibiting the occurrence and development of cell aging.

[0047] Another aspect of the present invention provides a use of the aforementioned composition or reagent of the present invention in the preparation of a preparation for enhancing the expression of HSP47 protein.

[0048] Another aspect of the present invention provides a use 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.

[0049] Among the various collagens contained in vertebrates, type I collagen has the highest content, occupies most of the connective tissues, and is more 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 through a certain ratio of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom tripeptide can achieve the effect of promoting the production of type I collagen and type III collagen by promoting the expression of HSP47 protein, reducing the length and number of nasolabial folds and increasing skin elasticity, showing good anti-aging effect.

[0050] The application provided by the present invention can realize the preparation of the aforementioned composition or reagent of the present invention into a preparation, which can be used to realize various potential application scenarios for promoting collagen production in the dermis. The preparation can also be prepared as known in the art, generally with solvents, suspending agents, viscosity modifiers, acid-base buffers, water-soluble cosolvents, plasticizers, moisturizers, disinfectants and antibacterial preservatives, etc., and the dosage is also a dosage known in the art, which does not affect the functions that can be achieved by the composition or reagent provided by the present invention.

[0051] In another aspect, the present invention provides an agent for promoting the production of type I collagen and type III collagen, wherein the agent contains the composition or reagent of the present invention as an effective ingredient.

[0052] Among them, in the agent for promoting the production of type I collagen and type III collagen, the above composition or reagent can be used as the only effective ingredient for promoting the production of type I collagen and type III collagen.

[0053] Another aspect of the present invention provides use of any one of the above-mentioned compositions or reagents of the present invention in the preparation of cosmetics.

[0054] In a specific embodiment 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 conventionally adjusted according to actual needs.

[0055] In another aspect, the present invention provides a cosmetic comprising any one of the above compositions or agents, and cosmetically acceptable auxiliary materials.

[0056] In a specific embodiment of the present invention, the mass percentage of the composition or agent is no more than 5% based on the total mass of the cosmetic, preferably 0.1% to 5%.

[0057] For example, 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 consisting of any two thereof.

[0058] In a specific embodiment of the present invention, the auxiliary material includes at least one of an emollient, an emulsifier, a thickener, a moisturizer, a skin protectant and a preservative. The amount used can be an amount known in the art and does not affect the efficacy of the composition or agent.

[0059] The materials used in the following examples, comparative examples and experimental examples are all commercially available.

[0060] Example 1 This embodiment provides a reagent containing a composition for promoting collagen production, including the following components in mass percentage: 0.55% palmitoyl tripeptide trifluoroacetate, 1% methylsilanetriol, 0.55% snake venom tripeptide, 23.4% glycerol, 10.42% D-ribose, 0.65% glutamic acid and the balance water.

[0061] The preparation method of the reagent of this embodiment includes: weighing each component in proportion, and stirring and mixing them evenly at room temperature.

[0062] Example 2 This embodiment provides a reagent containing a composition for promoting collagen production, including the following components in mass percentage: 0.001% palmitoyl tripeptide trifluoroacetate, 0.001% methylsilanetriol, 0.001% snake venom tripeptide, 23.4% glycerol, 10.42% D-ribose, 0.65% glutamic acid and the balance water.

[0063] The preparation method of the reagent of this embodiment includes: weighing each component in proportion, and stirring and mixing them evenly at room temperature.

[0064] Example 3 This embodiment provides a reagent containing a composition for promoting collagen production, including the following components in mass percentage: 0.23% palmitoyl tripeptide trifluoroacetate, 0.005% methylsilanetriol, 0.11% snake venom tripeptide, 23.4% glycerol, 10.42% D-ribose, and 0.65% glutamic acid, and the remainder of water is added until the sum of the contents of each component is 100%.

[0065] The preparation method of the reagent of this embodiment includes: weighing each component in proportion, and stirring and mixing them evenly at room temperature.

[0066] Example 4 This embodiment provides a reagent containing a composition for promoting collagen production, including the following components in mass percentage: 0.091% palmitoyl tripeptide trifluoroacetate, 0.165% methylsilanetriol, 0.091% snake venom tripeptide, 23.4% glycerol, 10.42% D-ribose, 0.65% glutamic acid and the remainder water.

[0067] The preparation method of the reagent of this embodiment includes: weighing each component in proportion, and stirring and mixing them evenly at room temperature.

[0068] Embodiments 5 to 8 This embodiment provides a facial cream, the formula of which is shown in Table 1.

[0069] Table 1 Different cream formulas (wt%)

[0070] The preparation method of the facial cream of this embodiment comprises the following steps: (1) Heat the deionized water in phase A to 80-85°C; (2) Add the components of phase B in the order of glyceryl stearate, glyceryl stearate / PEG-100 stearate complex, isohexadecane, isooctyl palmitate, and cetearyl alcohol into an oil pan, pre-mix, heat to 80-85°C, and stir evenly; (3) Add the material obtained in step (2) to the material obtained in step (1) which has been homogenized, stir and mix evenly and homogenize to emulsify, and homogenize for 10 minutes; (4) The material obtained in step (3) is cooled to below 40°C, and the ingredients of phase C are added in the order of PE phenoxyethanol, SC50 ethylhexylglycerin, and the reagents in the example, and stirred evenly, and cooled to room temperature.

[0071] Comparative Examples 1 to 6 Comparative Examples 1 to 6 provide different reagents, and their specific compositions are shown in Table 2. The preparation method of the reagents includes: weighing each component in proportion, and stirring and mixing at room temperature until uniform.

[0072] Table 2 Composition of different compositions (wt%)

[0073] Comparative Example 7 Comparative Example 7 provides a facial cream, referring to the facial cream and preparation method thereof of Example 5, except that an equal weight of the reagent of Comparative Example 4 is used to replace the reagent of Example 1.

[0074] Comparative Example 8 Comparative Example 8 provides a facial cream, referring to the facial cream and preparation method thereof of Example 5, except that: the reagent of Example 1 is not added, and the missing amount of the reagent of Example 1 is supplemented with deionized water.

[0075] Experimental Example 1 The mitochondrial membrane potential of the reagents of Examples 1 to 4 and Comparative Examples 1 to 6 was tested. This experiment used fibroblasts as the subject to evaluate the effect of each reagent on the mitochondrial membrane potential. The specific experimental steps are as follows.

[0076] Cell inoculation: Human fibroblasts with a confluence of 80% were plated in a 12-well plate with a volume of 1 mL per well to achieve a cell density of 7 × 10 4 ~9×10 4 / hole.

[0077] Observe the cells: the cells are in good condition and the number is appropriate, so the next experiment can be carried out.

[0078] 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.

[0079] Model culture: discard the culture medium, wash with PBS, and then add CCCP (model group) for incubation.

[0080] Sample culture: Based on the model group, the culture medium was discarded and washed with PBS, and then 1 mL / well of high-glucose DMEM complete culture medium (blank group and model group) and 1 mL / well of high-glucose DMEM complete culture medium containing the composition of the embodiment / comparative example (composition group, the content of the composition in the high-glucose DMEM complete culture medium is 1 wt %) were added, and then the well plate was placed in a 37°C, 5% CO2 incubator for 24 h.

[0081] Collect cells: 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 use pre-cooled buffer to wash, centrifuge repeatedly 3 times, and discard the supernatant. Collect the cells into a new EP tube and add fresh pre-cooled buffer.

[0082] Mitochondrial fluorescence assay: 100uL of cell fluid was taken into a 96-well plate, and the fluorescence intensity of JC-1 monomer and polymer was detected respectively, and ratio analysis was performed.

[0083] Figure 1The test results of the mitochondrial membrane potential of the reagents provided in different embodiments or comparative examples of the present invention. Mitochondria in cells are the main place for aerobic respiration to produce ATP and are one of the important organelles in the body. Mitochondria use oxidizable substrates to generate an electrochemical proton gradient on the mitochondrial membrane, which is used to produce ATP. Under normal circumstances, the membrane potential of mitochondria is positive outside and negative inside. During the dysfunction of mitochondria, the mitochondrial membrane potential will decrease, which may cause the opening of the mitochondrial permeability pore and the loss of the electrochemical gradient. Therefore, the mitochondrial membrane potential is an essential parameter for 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 high, JC-1 aggregates in the matrix of mitochondria to form polymers, which can produce red fluorescence; when the mitochondrial membrane potential is low, JC-1 cannot aggregate in the matrix of mitochondria. At this time, JC-1 is a monomer and can produce green fluorescence. Therefore, the ratio of red and green fluorescence (Red / Green) is used to measure the ratio 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 ingredients have the effect of increasing mitochondrial membrane potential, improving mitochondrial function, and restoring cell health. Figure 1 It can be seen that the mitochondrial membrane potential of fibroblasts in the model group was significantly reduced, 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 which the mitochondrial membrane potential measured by 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.

[0084] Experimental Example 2 The reagents of Examples 1 to 4 and Comparative Examples 1 to 6 were tested for HSP47 protein expression. This experiment used fibroblasts as the subject to evaluate the effect of each reagent on HSP47 protein expression. The specific experimental steps are as follows.

[0085] Cell inoculation: Human fibroblasts with a confluence of 80% were plated in a 6-well plate with a volume of 2 mL per well to achieve a cell density of 20 × 10 4 / hole.

[0086] 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.

[0087] Model culture: discard the culture medium, wash with PBS, and then culture under heat stimulation conditions (model group).

[0088] Sample culture: discard the culture medium, wash with PBS, add 2 mL / well high-glucose DMEM complete culture medium (blank group and model group), 2 mL / well high-glucose DMEM complete culture medium containing the example / comparative example reagent (reagent group, the content of the reagent in the high-glucose DMEM complete culture medium is 1 wt%), then model, and then place the well plate in a 37°C, 5% CO2 incubator for 24 h.

[0089] HSP47 immunofluorescence detection: discard the culture medium, wash once or twice with PBS, perform fixation, permeabilization, and blocking processes, incubate with primary and secondary antibodies, observe under a microscope, and perform fluorescence intensity data analysis (total fluorescence intensity / number of cells).

[0090] The immunofluorescence images and fluorescence intensity comparison images of the HSP47 proteins of the reagents provided in different embodiments or comparative examples are shown in FIG. Figure 2 and Figure 3 As shown. Figure 2 , 3 It can be seen that there is no statistical difference in the HSP47 protein expression of fibroblasts in the model group and the blank group, and the HSP47 expression measured by the reagents of Examples 1 to 4 is improved, with a statistical difference of p < 0.05, among which the expression effect measured by Example 3 is better, and the HSP47 expression measured by the reagents of Comparative Examples 1 to 6 does not increase significantly.

[0091] Experimental Example 3 The reagents of Examples 1 to 4 and Comparative Examples 1 to 6 were tested for the expression of type I and type III collagen. This experiment used fibroblasts as the subject to evaluate the effect of each reagent on the expression of type I and type III collagen. The specific experimental steps are as follows.

[0092] Cell inoculation: Human fibroblasts with a confluence of 80% were plated in a 24-well plate with a volume of 2 mL per well to a cell density of 4 × 10 4 / hole.

[0093] 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.

[0094] Sample culture: discard the culture medium, wash with PBS, add high-glucose DMEM complete culture medium (blank group), high-glucose DMEM complete culture medium containing the example / comparative example reagent (reagent group, the content of the reagent in the high-glucose DMEM complete culture medium is 0.1wt%), and high-glucose DMEM complete culture medium containing the positive control (0.01wt% ascorbic acid AA) (positive control group), and then place the well plate in a 37°C, 5% CO2 incubator for 72h.

[0095] Immunofluorescence detection of type I and type III collagen: discard the culture medium, wash once or twice with PBS, perform fixation, penetration, and blocking, and then incubate type I or type III primary and secondary antibodies respectively, observe under a microscope, and perform fluorescence intensity data analysis (total fluorescence intensity / number of cells). Use Image J software to measure the total fluorescence intensity in the same area, keep the threshold consistent, and calculate the number of cells in the same area. The average fluorescence intensity = total fluorescence intensity / number of cells. The final result is based on the blank group as 100%, and the relative content of each group is calculated.

[0096] The comparison charts of the expression results of type I collagen and type III collagen of the reagents provided in different embodiments or comparative examples of the present invention are shown in FIG. Figure 4 and Figure 5 As shown. Figure 4 , 5 It can be seen that the reagents of the positive control group and Examples 1 to 4 all enhanced the content of type I and type III collagen, with a statistical difference of p < 0.05, among which Example 3 had a better implementation effect; the reagents of Comparative Examples 1 to 6 had no significant effect on collagen enhancement.

[0097] Experimental Example 4 The facial creams of Examples 5 to 8 and Comparative Examples 7 to 8 were subjected to a volunteer usage experiment.

[0098] Experimental steps: recruit several volunteers aged between 30 and 60 with mild or severe facial wrinkles and sagging skin, collect basic data (depth of nasolabial folds and skin elasticity) and then perform radiofrequency treatment. Randomly select 3 volunteers to use the cream of comparative example 8 (placebo), 3 volunteers to use the cream of comparative example 7, 3 volunteers to use the cream of example 5, 3 volunteers to use the cream of example 6, 3 volunteers to use the cream of example 7, and 3 volunteers to use the cream of example 8. Use the cream once in the morning and once in the evening every day. Return visits and collect the above skin data in the 8th week. The test results are shown in Table 3, where: The difference between wrinkle depth and basic value = wrinkle depth (basic value) - wrinkle depth (after use); The change rate of skin elasticity relative to the basic value % = [skin elasticity (after use) - skin elasticity (basic value)] / skin elasticity (basic value) × 100%.

[0099] Table 3 Changes in wrinkle depth and skin elasticity among users of different facial creams

[0100] Table 3 shows that the results of Example 7 are that the difference in wrinkle depth relative to the basic value and the rate of change of skin elasticity relative to the basic value are the largest, indicating that the anti-wrinkle effect implemented on the skin is the best.

[0101] Figure 6 The following is a comparison chart of the changes in wrinkle depth before and after volunteers used the creams prepared in different embodiments or comparative examples of the present invention. From the above test results, it can be seen that compared with the placebo cream and the basic value of comparative example 8, after using the cream of the embodiment for 8 weeks, the average depth of skin texture was significantly reduced, the skin elasticity was significantly increased, and the improvement effect of embodiment 7 on wrinkle depth and elasticity was the best (the green shaded part of the image was the smallest. Image after 8 weeks). There was no significant difference before and after using comparative example 7.

[0102] In summary, the composition or reagent of the present invention can promote the production of type I and type III collagen by promoting the expression of HSP47 protein. In human tests, it was found that after using a facial cream containing the corresponding reagent for 8 weeks, the depth of nasolabial folds was significantly reduced and skin elasticity was significantly increased, which once again demonstrated that the composition or reagent of the present invention can promote collagen production.

[0103] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A composition, characterized in that The composition comprises, by weight, 0.001 to 0.55 parts of palmitoyl tripeptide trifluoroacetate, 0.001 to 1 part of methylsilanetriol and 0.001 to 0.55 parts of snake venom-like tripeptide.

2. The composition according to claim 1, characterized in that In the composition, the mass ratio of palmitoyl tripeptide trifluoroacetate, methylsilanetriol and snake venom-like tripeptide is (30-60) : 1 : (10-40).

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

4. A reagent, characterized in that The reagent contains the composition according to any one of claims 1 to 3, and the mass content of the composition in the reagent is 0.003% to 2.1% based on the total mass of the reagent.

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

6. The reagent according to claim 5, characterized in that The reagent comprises the following components in percentage by mass: 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.

7. Use of a composition according to any one of claims 1 to 3 or an agent according to any one of claims 4 to 6 in the preparation of a preparation having at least one of the following characteristics: (a) maintain and / or increase mitochondrial activity; (b) Enhanced HSP47 protein expression; (c) Enhanced the expression of type I and type III collagen.

8. A type I collagen and type III collagen production promoter, characterized in that: The accelerator comprises the composition according to any one of claims 1 to 3 or the reagent according to any one of claims 4 to 6.

9. A cosmetic, characterized in that: The cosmetic comprises the composition according to any one of claims 1 to 3 or the reagent according to any one of claims 4 to 6 and cosmetically acceptable auxiliary materials.

10. The cosmetic according to claim 9, characterized in that: The mass percentage of the composition or reagent is no more than 5% based on the total mass of the cosmetic.

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