Nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects, preparation method and application thereof

Abstract

The present application discloses a nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects, and its preparation method and application, which belongs to the field of cosmetic technology. The present application prepares a nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects by nano-encapsulating liposome bilayer membrane flow mosaic regulator, bilayer film-forming agent, active ingredient solvent, pterostilbene, cycloastragenol, baicalein, quercetin, soluble phosphopeptide, salvianolic acid B, and glycerol, and at the same time utilizes the property that salvianolic acid B spontaneously forms a hydrogel when combined with a soluble phosphopeptide aqueous solution. The present application provides a theoretical and experimental basis for the reasonable combination of pterostilbene and three traditional Chinese medicine active substances, cycloastragenol, baicalein, and quercetin, in terms of functional ingredients, and has application prospects in the production of whitening, anti-oxidation, anti-inflammatory and anti-aging cosmetics.

Description

Technical Field

[0001] The present application belongs to the technical field of cosmetics, and in particular relates to a nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects, and a preparation method and application thereof. Background Art

[0002] External stimuli (such as drugs, ultraviolet rays and ionizing radiation), endogenous free radicals and reactive oxygen species (ROS) can directly or indirectly damage cellular components such as proteins, lipids and DNA, causing damage to human skin, accelerating skin aging, wrinkles and dullness. Inflammation is often accompanied by oxidative stress. Excessive free radicals in the human body lead to an imbalance of inflammatory homeostasis, which accelerates the aging process, and aging causes the metabolic balance of free radicals to be disrupted. In order to resist these adverse effects, the body has formed a complex oxidative stress response system to alleviate the damage to cells, such as the regulation of the nuclear factor E2-related factor (Nrf2) signaling pathway and the nuclear factor κB (NF-κB) signaling pathway. Nrf2 is a key cytokine in the anti-oxidative stress response. After effectively binding to the antioxidant response element (ARE), it can activate downstream antioxidant genes: including superoxide dismutase (SOD2, SOD1), heme oxygenase (HO-1), quinone oxidoreductase (NQO1), and catalase to fight oxidative stress. NF-κB is one of the important transcription factors that regulate inflammatory responses. The NF-κB signaling pathway can inhibit the secretion of SASP, the active molecule of senescent cells, alleviate the inflammatory state caused by the continuous secretion of SASP, and resist the occurrence of the hallmark pathological changes of aging.

[0003] The prior art with application number CN2021107649291 discloses a method for preparing a skin anti-aging composition, which consists of a cysteine ​​derivative, ribose, vitamin E, carnosine, phosphatidylglycerol, poloxamer 407, a polyol, and the balance is deionized water. The prior art with application number CN2022112099366 discloses a multi-layer moisturizing, repairing, wrinkle-removing and anti-aging peony cream and a preparation method thereof. The peony cream is composed of peony seed oil, shea butter, jojoba seed oil, olive oil, vaseline, vitamin E, butylene glycol, polyacrylate cross-linked polymer-6, medium and small molecule sodium hyaluronate, bosin, defuxiu, astaxanthin, essence, cyclopolydimethylsiloxane and polydimethylsiloxane cross-linked polymer, self-emulsifying monoglyceride, 16-18 alcohol, silicone elastomer gel, ethylhexylglycerin, caprylhydroxamic acid, glyceryl caprylate, nano-encapsulated retinol retinoic acid ester, snake venom peptide, tripeptide-1-tetrapeptide-7, Cu peptide and water.

[0004] In summary, the anti-aging raw materials and skin care products in the prior art generally use a single antioxidant or anti-inflammatory substance, or use low-content plant extracts, or are unable to remove free radicals to solve the problem of skin aging from the root. Therefore, it is necessary to develop cosmetics that can achieve anti-inflammatory repair, improve targeting, stability and other functions from the root. Summary of the invention

[0005] The present application discloses a nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects, and a preparation method and application thereof, thereby solving the technical problem of skin aging from the root, that most anti-aging raw materials and skin care products in the technology use a single substance with antioxidant or anti-inflammatory effects, or use low-content plant extracts, or are unable to remove free radicals.

[0006] In order to achieve the above-mentioned purpose, the first aspect of the present application provides a nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects. The nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects of the present application is composed of the following components in mass percentage:

[0007] 5-20wt% liposome bilayer membrane flow mosaic regulator, 1-10wt% bilayer film-forming agent, 5-20wt% active ingredient solvent, 0.1-5wt% pterostilbene, 0.1-5wt% cycloastragenol, 0.1-5wt% baicalein, 0.1-5wt% quercetin, 1-10wt% soluble phosphopeptide, 1-5wt% salvianolic acid B, 10-30wt% glycerol, and the balance is ultrapure water.

[0008] In some embodiments, the liposome bilayer membrane fluid mosaic modulator is at least one of cholesterol, 1,3-propylene glycol, 1,2-pentanediol, 1,2-hexanediol, chitosan, or yeast β-glucan.

[0009] In some embodiments, the bilayer film former is at least one of lecithin, soy lecithin, hydrogenated lecithin, or egg yolk lecithin.

[0010] In some embodiments, the active ingredient solvent is at least one of PEG-8 caprylic triglyceride, PEG-8 capric triglyceride, PEG-40 hydrogenated castor oil, diisopropyl sebacate, polysorbate-20, or polyglyceryl-10 stearate.

[0011] In some embodiments, the pterostilbene, cycloastragenol, baicalin, quercetin, soluble phosphopeptide, and salvianolic acid B are of natural origin, biosynthetic origin, or chemical synthetic origin.

[0012] The second aspect of the present application provides a method for preparing a nanoemulsion gel, comprising the following steps:

[0013] S1: After the active ingredient solvent and glycerol are mixed to form a mixed solution, pterostilbene, cycloastragenol, baicalein and quercetin are ultrasonically dispersed in the mixed solution to obtain phase A;

[0014] S2: After mixing the liposome bilayer membrane flow mosaic regulator, bilayer membrane-forming agent, soluble phosphopeptide and ultrapure water, the mixture is stirred under heating in a water bath to obtain phase B;

[0015] S3: under shearing conditions, adding phase A to phase B for mixing, homogenizing and shearing to obtain liposome colostrum;

[0016] S4: subjecting the liposome colostrum to ultra-high pressure homogenization and cooling it to room temperature to obtain a liposome emulsion;

[0017] S5: At room temperature, salvianolic acid B is added to the liposome emulsion, and the solution is solubilized with ultrasonic aid and then incubated overnight to obtain a nanoemulsion gel.

[0018] In some embodiments, in step S2, the water bath heating temperature is 40-50°C, and the stirring rate is 200-500 rpm.

[0019] In some embodiments, in step S3, the shearing speed is 5000-8000 rpm, the shearing temperature is 40-50° C., and the shearing time is 5-10 min.

[0020] In some embodiments, in step S4, the pressure is 500-1000 bar, the temperature is 40-80°C, and the cycle is 6-9 times; in step S5, the ultrasonic power is 80-100w, the dissolution time is 30-40min, and the incubation temperature is 4°C.

[0021] The third aspect of the present application provides the use of the nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects described in the present application in cosmetics.

[0022] Compared with the prior art, the advantages or beneficial effects of the embodiments of the present application include at least:

[0023] The nanoemulsion gel provided in the first aspect of the present application, on the one hand, can cooperate and jointly regulate in two important anti-aging signaling pathways, Nrf2 and NF-κB signaling pathways, through the compounding of pterostilbene, cycloastragenol, baicalin and quercetin in specific proportions, so as to more comprehensively enhance the antioxidant capacity of cells and reduce oxidative stress damage, so as to achieve the purpose of synergistic enhancement. Pterostilbene, cycloastragenol, baicalin, and quercetin have some overlap in anti-aging mechanisms and effects, but they are also different; on the other hand, the regulatory effects of pterostilbene and other components of the present application can complement each other, and the antioxidant and anti-inflammatory effects are better than those of single extracts or other compound compositions, which can repair inflammation from the root, reduce fine lines, and improve skin elasticity; on the third hand, the liposome bilayer membrane flow mosaic regulator in the present application method can provide good rheological properties and ion resistance, improve the stability of the liposome emulsion, and effectively increase the stability of the pterostilbene, cycloastragenol, baicalin, and quercetin nano-encapsulation system during the storage period and the efficacy and activity during application; on the fourth hand, the present application adopts a hydrogel scaffold, which can prolong the administration time, form a drug reservoir in the dermis, and improve bioavailability. Hydrogels are highly biocompatible in therapeutic applications and can provide a shielding effect on the harmful effects of active substances directly acting on the skin. Hydrogels have strong antioxidant activity, can eliminate excess ROS around the wound, and help wound healing. Liposomes containing pterostilbene, cycloastragenol, baicalein and quercetin are loaded into a hydrogel network, and the resulting nanoemulsion gel has excellent rheological properties and strong adhesion, improves carrier stability and skin retention time, and has efficient self-healing properties and antioxidant capacity. In addition, in the present application, salvianolic acid B (SAB) spontaneously forms a hydrogel when combined with an aqueous solution of a soluble phosphopeptide (NapFFKYp, molecule 1), and the pH of the gel is about 6.0, which is very similar to the pH of normal human skin.

[0024] The preparation method of the nanoemulsion gel provided in the second aspect of the present application, on the one hand, the nano-encapsulation system prepared by the method of the present application can protect the active ingredients from the influence of the external environment, control the release rate, enhance the efficacy of the drug, and make the drug show a satisfactory therapeutic effect. The natural extract itself has many advantages such as low drug toxicity and side effects and good tolerance. The four natural compounds with synergistic effects are combined and prepared into a nano-encapsulation system, which can improve its bioavailability, stability and targeting. On the other hand, the ultrasonic dispersion technology is used in the method of the present application to disperse the active substances pterostilbene, cycloastragenol, baicalein, and quercetin to improve the dispersibility of the active substances in the medium. Ultrasonic dispersion uses the cavitation effect generated by ultrasound to achieve the dispersion of solid particles in the liquid. The cavitation effect can produce a high temperature and high pressure environment, which is conducive to the formation of nanoparticles. Ultrasonic dispersion can also reduce the binding force between particles and effectively prevent agglomeration. On the third aspect, the mixed homogenization shearing in the method of the present application can reduce the particle size of the dispersed phase in the material, obtain finer dispersed particles, and improve the uniformity and stability of the material. Ultra-high pressure homogenization can produce very fine and uniform particles with higher pressure than traditional high pressure homogenizers, which can provide better stability, improve storage quality, improve homogeneity, improve absorption quality, save expensive additives, change viscosity and reduce reaction time.

[0025] The use of the nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects in cosmetics provided in the third aspect of the present application has application prospects in the production of whitening, anti-oxidation, anti-inflammatory and anti-aging cosmetics. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments recorded in the present application, and for ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0027] Figure 1 The results of the effects of the nanoemulsion gels prepared in Examples 1-4 and Comparative Examples 1-3 of the present application on the expression of inflammatory factors TNF-α and IL-6 in RAW264.7 cells are shown in the figure; among them, the comparison between the LPS group and the blank group is ####P<0.0001; compared with the LPS group, *P<0.05, **P<0.01, ***P<0.005, ****P<0.0001;

[0028] Figure 2The graph shows the effect of the nanoemulsion gel prepared in Examples 1-4 and Comparative Examples 1-3 of the present application on improving H2O2 oxidative damage in HaCaT cells; wherein, the H2O2 group is compared with the blank group ####P<0.0001; compared with the H2O2 group *P<0.05, **P<0.01, ***P<0.005, ****P<0.0001;

[0029] Figure 3 The graph is a result of the nanoemulsion gel prepared in Examples 1-4 and Comparative Examples 1-3 of the present application on improving UV damage to HDF cells; wherein, the UV group is compared with the blank group ####P<0.0001; compared with the UV group *P<0.05, **P<0.01, ***P<0.005, ****P<0.0001;

[0030] Figure 4 This is a graph showing the results of the chicken embryo test of the nanoemulsion gel prepared in Examples 1-4 of the present application. DETAILED DESCRIPTION

[0031] The technical solutions in the embodiments of the present application will be described clearly and completely below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, rather than all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0032] In the following description of this embodiment, the term "and / or" is used to describe the association relationship of associated objects, indicating that there may be three relationships. For example, A and / or B can represent: A exists alone, B exists alone, and A and B exist at the same time. A and B can be singular or plural. The character " / " generally indicates that the associated objects before and after are in an "or" relationship.

[0033] In the following description of this embodiment, the term "at least one" refers to one or more, and "plurality" refers to two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single items or plural items. For example, "at least one of a, b or c", or "at least one of a, b and c", can all represent: a, b, c, ab (i.e., a and b), ac, bc, or abc, where a, b, c can be single or multiple, respectively.

[0034] The terms used in the embodiments of the present application are only for the purpose of describing specific embodiments, and are not intended to limit the present application. The singular forms "a", "an" and "the" used in the embodiments of the present application and the appended claims are also intended to include plural forms, unless the context clearly indicates other meanings.

[0035] Those skilled in the art should understand that in the following description of the embodiments of the present application, the order of serial numbers does not mean the order of execution, some or all of the steps can be executed in parallel or sequentially, and the execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0036] Those skilled in the art will appreciate that the numerical ranges in the embodiments of the present application are to be understood as each intermediate value between the upper and lower limits of the scope also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present application. The upper and lower limits of these smaller ranges may be independently included or excluded in the scope.

[0037] Unless otherwise specified, the technical / scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which this application belongs. Although this application only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of this application. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0038] The first aspect of the present application provides a nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects. The nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects of the present application is composed of the following components in mass percentage:

[0039] 5-20wt% liposome bilayer membrane flow mosaic regulator, 1-10wt% bilayer film-forming agent, 5-20wt% active ingredient solvent, 0.1-5wt% pterostilbene, 0.1-5wt% cycloastragenol, 0.1-5wt% baicalein, 0.1-5wt% quercetin, 1-10wt% soluble phosphopeptide, 1-5wt% salvianolic acid B, 10-30wt% glycerol, and the balance is ultrapure water.

[0040] In the embodiments of the present application, the liposome bilayer membrane fluid mosaic regulator is at least one of cholesterol, 1,3-propylene glycol, 1,2-pentanediol, 1,2-hexanediol, chitosan or yeast β-glucan.

[0041] In the embodiment of the present application, the bilayer film-forming agent is at least one of lecithin, soybean lecithin, hydrogenated lecithin or egg yolk lecithin.

[0042] In the embodiments of the present application, the active ingredient solvent is at least one of PEG-8 caprylic triglyceride, PEG-8 capric triglyceride, PEG-40 hydrogenated castor oil, diisopropyl sebacate, polysorbate-20 or polyglyceryl-10 stearate.

[0043] In the examples of the present application, the pterostilbene, cycloastragenol, baicalin, quercetin, soluble phosphopeptide, and salvianolic acid B are of natural origin, biosynthetic origin, or chemical synthetic origin.

[0044] In the embodiment of the present application, the liposome bilayer membrane flow mosaic regulator is preferably cholesterol and deacetylated chitosan. The bilayer film-forming agent is preferably egg yolk lecithin. The pterostilbene, cycloastragenol, baicalein, quercetin, soluble phosphopeptide, and salvianolic acid B are preferably derived from chemical synthesis.

[0045] The second aspect of the present application provides a method for preparing a nanoemulsion gel having anti-aging and mitochondrial telomerase activation effects, comprising the following steps:

[0046] S1: After the active ingredient solvent and glycerol are mixed to form a mixed solution, pterostilbene, cycloastragenol, baicalein and quercetin are ultrasonically dispersed in the mixed solution to obtain phase A;

[0047] S2: After mixing the liposome bilayer membrane flow mosaic regulator, bilayer membrane-forming agent, soluble phosphopeptide and ultrapure water, stirring under heating conditions of 40-50° C. in a water bath at a stirring rate of 200-500 rpm, phase B is obtained;

[0048] S3: under shearing conditions, adding phase A into phase B for mixing, homogenizing and shearing, with a shearing speed of 5000-8000 rpm, a shearing temperature of 40-50° C., and a shearing time of 5-10 min to obtain liposome colostrum;

[0049] S4: subjecting the liposome colostrum to ultra-high pressure homogenization treatment at a pressure of 500 to 1000 bar and a temperature of 60 to 80° C. for 6 to 9 cycles, and cooling to room temperature to obtain a liposome emulsion;

[0050] S5: under normal temperature, add salvianolic acid B to the liposome emulsion, dissolve it with ultrasound at 80-100w for 30-40min, and then incubate it at 4°C overnight to obtain a nanoemulsion gel.

[0051] The third aspect of the present application provides the use of the nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects described in the present application in cosmetics.

[0052] The first aspect of the present application provides a nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects. On the one hand, a composition comprising pterostilbene, cycloastragenol, baicalein, and quercetin is subjected to nano-encapsulation treatment, and the characteristic of spontaneously forming a hydrogel when salvianolic acid B (SAB) and a soluble phosphopeptide (NapFFKYp, molecule 1) aqueous solution are used to prepare a nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects. The theoretical and experimental basis is provided for the reasonable combination of pterostilbene and three traditional Chinese medicine active substances cycloastragenol, baicalein, and quercetin in terms of functional ingredients, and it has application prospects in the production of whitening, anti-oxidation, anti-inflammatory and anti-aging cosmetics. On the other hand, pterostilbene, baicalein, and quercetin induce activation of Nrf2 protein, and the increase in the expression of Nrf2 in cells can reduce the expression of NF-κB protein in the NF-κB signal transduction pathway. At the same time, since cycloastragenol inhibits the NF-κB signaling pathway, pterostilbene, baicalein, and quercetin can enhance this inhibitory effect, thereby inhibiting NF-κB transcriptional activity and cell apoptosis trends. Cycloastragenol activates telomerase targets, inhibits the production of inflammatory factor interleukin-1β, and enhances the effect of inflammatory inhibition. Baicalein and quercetin can synergistically induce Nrf2 activation, bind to antioxidant response elements (ARE), enhance the expression of Nrf2 downstream genes GCLC, GCLM, and NQO1, enhance the antioxidant capacity of cells, and reduce hydrogen peroxide-induced oxidative stress damage. Baicalein can inhibit the release of inflammatory cytokines, and quercetin can induce the expression or activation of HO-1, inhibiting oxidative stress response. The two can more comprehensively regulate the oxidative stress pathway through the complementary regulatory mechanisms. Thirdly, the liposome bilayer membrane flow mosaic regulator can provide good rheological properties and ion resistance, improve the stability of the liposome emulsion, and effectively increase the stability of the pterostilbene, cycloastragenol, baicalin, and quercetin nano-encapsulation system during the storage period and the efficacy and activity during application.

[0053] The second aspect of the present application provides a method for preparing a nanoemulsion gel. On the one hand, the active substances pterostilbene, cycloastragenol, baicalein, and quercetin are dispersed using ultrasonic dispersion technology to improve the dispersibility of the active substances in the medium. Ultrasonic dispersion uses the cavitation effect generated by ultrasound to achieve the dispersion of solid particles in a liquid. The cavitation effect can produce a high temperature and high pressure environment, which is conducive to the formation of nanoparticles. Ultrasonic dispersion can also reduce the binding force between particles and effectively prevent agglomeration. On the other hand, the mixed homogenization shearing in the present application method can reduce the particle size of the dispersed phase in the material, obtain finer dispersed particles, and improve the uniformity and stability of the material. Ultrahigh pressure homogenization can prepare very fine and uniform particles, which has a higher pressure than traditional high pressure homogenizers, can provide better stability, improve storage quality, improve homogeneity, improve absorption quality, save expensive additives, change viscosity, and reduce reaction time.

[0054] The technical solution of the present application will be further described below in conjunction with specific embodiments.

[0055] Example 1

[0056] A nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects, composed of the following components in percentage by mass:

[0057] 3% pterostilbene, 2% cycloastragenol, 2% baicalein, 2% quercetin, 5% PEG-8 caprylic triglyceride, 5% PEG-40 hydrogenated castor oil, 20% glycerol, 5% hydrogenated lecithin, 2% egg yolk lecithin, 1% cholesterol, 4% 1,3-propylene glycol, 3% deacetylated chitosan, 10% soluble phosphopeptide, 4% salvianolic acid B, 32% ultrapure water.

[0058] The pterostilbene, cycloastragenol, baicalin, quercetin, soluble phosphopeptide and salvianolic acid B are derived from chemical synthesis.

[0059] The method for preparing the nanoemulsion gel having anti-aging and mitochondrial telomerase activation effects comprises the following steps:

[0060] S1: Weigh each component according to the mass percentage, mix PEG-8 caprylic triglyceride, PEG-40 hydrogenated castor oil and glycerin to form a mixed solution, disperse pterostilbene, cycloastragenol, baicalein and quercetin in the mixed solution by ultrasonication at room temperature for 30 min to obtain phase A;

[0061] S2: hydrogenated lecithin, egg yolk lecithin, cholesterol, 1,3-propylene glycol, chitosan, soluble phosphopeptide and ultrapure water were mixed, heated in a water bath at 45°C, stirred with a magnetic stirrer at 400 rpm for 40 min, and kept warm in a water bath to obtain phase B;

[0062] S3: Under shearing conditions, phase A is added to phase B for mixing, homogenization and shearing, with a shearing speed of 8000 rpm, a shearing temperature of 50°C and a shearing time of 7 min to obtain liposome colostrum;

[0063] S4: subjecting the liposome colostrum to ultra-high pressure homogenization treatment at a pressure of 800 bar and a temperature of 70° C. for 8 cycles, and cooling to room temperature to obtain a liposome emulsion;

[0064] S5: At room temperature, salvianolic acid B was added to the liposome emulsion, solubilized by ultrasound at 100W for 30min, and then incubated at 4°C overnight to obtain a nanoemulsion gel.

[0065] Example 2

[0066] A nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects, composed of the following components in percentage by mass:

[0067] 3% pterostilbene, 2% cycloastragenol, 2% baicalein, 2% quercetin, 5% polyglyceryl-10 stearate, 5% polysorbate-20, 20% glycerol, 5% soy lecithin, 2% hydrogenated lecithin, 1% chitosan, 4% 1,2-pentanediol, 3% 1,2-hexanediol, 10% soluble phosphopeptide, 4% salvianolic acid B, 32% ultrapure water.

[0068] The pterostilbene, cycloastragenol, baicalin, quercetin, soluble phosphopeptide and salvianolic acid B are derived from chemical synthesis.

[0069] The method for preparing the nanoemulsion gel having anti-aging and mitochondrial telomerase activation effects comprises the following steps:

[0070] S1: Weigh each component according to the mass percentage, mix polyglycerol-10 stearate, polysorbate-20 and glycerol to form a mixed solution, disperse pterostilbene, cycloastragenol, baicalein and quercetin in the mixed solution by ultrasonication at room temperature for 25 min to obtain phase A;

[0071] S2: Soy lecithin, hydrogenated lecithin, chitosan, 1,2-pentanediol, 1,2-hexanediol, soluble phosphopeptide and ultrapure water were mixed, heated in a 50°C water bath, stirred with a magnetic stirrer at 200 rpm for 30 min, and kept warm in a water bath to obtain phase B;

[0072] S3: Under shearing conditions, phase A is added to phase B for mixing, homogenization and shearing, with a shearing speed of 6000 rpm, a shearing temperature of 40°C and a shearing time of 10 min to obtain liposome colostrum;

[0073] S4: subjecting the liposome colostrum to ultra-high pressure homogenization treatment at a pressure of 600 bar and a temperature of 60° C. for 9 cycles, and cooling to room temperature to obtain a liposome emulsion;

[0074] S5: under normal temperature conditions, salvianolic acid B was added to the liposome emulsion, solubilized by ultrasound at 90W for 40min, and then incubated at 4°C overnight to obtain a nanoemulsion gel.

[0075] Example 3

[0076] A nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects, composed of the following components in percentage by mass:

[0077] 5% pterostilbene, 0.1% cycloastragenol, 5% baicalein, 0.1% quercetin, 5% PEG-8 capric triglyceride, 30% glycerol, 10% egg yolk lecithin, 20% yeast β-glucan, 1% soluble phosphopeptide, 1% salvianolic acid B, 22.8% ultrapure water.

[0078] The pterostilbene, cycloastragenol, baicalin, quercetin, soluble phosphopeptide and salvianolic acid B are of natural origin.

[0079] The method for preparing the nanoemulsion gel having anti-aging and mitochondrial telomerase activation effects comprises the following steps:

[0080] S1: Weigh each component according to the mass percentage, mix PEG-8 capric triglyceride and glycerol to form a mixed solution, disperse pterostilbene, cycloastragenol, baicalein and quercetin in the mixed solution by ultrasonication at room temperature for 25 min to obtain phase A;

[0081] S2: egg yolk lecithin, yeast β-glucan, soluble phosphopeptide and ultrapure water were mixed, heated in a water bath at 40°C, stirred with a magnetic stirrer at 500 rpm for 60 min, and kept warm in a water bath to obtain phase B;

[0082] S3: Under shearing conditions, phase A is added to phase B for mixing, homogenization and shearing, with a shearing speed of 5000 rpm, a shearing temperature of 40°C and a shearing time of 5 min to obtain liposome colostrum;

[0083] S4: subjecting the liposome colostrum to ultra-high pressure homogenization treatment at a pressure of 500 bar and a temperature of 80° C. for 7 cycles, and cooling to room temperature to obtain a liposome emulsion;

[0084] S5: under normal temperature conditions, salvianolic acid B was added to the liposome emulsion, solubilized by 80W ultrasound for 40 min, and then incubated at 4°C overnight to obtain a nanoemulsion gel.

[0085] Example 4

[0086] A nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects, composed of the following components in percentage by mass:

[0087] 0.1% pterostilbene, 5% cycloastragenol, 0.1% baicalein, 5% quercetin, 20% diisopropyl sebacate, 10% glycerol, 1% lecithin, 5% chitosan, 5% soluble phosphopeptide, 5% salvianolic acid B, 43.8% ultrapure water.

[0088] The pterostilbene, cycloastragenol, baicalin, quercetin, soluble phosphopeptide and salvianolic acid B are biosynthetic sources.

[0089] The method for preparing the nanoemulsion gel having anti-aging and mitochondrial telomerase activation effects comprises the following steps:

[0090] S1: Weigh the components according to the mass percentage, mix diisopropyl sebacate and glycerol to form a mixed solution, disperse pterostilbene, cycloastragenol, baicalein and quercetin in the mixed solution by ultrasonication at room temperature for 25 min to obtain phase A;

[0091] S2: Mix lecithin, chitosan, soluble phosphopeptide and ultrapure water, heat in a water bath at 40°C, stir with a magnetic stirrer at 500 rpm for 60 min, and keep warm in a water bath to obtain phase B;

[0092] S3: Under shearing conditions, phase A is added to phase B for mixing, homogenization and shearing, with a shearing speed of 5000 rpm, a shearing temperature of 40°C and a shearing time of 5 min to obtain liposome colostrum;

[0093] S4: subjecting the liposome colostrum to ultra-high pressure homogenization treatment at a pressure of 1000 bar and a temperature of 40° C. for 6 cycles, and cooling to room temperature to obtain a liposome emulsion;

[0094] S5: under normal temperature conditions, salvianolic acid B was added to the liposome emulsion, solubilized by 80W ultrasound for 40 min, and then incubated at 4°C overnight to obtain a nanoemulsion gel.

[0095] Comparative Example 1: Nanoemulsion gel without quercetin

[0096] A nanoemulsion gel without quercetin, consisting of the following components in percentage by mass:

[0097] 3% pterostilbene, 3% cycloastragenol, 3% baicalein, 5% PEG-8 caprylic triglyceride, 5% PEG-40 hydrogenated castor oil, 20% glycerol, 5% hydrogenated lecithin, 2% egg yolk lecithin, 1% cholesterol, 4% 1,3-propylene glycol, 3% deacetylated chitosan, 10% soluble phosphopeptide, 4% salvianolic acid B, 32% ultrapure water.

[0098] The preparation method and parameters of the nanoemulsion gel without quercetin are the same as those in Example 1.

[0099] Comparative Example 2: Nanoemulsion gel without baicalin and quercetin

[0100] A nanoemulsion gel without baicalein and quercetin, comprising the following components in percentage by mass:

[0101] 5% pterostilbene, 4% cycloastragenol, 5% PEG-8 caprylic triglyceride, 5% PEG-40 hydrogenated castor oil, 20% glycerol, 5% hydrogenated lecithin, 2% egg yolk lecithin, 1% cholesterol, 4% 1,3-propylene glycol, 3% deacetylated chitosan, 10% soluble phosphopeptide, 4% salvianolic acid B, 32% ultrapure water.

[0102] The preparation method and parameters of the nanoemulsion gel without baicalein and quercetin are the same as those in Example 1.

[0103] Comparative Example 3: Nanoemulsion gel without cycloastragenol, baicalein and quercetin

[0104] A nanoemulsion gel without cycloastragenol, baicalein and quercetin, comprising the following components in percentage by mass:

[0105] 9% pterostilbene, 5% PEG-8 caprylic triglyceride, 5% PEG-40 hydrogenated castor oil, 20% glycerol, 5% hydrogenated lecithin, 2% egg yolk lecithin, 1% cholesterol, 4% 1,3-propylene glycol, 3% chitosan, 10% soluble phosphopeptide, 4% salvianolic acid B, 32% ultrapure water.

[0106] The preparation method and parameters of the nanoemulsion gel without cycloastragenol, baicalein and quercetin are the same as those in Example 1.

[0107] The application performance test of the nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects in cosmetics described in this application is as follows:

[0108] Test Example 1: Stability Study

[0109] The nanoemulsion gels with anti-aging and mitochondrial telomerase activation effects prepared in Examples 1-4 and the nanoemulsion gels prepared in Comparative Examples 1-3 were placed in sealed containers, respectively, and placed at -18°C, 4°C, room temperature, and 45°C for 180 days. The appearance of the samples was checked at room temperature for precipitation or discoloration in the initial state, 90 days, and 180 days, and the active substance retention rate and particle size of the samples were tested. The results are shown in Table 1.

[0110] Table 1 Stability test results of nanoemulsion gel

[0111]

[0112]

[0113]

[0114] The ability of cosmetics to maintain their initial properties or properties that do not change over time under specified or reasonably foreseeable storage and use conditions. The test results in Table 1 show that the nanoemulsion gels with anti-aging and mitochondrial telomerase activation effects prepared in Examples 1-4 and the nanoemulsion gels prepared in Comparative Examples 1-3 are stable after being dispersed in water, without agglomeration or stratification, and the nanoemulsion gels prepared in Examples 1-4 have a smaller change range than the nanoemulsion gels prepared in Comparative Examples 1-3, and the system is more stable, which is the best experimental effect.

[0115] Test Example 2: Test on the effect on the expression of inflammatory factors

[0116] RAW264.7 cells in the logarithmic growth phase were inoculated in a 24-well plate with a cell density of (2-5)×105 cells / mL, 0.5 mL per well, and incubated for 24 hours. After 24 hours, the supernatant was aspirated and 0.5 mL of the following samples were added respectively: ① Solvent control group: DMEM culture medium (2% FBS; 1% double antibody) acted on the cells for 24 hours; ② LPS inflammation model group: 1 μg / mL of LPS continuously stimulated RAW 264.7 cells for 24 hours; ③ Test group: Example 1, Example 2, Example 3, Example 4, Comparative Example 1, Comparative Example 2, Comparative Example 3 (1%) + LPS (1 μg / ml) acted on the cells together for 24 hours. After the sample treated the cells for 24 hours, the supernatant was aspirated into a centrifuge tube with a pipette, centrifuged at 1000 r / min for 5 minutes, the precipitate was discarded and the supernatant was aspirated, and the changes in the inflammatory factors in the supernatant were detected using TNF-α and IL-6 inflammatory factor detection kits. The detection method strictly followed the instructions on the detection kit. Results are shown in Figure 1 .

[0117] Depend on Figure 1 It can be seen that the levels of TNF-α and IL-6 in the LPS group were significantly higher than those in the normal group, indicating that the inflammatory induction model was successfully established. Examples 1-4 and Comparative Examples 1-2 can cause a significant decrease in the level of inflammatory factor TNF-α in RAW 264.7 cells, and the difference is statistically significant (P < 0.01), and the effects of Example 1, Example 2, Example 3, and Example 4 in inhibiting the inflammatory factor TNF-α are better than those in the comparative example. Examples 1-4 and Comparative Example 1 can cause a significant decrease in the level of inflammatory factor IL-6 in RAW 264.7 cells, and the difference is statistically significant (P < 0.01), and the effects of Example 1, Example 2, Example 3, and Example 4 in inhibiting the inflammatory factor IL-6 are better than those in the comparative example. The results show that the nanoemulsion gel samples prepared in Examples 1-4 can significantly reduce the expression of cellular inflammatory factors, and the effects are stronger than those in Comparative Examples 1-3. Among them, the nanoemulsion gel with anti-aging and mitochondrial telomerase activation prepared in Example 1 has the strongest effect on reducing the expression of inflammatory factors.

[0118] Test Example 3: Antioxidant effect test on human epidermal cells

[0119] Dilute Examples 1-4 and Comparative Examples 1-3 with DMEM medium at a final concentration of 10 μg / ml of active ingredients, respectively, and dilute hydrogen peroxide to 800 μM with DMEM medium. HaCaT cells in the logarithmic growth phase were inoculated in a 96-well plate at 8×103 per well, with a system of 100 μL per well. The outermost circle of the 96-well plate was sealed with 100 μL of PBS. After the cells were completely attached to the wall, they were replaced with complete medium containing the sample to be tested. After continuing to culture the cells for 24 hours, the CCK-8 method was used to detect cell activity. After that, a hydrogen peroxide dilution was given, and the model was cultured in an incubator for 4 hours. Immediately after 4 hours, the cell activity was detected using the CCK8 method. The results are shown in Figure 2 .

[0120] Depend on Figure 2 The results show that after hydrogen peroxide injury, the cell viability of the non-administered group was 41.3%, indicating that the hydrogen peroxide injury HaCaT model was successfully established. Compared with the blank control, the nanoemulsion gels prepared in Examples 1-4 and Comparative Example 1 can significantly improve the viability of oxidatively damaged human epidermal cells, and the difference is statistically significant (P < 0.01), among which Example 1 has the strongest antioxidant damage performance, and the effects of Examples 1-4 are better than those of the comparative example. The results show that Example 1 has the best antioxidant properties, indicating that the nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects provided by the present application can play a better role.

[0121] Test Example 4: Anti-UV oxidation efficacy test

[0122] Dilute Examples 1-4 and Comparative Examples 1-3 with DMEM culture medium at a final concentration of 10 μg / mL of active ingredients. HDF cells in the logarithmic growth phase were inoculated in a 96-well plate at 8×103 per well, with a system of 100 μL per well. The outermost circle of the 96-well plate was sealed with 100 μL of PBS. After the cells were completely attached to the wall, they were replaced with complete culture medium containing the sample to be tested. After continuing to culture the cells for 24 hours, the cell activity was detected by the CCK-8 method. The cells cultured with the dilution of the sample to be tested for 24 hours were irradiated with UV for 5 minutes, and the cell activity was detected by the CCK8 method after being cultured in an incubator for 24 hours. The results are shown in Figure 3 .

[0123] Depend on Figure 3The results show that after damage by ultraviolet UVB, the cell viability of the group not medicated was 58.7%, and the cell viability level of the UV group was significantly lower than that of the normal group, indicating that the UV damage HaCaT model was successfully established. Compared with the blank control, the nanoemulsion gels prepared in Examples 1-4 and Comparative Example 1 can significantly improve the viability of UV-damaged human epidermal cells, and the difference is statistically significant (P < 0.01). Among them, Example 1 has the strongest anti-UV damage performance, and the effects of Examples 1-4 are better than those of the comparative example. The results show that Example 1 has the strongest anti-UV damage performance, indicating that the nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects provided in this application can play a better role.

[0124] Test Example 5: Skin patch test

[0125] 50 people participated in the test. Dilute Examples 1-4 and Comparative Examples 1-3 to 1%, take 0.02 mL of each and put it in the small chamber of the spot tester, apply the spot tester with the test substance to the back or forearm of the subject with a low-allergenic tape, and press it lightly with the palm to evenly apply it to the skin for 24 hours. Observe the skin reaction according to the standards in Table 1 30 minutes (after the indentation disappears), 24 hours and 48 hours after removing the test substance spot tester. And record the observation results. The results are shown in Table 3.

[0126] Table 2 Reaction degree, scoring criteria and skin reaction criteria

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[0128]

[0129] Table 3 Human patch test results

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[0131] The patch test was performed to observe whether the samples to be tested (Examples 1-4, Comparative Examples 1-3) induced mild dermatitis locally, thereby determining whether the skin was contact allergic to the tested allergens. The test results in Table 3 show that the nanoemulsion gels prepared in Examples 1-4 and Comparative Examples 1-3 did not cause erythema, severe infiltration, edema, or confluent herpes after being applied to the skin, indicating that the nanoemulsion gels with anti-aging and mitochondrial telomerase activation effects provided by the present application have excellent safety in use.

[0132] Test Example 6: Chicken embryo test

[0133] After removing the eggshell and inner membrane of the air chamber of SPF eggs incubated for 9 to 12 days, use a pipette to draw 0.3 mL of the sample to be tested (Example 1, Example 2, Example 3, Example 4) and slowly drip it into the chorioallantoic membrane (CAM). Observe the reaction changes of the CAM within 5 minutes and take photos to record. The blank control is 0.9% saline, and the positive control is 0.1 mol / L sodium hydroxide. The results are shown in Figure 4 .

[0134] Depend on Figure 4 It can be seen that after Example 1, Example 2, Example 3, and Example 4 acted on the chorioallantoic membrane (CAM) for 5 minutes, there was no bleeding, no coagulation, and no vascular lysis, and the endpoint evaluation method scored 0 points, which determined that Example 1, Example 2, Example 3, and Example 4 were non-irritating.

[0135] The various embodiments in this specification are described in a progressive manner, and the same or similar parts between the various embodiments can be referenced to each other, and each embodiment focuses on the differences from other embodiments.

[0136] The above embodiments are only used to illustrate the technical solutions of the present application, rather than to limit the present application. Although the present application has been described in detail with reference to the aforementioned embodiments, a person of ordinary skill in the art should understand that the technical solutions recorded in the aforementioned embodiments may still be modified, or some or all of the technical features therein may be replaced by 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 present application.

Claims

1. A nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects, characterized in that: It is composed of the following components in percentage by mass: 5-20wt% liposome bilayer membrane flow mosaic regulator, 1-10wt% bilayer film-forming agent, 5-20wt% active ingredient solvent, 0.1-5wt% pterostilbene, 0.1-5wt% cycloastragenol, 0.1-5wt% baicalein, 0.1-5wt% quercetin, 1-10wt% soluble phosphopeptide, 1-5wt% salvianolic acid B, 10-30wt% glycerol, and the balance is ultrapure water; Wherein, the liposome bilayer membrane flow mosaic regulator is at least one of cholesterol, 1,3-propylene glycol, 1,2-pentanediol, 1,2-hexanediol, deacetylated chitosan or yeast β-glucan.

2. The nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects according to claim 1, characterized in that: The bilayer film-forming agent is at least one of lecithin, soybean lecithin, hydrogenated lecithin or egg yolk lecithin.

3. The nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects according to claim 1, characterized in that: The active ingredient solvent is at least one of PEG-8 caprylic triglyceride, PEG-8 capric triglyceride, PEG-40 hydrogenated castor oil, diisopropyl sebacate, polysorbate-20 or polyglyceryl-10 stearate.

4. The nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects according to claim 1, characterized in that: The pterostilbene, cycloastragenol, baicalin, quercetin, soluble phosphopeptide and salvianolic acid B are of natural origin, biosynthetic origin or chemical synthetic origin.

5. A method for preparing the nanoemulsion gel having anti-aging and mitochondrial telomerase activation effects according to any one of claims 1 to 4, characterized in that: The steps include: S1: After the active ingredient solvent and glycerol are mixed to form a mixed solution, pterostilbene, cycloastragenol, baicalein and quercetin are ultrasonically dispersed in the mixed solution to obtain phase A; S2: After mixing the liposome bilayer membrane flow mosaic regulator, bilayer membrane-forming agent, soluble phosphopeptide and ultrapure water, the mixture is stirred under heating conditions in a water bath to obtain phase B; S3: under shearing conditions, adding phase A to phase B for mixing, homogenizing and shearing to obtain liposome colostrum; S4: subjecting the liposome colostrum to ultra-high pressure homogenization and cooling it to room temperature to obtain a liposome emulsion; S5: At room temperature, salvianolic acid B is added to the liposome emulsion, solubilized with ultrasonic aid and incubated overnight to obtain a nanoemulsion gel.

6. The preparation method according to claim 5, characterized in that: In step S2, the water bath heating temperature is 40-50°C, and the stirring speed is 200-500 rpm.

7. The preparation method according to claim 5, characterized in that: In step S3, the shearing speed is 5000-8000 rpm, the shearing temperature is 40-50°C, and the shearing time is 5-10 min.

8. The preparation method according to claim 5, characterized in that: In step S4, the pressure is 500-1000 bar, the temperature is 40-80°C, and the cycle is 6-9 times; in step S5, the ultrasonic power is 80-100w, the dissolution time is 30-40min, and the incubation temperature is 4°C.

9. Use of the nanoemulsion gel with anti-aging and mitochondrial telomerase activation effects according to claim 1 in the preparation of cosmetics.

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