Anti-aging composition and use thereof

Abstract

The application discloses a kind of anti-aging composition, including active substance and thioneine, and the active substance includes vitamin or its derivatives and amino acid or its derivatives.Through active substance and thioneine synergistic action, there is significant anti-aging and antioxidant effect, while all have good effect in promoting collagen generation, comprehensively improve anti-aging effect.The anti-aging composition of the present application can be used for various skin care products, can be used alone as anti-aging product and can also play a role together with other ingredients such as moisturizing, whitening, improve the comprehensive effect of skin care products, with diversified purposes.

Description

Technical Field

[0001] The present application relates to the field of fine chemicals, and in particular to an anti-aging composition and its use. Background Art

[0002] The skin is the largest organ in the human body, providing a protective barrier and therefore crucial for maintaining homeostasis. With age, the body's antioxidant capacity gradually decreases. Ionizing radiation, ultraviolet radiation, foreign chemicals, and medications can also cause the body to produce large quantities of reactive oxygen free radicals. Normally, the body has a system to combat free radicals, protecting it from damage. However, with age, free radicals increase in the body, and the body's ability to combat them decreases. The balance between free radical production and elimination becomes unbalanced, and normal tissues are attacked and damaged by free radicals, impairing their function and causing skin aging. This in turn leads to oxidative stress and damage, causing inflammation, dryness, sagging, wrinkles, and pigmentation, accelerating the aging process.

[0003] Skin aging is a complex and slow process, and its main causes can be divided into two aspects: one is the physiological aging of the skin, in which DNA replication is inhibited and the expression of aging genes is critical; the other is exogenous environmental factors, among which ultraviolet rays have the greatest impact on the skin.

[0004] Therefore, taking in antioxidants that can fight free radicals from the outside world has become a way to protect skin cells from damage. Good anti-oxidation and scavenging of free radicals can play an anti-aging role and avoid premature aging of the skin. Therefore, skin anti-oxidation is increasingly valued by the cosmetics industry. Anti-oxidation once became synonymous with anti-aging, and the application of antioxidants is of great significance in the cosmetics industry. Summary of the Invention

[0005] The present application utilizes the excellent antioxidant function of thioneine to prepare an antioxidant and anti-aging composition. At the same time, in order for thioneine to better play its role, the present application provides a specific ecological environment for thioneine, wherein the active substance is combined with thioneine, which is more conducive to the performance of the antioxidant function of thioneine. At the same time, the active substance is combined with thioneine, which has a synergistic anti-aging effect and also has the effect of stimulating and promoting collagen production.

[0006] The present application provides an anti-aging composition, comprising active substances and ergothioneine, wherein the active substances include vitamins or derivatives thereof and amino acids or derivatives thereof.

[0007] The mass ratio of the active substance to ergothioneine is 1:(0.0005-2).

[0008] The mass ratio of the vitamin or its derivative to the amino acid or its derivative is 1:(0.005-20).

[0009] The present application also provides a method for preparing the above composition, which comprises dissolving the above active substance and ergothioneine in a solvent to prepare the composition.

[0010] The solvent includes an aqueous solvent.

[0011] The present application also provides the use of the above composition in anti-aging.

[0012] The present application also provides use of the above composition in anti-oxidation.

[0013] The present application also provides use of the above composition in skin care products.

[0014] The present application also provides a skin care product comprising the above composition.

[0015] Effects of the Invention

[0016] 1. the anti-aging composition of the application, by active substance and thioneine synergistic action, has significant anti-aging and antioxidant effect, is better than thioneine and is used alone. Simultaneously in promoting collagen generation, all have good effect, comprehensively improve anti-aging effect.

[0017] 2. The anti-aging composition of the present application can be used in various skin care products. It can be used alone as an anti-aging product or can work together with other moisturizing, whitening and other ingredients to enhance the overall efficacy of skin care products and has diversified uses. DETAILED DESCRIPTION

[0018] The following description of exemplary embodiments of the present application includes various details of the embodiments of the present application to facilitate understanding, and should be considered as merely exemplary. Therefore, those skilled in the art will recognize that various changes and modifications may be made to the embodiments described herein without departing from the scope and spirit of the present application. Similarly, for the sake of clarity and conciseness, descriptions of well-known functions and structures are omitted in the following description.

[0019] The present application provides an anti-aging composition comprising an active substance and ergothioneine.

[0020] The present inventor surprisingly found through research that vitamins or their derivatives and amino acids or their derivatives as active substances can make ergothioneine play a stronger anti-aging and antioxidant effect.

[0021] Ergothioneine (ERG), formally known as 2-mercapto L-histidine trimethyl inner salt, has multiple functions such as anti-oxidation, anti-inflammatory and anti-ultraviolet radiation, and is a natural antioxidant. After being rapidly absorbed by the human body or other animals from the diet, ergothioneine is not rapidly metabolized or excreted into the urine, but is present in human tissues and body fluids in the form of thioketones, and is therefore not prone to auto-oxidation and can accumulate to a higher level in certain tissues and body fluids. Its strong in vitro antioxidant properties and natural safety make ergothioneine highly valuable for development and utilization in various fields such as food additives, health care pharmaceuticals, and cosmetics and skin care. Literature shows that factors such as extraction method, purification method, and physicochemical environment all have an impact on the in vitro antioxidant properties and stability of ergothioneine.

[0022] The composition provided by the present application contains active substances, and the in vitro antioxidant properties and stability of ergothioneine are enhanced through the activation of the active substances.

[0023] In some embodiments, the mass ratio of the active substance to ergothioneine is 1: (0.0005-2), for example, 1: 0.0005, 1: 0.0006, 1: 0.0007, 1: 0.0008, 1: 0.0009, 1: 0.001, 1: 0.005, 1: 0.01, 1: 0.015, 1: 0.02, 1: 0.025, 1: 0.03, 1: 0.035, 1: 0.04, 1: 0.045, 1: 0.05, 1: 0.055, 1:0.06, 1:0.065, 1:0.07, 1:0.075, 1:0.08, 1:0.085, 1:0.09, 1:0.095, 1:0.1, 1:0.2, 1:0.3, 1:0.4, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1.0, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2.0.

[0024] In some preferred embodiments, the mass ratio of the active substance to ergothioneine is 1:(0.001-1).

[0025] In some preferred embodiments, the mass ratio of the active substance to ergothioneine is 1:(0.01-0.5).

[0026] In some embodiments, the mass ratio of the vitamin or its derivative to the amino acid or its derivative is 1:(0.005-20), for example, 1:0.005, 1:0.006, 1:0.007, 1:0.008, 1:0.009, 1:0.01, 1:0.015, 1:0.02, 1:0.025, 1:0.03, 1:0.035, 1:0.04, 1:0.045, 1:0.05, 1:0.055, 1:0.06, 1:0.065, 1:0. 07, 1:0.075, 1:0.08, 1:0.085, 1:0.09, 1:0.095, 1:0.1, 1:0.2, 1:0.3, 1:0.4, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20.

[0027] In some preferred embodiments, the mass ratio of the vitamin or its derivative to the amino acid or its derivative is 1:(0.01-10).

[0028] In some preferred embodiments, the mass ratio of the vitamin or its derivative to the amino acid or its derivative is 1:(0.1-5).

[0029] In some embodiments, the amount of the active substance added is 0.1-5wt%, preferably 0.5-2wt%, based on the total percentage of the composition and the solvent, for example, it can be 0.1wt%, 0.2wt%, 0.3wt%, 0.4wt%, 0.5wt%, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, or 5wt%.

[0030] In some embodiments, in the total percentage of the composition and the solvent, the addition amount of the thioneine is 0.0001-5wt%, preferably 0.001-1wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt%, 0.0006wt%, 0.0007wt%, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005 wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08 wt%, 0.09wt%, 0.1wt%, 0.2wt%, 0.3wt%, 0.4wt%, 0.5wt%, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%.

[0031] In some embodiments, the added amount of the vitamin or its derivative is 0.1-5wt%, preferably 0.5-2wt%, based on the total percentage of the composition and the solvent, for example, it can be 0.1wt%, 0.2wt%, 0.3wt%, 0.4wt%, 0.5wt%, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%.

[0032] In some embodiments, the amount of the amino acid or its derivative added is 0.1-5wt%, preferably 0.5-2wt%, based on the total percentage of the composition and the solvent, for example, it can be 0.1wt%, 0.2wt%, 0.3wt%, 0.4wt%, 0.5wt%, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, or 5wt%.

[0033] The vitamins or their derivatives include but are not limited to vitamin A or its derivatives, vitamin B group or its derivatives, vitamin C or its derivatives, vitamin D or its derivatives, vitamin E or its derivatives, vitamin K or its derivatives, etc. In some preferred embodiments, the present application selects vitamin B group and its derivatives, vitamin C or its derivatives. In some preferred embodiments, the present application selects vitamin B group and its derivatives, vitamin C or its derivatives. In some preferred embodiments, the vitamins selected in the present application are vitamin B group and its derivatives, for example, vitamin B1 (thiamine) or its derivative thiamine hydrochloride, vitamin B2 (riboflavin), vitamin B3 (niacin) or its derivative niacinamide, vitamin B4 (adenine), vitamin B5 (pantothenic acid) or its derivative panthenol, vitamin B6 (pyridoxine) or its derivative pyridoxine hydrochloride, vitamin B7 (biotin), vitamin B9 (folic acid), vitamin B12 (cyanocobalamin), etc.

[0034] In the present application, the amino acid refers to a class of compounds containing both amino and carboxyl groups in a molecule, and amino acid derivatives are substances formed by a series of reaction compounds of amino acids, i.e., the predecessor of amino acid derivatives is amino acid. It should be emphasized that in the present application, the amino acid and derivatives thereof refer to any other amino acids and derivatives thereof not comprising thioneine.

[0035] The amino acids or their derivatives include, but are not limited to, histidine or histidine hydrochloride, isoleucine, leucine, lysine or lysine hydrochloride, polylysine, methionine or acetylmethionine, phenylalanine, threonine, tryptophan, valine, alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine or glycine salt, proline or proline hydrochloride, serine or methylserine and tyrosine, etc.

[0036] In some preferred embodiments, the amino acids include one or more selected from proline, methionine, leucine or glycine.

[0037] In some embodiments, the amino acid or its derivative includes one or more selected from methionine, proline or glycine.

[0038] In some embodiments, the amino acid or its derivative is methionine.

[0039] In some embodiments, the amino acid or its derivative is proline.

[0040] In some embodiments, the amino acid or its derivative is glycine or a glycine salt.

[0041] In some embodiments, the amino acid or its derivative is leucine.

[0042] In some embodiments, the amino acids or their derivatives are methionine and proline.

[0043] In some embodiments, the amino acids or their derivatives are methionine and glycine.

[0044] In some embodiments, the amino acids or derivatives thereof of the present application are methionine and leucine.

[0045] In some embodiments, the amino acids or their derivatives are proline and glycine.

[0046] In some embodiments, the amino acid or its derivative is methionine, proline and glycine.

[0047] In some embodiments, the amino acids or derivatives thereof of the present application are methionine, proline and leucine.

[0048] In some embodiments, the amino acids or derivatives thereof of the present application are methionine, leucine and glycine.

[0049] In some embodiments, the amino acids or derivatives thereof of the present application are glycine, proline and leucine.

[0050] In some embodiments, the amino acid or its derivatives of the present application are methionine, glycine, proline and leucine. In some embodiments, the active substance includes adenine and proline.

[0051] In some embodiments, the active substance of the present application includes adenine and glycine.

[0052] In some embodiments, the active substance of the present application includes adenine and methionine.

[0053] In some embodiments, the active substance of the present application includes adenine and leucine.

[0054] In some embodiments, the active substance includes biotin and proline.

[0055] In some embodiments, the active substance of the present application includes biotin and glycine.

[0056] In some embodiments, the active substance of the present application includes biotin and methionine.

[0057] In some embodiments, the active substance of the present application includes biotin and leucine.

[0058] In some embodiments, the active substance of the present application includes vitamin C and proline.

[0059] In some embodiments, the active substance of the present application includes vitamin C and glycine.

[0060] In some embodiments, the active substance of the present application includes vitamin C and methionine.

[0061] In some embodiments, the active substance of the present application includes vitamin C and leucine.

[0062] In some preferred embodiments of the present application, any of the compositions described above is composed of an active substance and ergothioneine.

[0063] In some preferred embodiments of the present application, the active substance consists of vitamins or their derivatives and amino acids or their derivatives.

[0064] The present application provides use of any of the compositions described above in anti-aging.

[0065] The present application also provides use of any of the above compositions in skin care products.

[0066] The applications include, but are not limited to, adding the composition provided by the present application to skin care products, directly combining the composition with skin care products, and indirectly combining the composition with other skin care products.

[0067] The skin care products may be in liquid, semi-solid, or solid form, including but not limited to essences, sunscreens, oils, body toners, bath products, eye area skin care products, facial masks, facial cleansers, makeup removers, foundations, pressed powders, rouges, body makeup, eyebrow pencils, eyeshadows, eyelid makeup, eye hair makeup, eye makeup removers, lip balms, lip glosses, lipsticks, lip liners, etc.

[0068] The present application further provides a skin care product, comprising any one of the compositions described above.

[0069] In some embodiments, the skin care product may further include antioxidants, preservatives and mildew inhibitors, moisturizers, surfactants, pH regulators, fragrances, pigments, functional additives and other components.

[0070] The anti-aging composition of the present application has excellent anti-aging effects by combining active substances with ergothioneine. For example, in promoting collagen synthesis, the anti-aging composition of the present application can significantly improve collagen synthesis, and its collagen production rate can reach up to 161%, which is nearly 50% higher than that of compositions other than the present application. In addition, the synergistic effect between the various substances in the composition of the present application is significant, and the Q value calculated by King's formula can reach up to 1.48, which has a very obvious synergistic enhancement effect.

[0071] And oxidation, a major factor of skin aging, is also what the composition of the present application can effectively resist. For example, diphenyl picrohydrazine free radical (DPPH) is a stable nitrogen-centered free radical. Its amount is an important indicator for judging the degree of oxidation. The composition of the present application has a significant improvement in DPPH clearance rate. The present application provides an environment by providing a suitable active substance for ergothioneine, which can make ergothioneine play a more outstanding free radical scavenging effect. Compared to only containing the same amount of ergothioneine without active substance, the composition of the present application improves DPPH clearance rate by nearly 10%.

[0072] For example, lipid peroxidation is one of the cell pathways involved in oxidative damage, and the compositions of the present application can effectively remove malondialdehyde (MDA), one of the final products of lipid peroxidation, so as to improve the situation of lipid peroxidation, and then alleviate oxidative process and reach anti-aging purpose. The compositions of the present application, in the presence of active substance, thioneine plays the clearing effect of more outstanding MDA, and compared to and without active substance, the compositions MDA clearance rate only containing thioneine can be improved by more than 15%.

[0073] Therefore, the composition of the present application has substantial improvements in promoting collagen synthesis and anti-oxidation, both of which are important factors in delaying aging. Therefore, the composition of the present application has a very good anti-aging effect and can be applied to a variety of skin care products.

[0074] Example

[0075] Example 1

[0076] 0.5 g of adenine, 0.5 g of proline, and 0.001 g of ergothioneine were accurately weighed, dissolved in purified water, and the volume was adjusted to 100 ml to obtain a composition.

[0077] Examples 2-16 and Comparative Examples 1-8

[0078] The compositions of Examples 2-16 and Comparative Examples 1-8 were prepared according to the method of Example 1, wherein the contents of amino acids, vitamins and thioneine are as shown in Table 1.

[0079] Table 1

[0080]

[0081]

[0082] Test Example 1 Effect of Promoting Type I Collagen Production

[0083] Reagents and instruments

[0084] DMEM basal culture medium (Gibco), fetal bovine serum (Gibco), PBS (Gibco), 0.25% trypsin-EDTA containing phenol red (Gibco), penicillin-streptomycin double antibody (Gibco), ultrapure water, human type I collagen Human COL-1 ELISA Test Kit (Sigma-Aldrich).

[0085] Ultrapure water instrument (Millipore, USA), small high-speed centrifuge (Eppendorf, Germany), vortexer (SCILOGEX, USA), carbon dioxide cell culture incubator (Thermo Fisher, USA), biological safety cabinet (Thermo Fisher, USA), ordinary inverted microscope (Nikon, Japan), electric constant temperature water bath (Shanghai Yiheng Technology Co., Ltd., China), multifunctional microplate reader (Tecan, Switzerland), and UV irradiator (Thermo Fisher, USA).

[0086] Testing Process

[0087] (1) Cell culture

[0088] Human skin fibroblasts (HSF) were cultured in DMEM basal medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C in a carbon dioxide incubator. The cells were passaged at a ratio of 1:2 every three days, and cells of the third generation or above were selected for testing.

[0089] (2) Sample dissolution

[0090] The compositions of the examples and comparative examples were diluted 20-fold respectively with DMEM basal culture medium to obtain sample solutions.

[0091] (3) Detection of human type I collagen COL-1 expression

[0092] HSF was digested with trypsin and 1×10 5 Cells were seeded at a density of 100 cells / mL in a 6-well plate, with 3 mL of cell suspension per well. After overnight incubation in a cell culture incubator, cells were slowly washed once with PBS buffer and 3 mL of sample solution was added. An equal volume of DMEM basal medium was added to the negative control group. After 48 hours, cells were harvested and lysed by sonication for 30 seconds. The supernatant was collected by centrifugation and assayed for COL-1 content according to the instructions of the COL-1 ELISA KIT kit.

[0093] (4) Calculation method of type I collagen production rate

[0094] Collagen production rate = (collagen content of sample group - collagen content of negative control group) / collagen content of negative control × 100%

[0095] The specific test results are shown in Table 2 below.

[0096] We verified the additive effect of the combination using King's formula: Q = E(a+b) / (Ea+Eb-Ea×Eb) (where Q < 0.55 is significantly antagonistic, Q = 0.55-0.85 is antagonistic, Q = 0.85-1.15 is additive, and Q > 1.15 is enhancing).

[0097] For example, when the concentration of thioneine is 0.001%, the type I collagen production rate is 99.78% (Comparative Example 1), when the concentration of adenine is 0.5% and the concentration of proline is 0.5%, the type I collagen production rate is 110.87% (Comparative Example 8), the type I collagen production rate used in combination with thioneine (0.001%) and adenine (0.5%)+proline (0.5%) is 121.52% (Example 1), then,

[0098] Ea = 0.001% ergothioneine type I collagen production rate = 99.78%;

[0099] Eb = 0.5% adenine + 0.5% proline type I collagen production rate = 110.87%;

[0100] E(a+b) = 0.001% ergothioneine combined with 0.5% adenine + 0.5% proline. Type I collagen production rate = 121.52%,

[0101] Substituting this into the formula we get Q = 1.21.

[0102] The Q values ​​of the above concentration combinations were obtained using the same formula. The specific test results are shown in Table 2 below.

[0103] Table 2

[0104]

[0105]

[0106] As can be seen from the Q values ​​calculated by King's formula in Table 2, taking Examples 1-5 of the present application as an example, their Q values ​​are all above 1.15, and the highest can reach 1.48, which shows that the composition of the present application has achieved the effect of synergistically enhancing the production of type I collagen.

[0107] The compositions in the present application have a type I collagen production rate of more than 118%, and can reach up to 161%. Compared with Comparative Example 8 containing only active substances, the collagen production rate is 110.87%, and the collagen production rates of Comparative Examples 1-5 containing only ergothioneine are all around 100%. The composition of the present application has a significantly improved type I collagen production rate. Even if it contains one of the active substances such as vitamins or their derivatives, amino acids or their derivatives, such as Comparative Examples 6 and 7, the collagen production rates are only 114.65% and 99.83%, which are not as good as the compositions of the present application.

[0108] Test Example 2 Antioxidant Effect DPPH Test

[0109] Diphenylpicrohydrazine (DPPH) is a stable nitrogen-centered free radical. A methanol or ethanol solution of DPPH is purple, with maximum absorption at a wavelength of 510-530 nm. Its concentration is linearly related to absorbance. In the presence of a free radical scavenger, the free radical scavenger provides one electron to pair with the lone pair of electrons in DPPH, causing it to fade. The degree of fading is quantitatively related to the electrons received, manifested as a lighter color of the solution and a decrease in absorbance. The stronger the free radical scavenger, the lower the absorbance. Accurately measure 5.0 mL of DPPH solution and 5.0 mL of sample solution into stoppered test tubes and mix thoroughly. Use an equal volume of a mixed solution of water and 95% ethanol as a blank control. Leave at room temperature for 30 minutes, and measure the absorbance of the solution at 523 nm.

[0110] Reagents and instruments

[0111] Experimental materials: 2,2-diphenyl-1-picrylhydrazyl (Aladdin Reagent Shanghai Co., Ltd.), anhydrous ethanol (Sinopharm Chemical Reagent Co., Ltd.)

[0112] Instruments: UV-visible spectrophotometer, UV2550, Shimadzu; AL-104 electronic balance, Mettler-Toledo, Switzerland; pH meter, Mettler-Toledo, Switzerland.

[0113] Testing Process

[0114] (1) Solution preparation

[0115] 0.1 mM DPPH solution: Accurately weigh 4.0 mg of DPPH into a 100 mL brown volumetric flask, dissolve it in 95% ethanol and make up to volume.

[0116] Sample solution: Compositions of Examples 1 to 16 and Comparative Examples 1 to 8

[0117] (2) Operation steps

[0118] Precisely measure 5.0 mL of DPPH solution and 5.0 mL of sample solution into stoppered test tubes and mix thoroughly. Use an equal volume of a mixture of water and anhydrous ethanol as a blank control. Incubate at room temperature for 30 minutes and measure the absorbance of each solution at 523 nm. In a separate group, precisely measure 5.0 mL of DPPH solution and 5.0 mL of purified water and mix them. Repeat the same procedure.

[0119] (3) Calculation method

[0120]

[0121] Specific test results are shown in Table 3 below. As shown in the data in the table, the compositions of the present application are such as Examples 1-5, and Comparative Examples 1-5 containing an equal amount of ergothioneine, the compositions of the present application contain active compositions, do not contain active substances relative to Comparative Examples 1-5, and DPPH clearance is significantly improved, such as the DPPH clearance of Example 4 of the present application is 28.43%, and the DPPH clearance of Comparative Example 4 is 18.89%, which has increased by nearly 10%.

[0122] Table 3

[0123]

[0124]

[0125] Test Example 3: Lipid Peroxidation Test

[0126] Lipid peroxidation is one of the cellular pathways involved in oxidative damage and is closely related to DNA damage, as many of its reaction end products can interact with DNA and cause oxidative DNA damage. It has also recently been found to pose a certain risk of chemical carcinogenesis. Malondialdehyde (MDA) is a biomarker of oxidative damage caused by exposure to environmental foreign substances. It is one of the end products of lipid peroxidation and can be detected both in vivo and in vitro. MDA assessment is an important marker of oxidant-induced cytotoxicity. Therefore, MDA measurement can be used to evaluate the anti-free radical efficacy of different chemicals.

[0127] Reagents and instruments

[0128] Experimental materials: human skin fibroblast HSF, DMEM basal medium (Gibco), fetal bovine serum (Gibco), PBS (Gibco), 0.25% trypsin-EDTA containing phenol red (Gibco), penicillin-streptomycin double antibody (Gibco), cell lysate (Biyuntian), MDA detection kit (Biyuntian).

[0129] Instruments and equipment: ultrapure water meter (Millipore, USA), small high-speed centrifuge (Eppendorf, Germany), vortexer (SCILOGEX, USA), carbon dioxide cell culture incubator (Thermo Fisher, USA), biological safety cabinet (Thermo Fisher, USA), ordinary inverted microscope (Nikon, Japan), electric constant temperature water bath (Shanghai Yiheng Technology Co., Ltd., China), multifunctional microplate reader (Tecan, Switzerland).

[0130] Testing Process

[0131] (1) Solution preparation

[0132] Sample solution: the compositions of Examples 1 to 16 and Comparative Examples 1 to 8,

[0133] Other solutions: PBS buffer, 100 μmol / L hydrogen peroxide, and DMEM basal culture medium.

[0134] (2) Operation steps

[0135] HSF was digested with trypsin and 1×10 5 Cells were seeded at a density of 100 μmol / mL in a 12-well plate, with 1 mL of cell suspension per well. After overnight incubation in a cell culture incubator, cells were slowly washed once with PBS buffer. The model group was treated with 100 μmol / L hydrogen peroxide, while the experimental group was treated with 1 mL of sample solution followed by hydrogen peroxide to a final concentration of 100 μmol / L. The negative control group was treated with an equal volume of DMEM basal medium for 16 hours. Each treatment was replicated three times. After incubation, cells were lysed with cell lysis buffer, and the MDA content in the cells was determined using an MDA detection kit.

[0136] (3) Calculation method

[0137] MDA clearance rate = (MDA concentration in the hydrogen peroxide group - MDA concentration in the sample group) / MDA concentration in the hydrogen peroxide group × 100%.

[0138] Specific test results are shown in Table 4 below. As can be seen from the table, the compositions of the present application, such as Examples 1-5, and Comparative Examples 1-5, containing an equal amount of ergothioneine, contain active compositions in the compositions of the present application, do not contain active substances relative to Comparative Examples 1-5, and have a significant improvement in MDA clearance, such as the MDA clearance of Example 3 of the present application is 47.46%, and the MDA clearance of Comparative Example 3 is 30.87%, which has increased by more than 15%.

[0139] Table 4

[0140]

[0141]

[0142] Although the embodiments of the present application are described above, the present application is not limited to the above-mentioned specific embodiments and application fields. The above-mentioned specific embodiments are merely illustrative and instructive, and not restrictive. A person of ordinary skill in the art, under the guidance of this specification and without departing from the scope of protection of the claims of this application, can also make many forms, all of which fall within the scope of protection of this application.

Claims

1. An anti-aging composition, characterized in that Includes the following components: (1) Vitamin B4; (2) Proline; (3) Ergothioneine; The mass ratio of the sum of the component (1) and the component (2) to ergothioneine is 1: (0.001-1), The mass ratio of the component (1) to the component (2) is 1:(0.01-20).

2. A method for preparing the composition according to claim 1, characterized in that: The composition comprises dissolving component (1), component (2) and ergothioneine in a solvent to prepare the composition. The preparation method according to claim 2 , wherein the solvent comprises an aqueous solvent.

4. Use of the composition according to claim 1 in preparing anti-aging skin care products.

5. Use of the composition according to claim 1 in preparing an antioxidant skin care product.

6. Use of the composition according to claim 1 in preparing a skin care product for promoting collagen production.

7. A skin care product comprising the composition according to claim 1.

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