Cosmetic composition with multi-cell energy improving effect and application thereof

Abstract

The invention discloses a cosmetic composition with a multi-cell energy improving effect and application of the cosmetic composition. The cosmetic composition is obtained by compounding the ribose, the artemia extract and the fructose trisodium diphosphate, the components have a remarkable synergistic effect, and oxidation resistance can be achieved after compounding; mitochondria is protected; promoting skin barrier generation or enhancing skin barrier defense; preserving moisture; wrinkle resistance; therefore, the effect of integrating multiple effects in a real sense is realized, and the use requirements of various types of consumer groups are met.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and particularly relates to a cosmetic composition with multiple cell energy enhancing effects and its application. Background Art

[0002] Traditional cosmetic compositions have long been limited by the problem of single efficacy. The root cause lies in the simple superposition of active ingredients and the lack of synergistic effects. Most existing cosmetics achieve basic efficacy coverage by mechanically mixing single-functional ingredients. However, the physicochemical property differences of different components are likely to cause activity loss or mutual antagonism, resulting in the so-called "multiple effects in one" actually only being able to play a partial role. For example, heat-sensitive ingredients such as vitamin C and retinol are difficult to balance stability and transdermal efficiency in a conventional formulation system, resulting in insufficient efficacy persistence. At the same time, there is also a problem in the prior art that it is difficult to break through the compatibility barrier between water-soluble (such as hyaluronic acid) and fat-soluble (such as squalane) active substances, resulting in the inability to simultaneously achieve multiple functions such as antioxidant, repair, and anti-aging. Although the industry has tried to develop multifunctional ingredients through synthetic biotechnology (such as recombinant collagen) or the compounding of natural active ingredients (such as Ganoderma lucidum extract), it still faces bottlenecks such as imperfect efficacy evaluation systems and a sharp increase in industrialization costs. Of course, there have also been attempts to use multiple emulsion carriers to achieve precise partition encapsulation and staged release of multiple functions such as moisturizing, anti-wrinkle, and whitening by constructing an oil / water / oil (O / W / O) layered structure. However, its production cost is 3 - 5 times higher than that of traditional formulations, which greatly limits its market popularization process.

[0003] Therefore, there is an urgent need to develop a simple cosmetic composition with multiple effects, which can be used as an efficacy combination in various cosmetics to achieve the true "multiple effects in one" effect. Summary of the Invention

[0004] The present invention aims to at least solve the above technical problems existing in the prior art. For this reason, the object of the present invention is to provide a cosmetic composition with multiple effects and its application. The composition in the present invention realizes the co-play of multiple effects by compounding ribose, artemia extract, and sodium fructose diphosphate, and there is a good synergistic effect between the components, which can significantly promote the generation of the skin barrier or enhance the skin barrier defense; moisturize; anti-wrinkle; tighten the skin; antioxidant; protect mitochondria or improve mitochondrial function, etc.

[0005] In the first aspect of the present invention, a composition is provided, and the composition includes: ribose, artemia extract, and sodium fructose diphosphate.

[0006] In some embodiments of the present invention, in the composition, the mass ratio of ribose, artemia extract and trisodium fructose diphosphate is 0.01-0.5:0.01-0.2:0.001-0.01.

[0007] In some embodiments of the present invention, in the composition, the mass ratio of ribose, artemia extract and trisodium fructose diphosphate is 0.03-0.3:0.01-0.15:0.001-0.01.

[0008] In some embodiments of the present invention, in the composition, the mass ratio of ribose, artemia extract and trisodium fructose diphosphate is 0.03:0.018:0.0012.

[0009] In some embodiments of the present invention, in the composition, the mass ratio of ribose, artemia extract and trisodium fructose diphosphate is 0.25:0.15:0.01.

[0010] In some embodiments of the present invention, in the composition, the mass ratio of ribose, artemia extract and trisodium fructose diphosphate is 0.05:0.03:0.002.

[0011] In some embodiments of the present invention, the composition further includes excipients acceptable in cosmetics.

[0012] In some embodiments of the present invention, the excipients acceptable in cosmetics include but are not limited to: solvents, emulsifiers, stabilizers, thickeners, preservatives, fragrances, pigments, fillers and humectants.

[0013] In some embodiments of the present invention, the preservatives include but are not limited to phenoxyethanol and parabens.

[0014] In some embodiments of the present invention, the humectants include but are not limited to glycerol and hyaluronic acid;

[0015] In some embodiments of the present invention, the excipients acceptable in cosmetics further include antioxidants such as vitamin E and butylated hydroxytoluene; chelating agents such as sodium ethylenediaminetetraacetate; surfactants such as sodium lauryl sulfate and cocamidopropyl betaine; pH regulators such as citric acid and sodium lactate.

[0016] In some embodiments of the present invention, the composition further includes other active ingredients for cosmetics.

[0017] In some embodiments of the present invention, the active ingredients for cosmetics refer to substances having at least one of the following functions (1)-(6):

[0018] (1) Promote the generation of skin barrier or enhance the skin barrier defense;

[0019] (2) Moisturize;

[0020] (3) Anti-wrinkle;

[0021] (4) Tighten the skin;

[0022] (5) Antioxidant;

[0023] (6) Protect mitochondria or improve mitochondrial function.

[0024] The second aspect of the present invention provides a preparation method of the composition described in the above aspect, including the following steps:

[0025] Mix ribose, artemia extract and sodium fructose diphosphate according to the mass ratio described in the above aspect to obtain the composition.

[0026] The third aspect of the present invention provides a cosmetic, which contains the composition described in the above aspect.

[0027] In some embodiments of the present invention, in the cosmetic, calculated by the total mass of the cosmetic, the proportion of the composition is 0.6 - 5%.

[0028] In some embodiments of the present invention, the cosmetic may further include excipients acceptable in the cosmetic.

[0029] In some embodiments of the present invention, the excipients acceptable in the cosmetic include but are not limited to: solvents, emulsifiers, stabilizers, thickeners, preservatives, fragrances, pigments, fillers and humectants.

[0030] In some embodiments of the present invention, the preservatives include but are not limited to phenoxyethanol and parabens.

[0031] In some embodiments of the present invention, the humectants include but are not limited to glycerol and hyaluronic acid;

[0032] In some embodiments of the present invention, the excipients acceptable in the cosmetic further include antioxidants such as vitamin E and butylated hydroxytoluene; chelating agents such as sodium ethylenediaminetetraacetate; surfactants such as sodium lauryl sulfate and cocamidopropyl betaine; pH regulators such as citric acid and sodium lactate.

[0033] The fourth aspect of the present invention provides the application of the composition described in the above aspect in the preparation of cosmetics or drugs.

[0034] In some embodiments of the present invention, the cosmetic or drug has at least one of the following functions (1)-(6):

[0035] (1) Promote the generation of skin barrier or enhance the skin barrier defense;

[0036] (2) Moisturize;

[0037] (3) Anti-wrinkle;

[0038] (4) Tighten the skin;

[0039] (5) Antioxidant;

[0040] (6) Protect mitochondria or improve mitochondrial function.

[0041] In some embodiments of the present invention, in the cosmetics or drugs, calculated by total mass, the proportion of the composition is 0.6-5%.

[0042] In some embodiments of the present invention, the cosmetics or drugs are topical preparations.

[0043] In some embodiments of the present invention, the dosage forms of the cosmetics include: emulsions, aqueous solutions, oils, gels and powders.

[0044] In some embodiments of the present invention, the dosage forms of the drugs include: ointments, aerosols and patches.

[0045] The beneficial effects of the present invention are:

[0046] The present invention provides a cosmetic composition with multiple effects, and there is a significant synergistic effect among its components, which can simultaneously produce multiple effects such as antioxidant; protecting mitochondria; promoting the generation of skin barrier or enhancing skin barrier defense; moisturizing; anti-wrinkle; tightening the skin, etc., thereby realizing the true "multi-effect in one" and meeting the usage needs of various types of consumer groups. Description of the Drawings

[0047] Figure 1 ATP content after treatment of different experimental groups.

[0048] Figure 2 Ratio of red / green average fluorescence intensity of mitochondrial membrane potential after treatment of different experimental groups.

[0049] Figure 3 Relative expression level of FLG gene after treatment of different experimental groups.

[0050] Figure 4 For NAD + / NADH ratio.

[0051] Figure 5 HA content after treatment of different experimental groups.

[0052] Figure 6 The relative expression levels of the AQP3 gene after treatment with different experimental groups.

[0053] Figure 7 The improvement of autophagy effect after treatment with different experimental groups.

[0054] Figure 8 The collagen content after treatment with different experimental groups. Detailed implementation manners

[0055] The content of the present invention will be further described in detail below through specific examples. The raw materials, reagents or devices used in the examples and comparative examples can be obtained from conventional commercial channels or by existing technical methods without special instructions. Unless otherwise specified, the test or measurement methods are conventional methods in the art.

[0056] In the following examples, ribose, Artemia extract, and sodium fructose diphosphate are all commercially available products. Of course, those skilled in the art can also prepare the above components based on the preparation methods disclosed in the prior art, which are also included in the protection scope of the present invention.

[0057] Example 1

[0058] In this example, a cosmetic composition with multiple effects is provided, and its composition is as follows:

[0059] By mass, 0.03 parts of ribose, 0.018 parts of Artemia extract, and 0.0012 parts of sodium fructose diphosphate.

[0060] The preparation method is as follows:

[0061] According to the above mass ratio, mix sugar, Artemia extract, and sodium fructose diphosphate evenly to obtain a cosmetic composition with multiple effects.

[0062] Example 2

[0063] In this example, a cosmetic composition with multiple effects is provided, and its composition is as follows:

[0064] By mass, 0.25 parts of ribose, 0.15 parts of Artemia extract, and 0.01 parts of sodium fructose diphosphate.

[0065] The preparation method is as follows:

[0066] According to the above mass ratio, mix sugar, Artemia extract, and sodium fructose diphosphate evenly to obtain a cosmetic composition with multiple effects.

[0067] Example 3

[0068] In this example, a cosmetic composition with multiple effects is provided, and its composition components are as follows:

[0069] By mass, 0.05 parts of ribose, 0.03 parts of Artemia extract, and 0.002 parts of trisodium fructose diphosphate.

[0070] The preparation method is as follows:

[0071] According to the above mass ratio, mix sugar, Artemia extract, and trisodium fructose diphosphate evenly to obtain a cosmetic composition with multiple effects.

[0072] Test Example 1

[0073] In this test example, the energy enhancement effects of Example 1 and single-component substances with corresponding contents were tested. The specific experimental steps are as follows:

[0074] Resuscitate human immortalized epidermal cells (HaCaT) and perform routine culture. When the cell plating rate reaches about 60%, inoculate the cells into a 6-well plate and incubate overnight at 37°C and 5% CO2.

[0075] When the cell plating rate of the 6-well plate reaches about 60%, administer drugs in groups. Conduct experimental grouping: Divide the plated cells into a normal control group (NC) and a sample group. Among them, the normal control group (NC) is not treated with anything and continues to be cultured with fresh medium. The sample group is respectively added with a medium containing the multi-component composite cosmetic composition in Example 1, ribose with the same content, Artemia extract with the same content, or trisodium fructose diphosphate with the same content, and continue to incubate at 37°C and 5% CO2 for 24 h.

[0076] Among them, the addition amount of each substance is: calculated based on the mass of the medium, the proportion of the corresponding substance is:

[0077] Rib+AE+FDP group (i.e., Example 1): 0.03% ribose, 0.018% Artemia extract, and 0.0012% trisodium fructose diphosphate;

[0078] Rib group: 0.03% ribose; AE group: 0.018% Artemia extract; FDP group: 0.0012% trisodium fructose diphosphate.

[0079] After incubation, the culture medium was replaced. The normal control group (NC) was not treated with anything and was continuously cultured with fresh medium for 24 h. In the sample groups, media containing the multi-component composite cosmetic composition in Example 1, ribose with equal content, Artemia extract with equal content, or sodium fructose diphosphate with equal content were added respectively, and incubation was continued for 24 h under the conditions of 37 °C and 5% CO2.

[0080] After incubation, a commercially available kit was used to detect the ATP content.

[0081] The results are shown in Table 1 and Figure 1 as follows.

[0082] Table 1 Results of energy enhancement in each group

[0083]

[0084]

[0085] Among them, compared with NC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; three biological replicates (N = 3).

[0086] ATP (adenosine triphosphate) is a general molecule that directly provides energy within cells and plays a core role in energy metabolism and regulation of physiological functions. When the energy demand of the body increases, ATP releases high-energy phosphate bonds through hydrolysis reactions to provide immediate energy support for the body. Research shows that the synthesis rate and reserve level of ATP are directly related to cell viability and tissue anti-fatigue ability. By detecting the ATP content, the energy metabolism efficiency of an individual can be evaluated, providing a basis for energy enhancement strategies.

[0087] It can be seen from the above results that, compared with the control group (NC), the composition (Rib + AE + FDP) increased the ATP content more significantly than the single components of Rib, AE, and FDP, indicating that the combination of Rib + AE + FDP produced a synergistic effect that was not possessed by the single use of Rib, AE, and FDP.

[0088] Related tests were also carried out on other examples, and it was found that their effects were basically similar to those of the composition in Example 1.

[0089] Test Example 2

[0090] In this test example, the effects of improving mitochondrial viability of Example 2 and the corresponding single-component substances were tested. The specific experimental steps were as follows:

[0091] After resuscitating human dermal fibroblasts, they were cultured routinely. When the cell confluence reached about 60%, the cells were seeded into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0092] When the cell confluence in the 6-well plates reached about 60%, grouping and drug administration were carried out. Experimental grouping: The seeded cells were divided into a normal control group (NC), a model control group (MC), and a sample group. Among them, the normal control group (NC) and the model control group (MC) were not treated with anything and were continued to be cultured with fresh medium. The sample group was respectively added with the medium containing the multi-component composite cosmetic composition in Example 2, equal content of ribose, equal content of Artemia extract, or equal content of sodium fructose diphosphate, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0093] Among them, the addition amount of each substance was: calculated by the mass of the medium, the proportion of the corresponding substance was:

[0094] Rib+AE+FDP group (i.e., Example 2): 0.25% ribose, 0.15% Artemia extract, and 0.01% sodium fructose diphosphate;

[0095] Rib group: 0.25% ribose; AE group: 0.15% Artemia extract; FDP group: 0.01% sodium fructose diphosphate.

[0096] After incubation, the medium was discarded, and the cells were gently rinsed 1-2 times with D-Hanks balanced salt solution (D-HBSS). Then the model control group (MC) and the sample group were exposed to ultraviolet light for 50 s (UVA, 9 mJ / cm 2 ).

[0097] After modeling, the normal control group (NC) and the model control group (MC) were not treated with anything and were continued to be cultured with fresh medium for 24 h. The sample group was respectively added with the above-mentioned medium containing the multi-component composite cosmetic composition in Example 2, equal content of ribose, equal content of Artemia extract, or equal content of sodium fructose diphosphate, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0098] After incubation, JC-1 staining was carried out, and observations and photographs were taken under a fluorescence microscope.

[0099] Image J was used to analyze the fluorescence intensity (S) of each group, and the ratio of the red / green average fluorescence intensity of the mitochondrial membrane potential was calculated (the calculation formula is shown below).

[0100]

[0101] In the formula:

[0102] P represents the average fluorescence intensity ratio of red to green of mitochondrial membrane potential.

[0103] The results are shown in Table 2 and Figure 2 as follows.

[0104] Results of mitochondrial viability improvement in each group in Table 2

[0105] Sample Group Test Result SD P Value Improvement Rate (vs NC) Normal Control (NC) 1.24 0.028 <0.0001**** 86％ Model Control (MC) 0.67 0.053 / / Rib 0.74 0.022 0.1159 10％ AE 0.79 0.017 0.0196* 18％ FDP 0.83 0.018 0.0073** 24％ Rib + AE + FDP 1.11 0.077 0.0013** 65％

[0106] Among them, compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; three biological replicates (N = 3).

[0107] Mitochondrial membrane potential (MMP) is a core indicator of mitochondrial energy conversion efficiency. A decrease in MMP will weaken ATP synthesis, trigger ROS accumulation and mitochondrial dysfunction; while an increase in MMP can enhance the activity of the electron transport chain, promote efficient energy production, reduce oxidative damage and restore mitochondrial dynamic balance, thus directly enhancing mitochondrial viability. Its level can be detected by fluorescence probes, providing a basis for evaluating mitochondrial health and intervening in energy metabolism problems.

[0108] It can be seen from the above results that compared with the control group (MC), the effect of the composition (Rib + AE + FDP) in enhancing mitochondrial membrane potential is more significant than that of the single components of Rib, AE, and FDP, indicating that the combination of Rib + AE + FDP produces a synergistic effect that is not possessed by Rib, AE, and FDP when used alone.

[0109] Related tests were also carried out on other embodiments, and it was found that their effects were basically similar to those of the composition in Example 2.

[0110] Test Example 3

[0111] In this test example, the barrier repair effects of Example 3 and the single-component substances with corresponding contents were tested. The specific experimental steps are as follows:

[0112] After resuscitating human immortalized epidermal cells (HaCaT), they were cultured routinely. When the cell plating rate reached about 60%, the cells were inoculated into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0113] When the cell seeding rate in the 6-well plate reaches about 60%, group drug administration is carried out. Experimental grouping: The seeded cells are divided into a normal control group (NC) and a sample group. Among them, the normal control group (NC) is not treated with anything and is continuously cultured with fresh medium. The sample group is respectively added with a medium containing the multi-component composite cosmetic composition in Example 3, equal content of ribose, equal content of Artemia extract or equal content of sodium fructose diphosphate, and is continuously incubated at 37 °C and 5% CO2 for 24 h.

[0114] Among them, the addition amount of each substance is: based on the mass of the medium, the proportion of the corresponding substance is:

[0115] Rib+AE+FDP group (i.e., Example 3): 0.05% ribose, 0.03% Artemia extract and 0.002% sodium fructose diphosphate;

[0116] Rib group: 0.05% ribose; AE group: 0.03% Artemia extract; FDP group: 0.002% sodium fructose diphosphate.

[0117] After the incubation is completed, the medium is changed. The normal control group (NC) is not treated with anything and is continuously cultured with fresh medium for 24 h. The sample group is respectively added with the above-mentioned medium containing the multi-component composite cosmetic composition in Example 3, equal content of ribose, equal content of Artemia extract or equal content of sodium fructose diphosphate, and is continuously incubated at 37 °C and 5% CO2 for 24 h.

[0118] After the incubation is completed, a commercially available kit is used to extract total RNA, reverse transcribe, and detect the relative expression levels of β-actin (internal reference) and FLG gene based on q-PCR.

[0119] The results are shown in Table 3 and Figure 3 as follows.

[0120] Table 3 Barrier repair results of each group

[0121] Sample Group Test Result SD P Value Enhancement Rate (vs NC) Normal Control (NC) 1.00 0.064 / / Rib 1.34 0.132 0.0169* 33％ AE 1.26 0.229 0.1352 26％ FDP 1.20 0.162 0.1221 20％ Rib + AE + FDP 1.69 0.246 0.0093** 69％

[0122] Among them, compared with NC, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; three biological replicates (N = 3).

[0123] FLG (filaggrin) is a core protein of the skin barrier function, and its expression level directly determines the barrier integrity. The deletion or degradation of FLG will lead to the loosening of the stratum corneum structure, the reduction of natural moisturizing factor (NMF), and the aggravation of water loss, resulting in inflammation and sensitivity; while the enhancement of FLG synthesis can promote the densification of the stratum corneum, improve the moisturizing ability and block external stimuli, thus accelerating the barrier repair. The status of FLG can be evaluated by immunohistochemistry or gene detection, providing a basis for skin care targeting barrier repair.

[0124] It can be seen from the above results that compared with the control group (NC), the relative expression level of the FLG gene was more significantly increased by the composition (Rib+AE+FDP) than by the single components of Rib, AE, and FDP, indicating that the combination of Rib+AE+FDP produced a synergistic effect that was not possessed by the single use of Rib, AE, and FDP.

[0125] Related tests were also carried out on other embodiments, and it was found that their effects were basically similar to those of the composition in Example 3.

[0126] Test Example 4

[0127] In this test example, the effects of Example 1 and the single-component substances with corresponding contents on improving mitochondrial dysfunction were tested. The specific experimental steps were as follows:

[0128] After resuscitating human dermal fibroblasts, they were cultured routinely. When the cell plating rate reached about 60%, the cells were inoculated into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0129] When the cell plating rate of the 6-well plates reached about 60%, grouping and drug administration were carried out. The experimental groups were divided as follows: the plated cells were divided into a normal control group (NC), a model control group (MC), and a sample group. Among them, the normal control group (NC) and the model control group (MC) were not treated with anything and were continued to be cultured with fresh medium. The sample group was respectively added with a medium containing the multi-component composite cosmetic composition in Example 1, ribose with the same content, Artemia extract with the same content, or trisodium fructose diphosphate with the same content, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0130] Among them, the addition amount of each substance was: calculated by the mass of the medium, the proportion of the corresponding substance was:

[0131] Rib+AE+FDP group (i.e., Example 1): 0.03% ribose, 0.018% Artemia extract, and 0.0012% trisodium fructose diphosphate;

[0132] Rib group: 0.03% ribose; AE group: 0.018% Artemia extract; FDP group: 0.0012% sodium fructose diphosphate.

[0133] After incubation, discard the medium and gently wash the cells 1 - 2 times with D-Hanks balanced salt solution (D-HBSS). Then expose the model control group (MC) and the sample groups to ultraviolet light for 50 s (UVA, 9 mJ / cm 2 ).

[0134] After model establishment, the normal control group (NC) and the model control group (MC) were not treated with anything and continued to be cultured with fresh medium for 24 h. The sample groups were respectively added with the above-mentioned media containing the multi-component composite cosmetic composition in Example 1, equal contents of ribose, equal contents of Artemia extract or equal contents of sodium fructose diphosphate, and continued to be incubated for 24 h under the conditions of 37 °C and 5% CO2.

[0135] After incubation, a commercially available kit was used to detect NAD + and NADH, and the NAD + / NADH ratio was calculated.

[0136] The results are shown in Table 4 and Figure 4 as follows.

[0137] Table 4 Improvement effects of mitochondrial dysfunction in each group

[0138] Sample Group Test Result SD P Value Enhancement Rate (vs NC) Normal Control (NC) 11.72 0.875 0.0012** 64％ Model Control (MC) 7.14 0.387 / / Rib 7.95 0.231 0.0435* 11％ AE 7.77 0.209 0.0784 9％ FDP 8.07 0.141 0.0188* 13％ Rib + AE + FDP 9.16 0.393 0.0031** 28％

[0139] Among them, compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; three biological replicates (N = 3).

[0140] The NAD + / NADH ratio affects the ability of cells to produce energy. In the cytoplasm, the conversion of glucose to pyruvate through glycolysis requires NAD + . In the mitochondria, the TCA cycle reduces NAD + molecules to produce multiple NADH molecules, and these NADH are oxidized by Complex I of the electron transport chain (ETC) to generate ATP. A decrease in NAD + levels affects mitochondrial function, overall cell health, and the development of age-related diseases. This test example evaluates whether the sample has the effect of improving mitochondrial dysfunction by detecting the NAD + / NADH ratio level.

[0141] As can be seen from the above results, compared with the control group (MC), the composition (Rib + AE + FDP) increased the content of NAD+ / NADH more significantly than the single components of Rib, AE, and FDP, indicating that the combination of Rib + AE + FDP produced a synergistic effect that was not possessed by the single use of Rib, AE, and FDP.

[0142] Related tests were also carried out on other embodiments, and it was found that their effects were basically similar to those of the composition in Example 1.

[0143] Test Example 5

[0144] In this test example, the moisturizing effects of Example 2 and the single-component substances with corresponding contents were tested. The specific experimental steps were as follows:

[0145] After resuscitating human immortalized keratinocytes (HaCaT), they were cultured routinely. When the cell plating rate reached about 60%, the cells were seeded in a 6-well plate and incubated overnight at 37°C and 5% CO2.

[0146] When the cell plating rate in the 6-well plate reached about 60%, grouping and drug administration were carried out. The experimental groups were divided as follows: the plated cells were divided into a normal control group (NC) and a sample group. Among them, the normal control group (NC) was not treated with anything and continued to be cultured with fresh medium. The sample group was respectively added with a medium containing the multi-component composite cosmetic composition in Example 2, ribose with the same content, Artemia extract with the same content, or trisodium fructose diphosphate with the same content, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0147] Among them, the addition amounts of each substance were as follows: based on the mass of the medium, the proportion of the corresponding substance was:

[0148] Rib + AE + FDP group (i.e., Example 2): 0.25% ribose, 0.15% Artemia extract, and 0.01% trisodium fructose diphosphate;

[0149] Rib group: 0.25% ribose; AE group: 0.15% Artemia extract; FDP group: 0.01% trisodium fructose diphosphate.

[0150] After the incubation ended, the normal control group (NC) was not treated with anything and continued to be cultured with fresh medium for 24 h. The sample group was respectively added with the above-mentioned medium containing the multi-component composite cosmetic composition in Example 2, ribose with the same content, Artemia extract with the same content, or trisodium fructose diphosphate with the same content, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0151] After the incubation, 200 μL of cell culture supernatant was collected from each well, and the content of hyaluronic acid (HA) was detected using a commercially available ELISA kit. The up-regulation rate was calculated according to the following formula.

[0152]

[0153] In the formula:

[0154] H represents the content of hyaluronic acid (HA).

[0155] Meanwhile, total RNA was extracted from each experimental group, cDNA was synthesized, and the gene expressions of β-actin and the target gene were detected by q-PCR. β-actin was used as the internal reference for gene expression, and the relative RNA expression level of the target gene was calculated.

[0156]

[0157] In the formula:

[0158] Q represents the relative expression level of the AQP3 gene.

[0159] The results are shown in Table 5, Table 6 and Figure 5 、 Figure 6 as follows.

[0160] Table 5 HA content of each group

[0161]

[0162]

[0163] Among them, compared with NC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; three biological replicates (N = 3).

[0164] Table 6 Relative expression level of AQP3 gene

[0165] Sample Group Test Result SD P Value Upregulation Rate (vs NC) Normal Control (NC) 1.00 0.059 / / Rib 1.21 0.079 0.0199 21％ AE 1.13 0.062 0.0617 13％ FDP 1.25 0.087 0.0141 25％ Rib + AE + FDP 1.40 0.061 0.0012 40％

[0166] Among them, compared with NC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; three biological replicates (N = 3).

[0167] The effects of the skin moisturizing system on moisturization from the inside out can be summarized in sequence as: water capture, water locking, water activation, and water storage. Hyaluronic acid (HA) in the dermis can absorb a large amount of water, serving as the water reservoir of the skin, providing a reserve for the moisture of the entire skin, and at the same time filling the dermis to make the structure of the dermis more complete. Aquaporin (AQP) exists widely in various tissue parts of organisms and affects the process of water metabolism in organisms. AQP is highly expressed in the plasma membranes of keratinocytes in the epidermal basal and spinous layers and acts as a water and glycerol transport protein in these plasma membranes, promoting skin hydration and being a key factor in maintaining skin hydration. Therefore, in this test example, the moisturizing efficacy of the sample was evaluated by detecting the content of hyaluronic acid (HA) and the relative expression level of the AQP3 gene.

[0168] As can be seen from the above results, compared with the control group (NC), the moisturizing effect improved by the composition (Rib + AE + FDP) was more significant than that of the single components of Rib, AE, and FDP, indicating that a synergistic effect was produced after the combination of Rib + AE + FDP, which was not possessed by the single use of Rib, AE, and FDP.

[0169] Related tests were also carried out on other embodiments, and it was found that their effects were basically similar to those of the composition in Example 2.

[0170] Test Example 6

[0171] In this test example, the autophagy-promoting effects of Example 3 and the single-component substances with corresponding contents were tested. The specific experimental steps were as follows:

[0172] After resuscitating the fibroblasts, they were cultured routinely. When the cell seeding rate reached about 60%, the cells were inoculated into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0173] When the cell seeding rate in the 6-well plates reached about 60%, grouping and drug administration were carried out. The experimental grouping was as follows: the seeded cells were divided into a normal control group (NC) and a sample group. Among them, the normal control group (NC) was not treated with anything and continued to be cultured with fresh medium. The sample group was respectively added with a medium containing the multi-component composite cosmetic composition in Example 3, ribose with the same content, Artemia extract with the same content, or trisodium fructose diphosphate with the same content, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0174] Among them, the addition amounts of each substance were as follows: based on the mass of the medium, the proportions of the corresponding substances were:

[0175] Rib + AE + FDP group (i.e., Example 3): 0.05% ribose, 0.03% Artemia extract, and 0.002% trisodium fructose diphosphate;

[0176] Rib group: 0.05% ribose; AE group: 0.03% Artemia extract; FDP group: 0.002% sodium fructose diphosphate.

[0177] After the incubation was completed, the medium was changed. The normal control group (NC) was not treated with anything and was continuously cultured with fresh medium for 24 h. The sample groups were respectively added with the above-mentioned medium containing the multi-component composite cosmetic composition in Example 3, equal contents of ribose, equal contents of Artemia extract or equal contents of sodium fructose diphosphate, and were continuously incubated for 24 h under the conditions of 37 °C and 5% CO2.

[0178] After the incubation was completed, RNA was extracted using a commercially available kit. After reverse transcription to cDNA, fluorescence quantitative PCR detection was performed.

[0179] The results are shown in Table 7 and Figure 7 as follows.

[0180] Table 7 Autophagy promotion effects of each group

[0181] Sample Group Test Result SD P Value Enhancement Rate (vs NC) Normal Control (NC) 1.00 0.010 / / Rib 1.16 0.036 0.0019** 16％ AE 1.21 0.038 0.0007*** 21％ FDP 1.29 0.032 0.0001*** 29％ Rib + AE + FDP 1.60 0.052 0.0000**** 59％

[0182] Among them, compared with NC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; three biological replicates (N = 3).

[0183] LC3B (microtubule-associated protein light chain 3B) is a core marker in the autophagy process, and its modification state directly reflects autophagy activity. The lipidation conversion of LC3B-I to LC3B-II promotes autophagosome formation and drives the degradation of damaged components; the continuous activation of LC3B-II can enhance the cell clearance ability, maintain homeostasis and resist external stress. In the prior art, the dynamics of LC3B are confirmed to provide a basis for the treatment of diseases targeting autophagy regulation and drug research and development.

[0184] It can be seen from the above results that compared with the control group (NC), the combination (Rib + AE + FDP) increased the expression level of LC3B more significantly than the single components of Rib, AE, and FDP, indicating that the combination of Rib + AE + FDP produced a synergistic effect that is not possessed by the single use of Rib, AE, and FDP.

[0185] Related tests were also carried out on other examples, and it was found that their effects were basically similar to those of the composition in Example 3.

[0186] Test Example 7

[0187] In this test example, the anti-wrinkle effects of Example 3 and the single-component substances with corresponding contents were tested. The specific experimental steps were as follows:

[0188] After resuscitating human dermal fibroblasts, they were cultured routinely. When the cell confluence rate reached about 60%, the cells were seeded into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0189] When the cell confluence rate in the 6-well plates reached about 60%, grouping and drug administration were carried out. Experimental grouping: The seeded cells were divided into a normal control group (NC), a model control group (MC), and a sample group. Among them, the normal control group (NC) and the model control group (MC) were not treated with anything and were continued to be cultured with fresh medium. The sample group was respectively added with a medium containing the multi-component composite cosmetic composition in Example 3, equal amounts of ribose, equal amounts of Artemia extract, or equal amounts of sodium fructose diphosphate, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0190] Among them, the addition amount of each substance was: calculated by the mass of the medium, the proportion of the corresponding substance was:

[0191] Rib+AE+FDP group (i.e., Example 3): 0.05% ribose, 0.03% Artemia extract, and 0.002% sodium fructose diphosphate;

[0192] Rib group: 0.05% ribose; AE group: 0.03% Artemia extract; FDP group: 0.002% sodium fructose diphosphate.

[0193] After the incubation was completed, the medium was discarded, and the cells were gently rinsed 1-2 times with D-Hanks balanced salt solution (D-HBSS). Then the model control group (MC) and the sample group were exposed to ultraviolet light for 50 s (UVA, 30 mJ / cm 2 )

[0194] After the model establishment was completed, the normal control group (NC) and the model control group (MC) were not treated with anything and were continued to be cultured with fresh medium for 24 h. The sample group was respectively added with the above-mentioned medium containing the multi-component composite cosmetic composition in Example 3, equal amounts of ribose, equal amounts of Artemia extract, or equal amounts of sodium fructose diphosphate, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0195] After the incubation was completed, the cells were fixed with 4% paraformaldehyde. After 30 min of fixation, immunofluorescence detection was carried out, photographed and observed under a microscope, and pictures were collected and analyzed.

[0196] The results are shown in Table 8 and Figure 8 as follows.

[0197] Table 8 Collagen content in each group

[0198] Sample Group Test Result SD P-Value Enhancement Rate (vs MC) Normal Control (NC) 1.00 0.120 0.0018** 120％ Model Control (MC) 0.45 0.045 / / Rib 0.74 0.102 0.0109* 64％ AE 0.69 0.060 0.0050** 53％ FDP 0.63 0.104 0.0506 40％ Rib + AE + FDP 0.85 0.121 0.0059** 88％

[0199] Among them, compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; three biological replicates (N = 3).

[0200] Type I collagen is the core component maintaining the firmness and elasticity of skin structure. Ultraviolet (UV) rays, oxidative stress or aging can activate matrix metalloproteinases (MMPs), degrade Collagen I and disrupt its reticular arrangement, resulting in skin laxity and wrinkle formation. The decrease in the content of Collagen I will directly affect the skin barrier function, elasticity and anti-wrinkle ability, accelerating photoaging and wrinkle generation. In this test example, the fluorescence intensity and distribution of Collagen I in skin tissues were analyzed by immunofluorescence quantification to evaluate whether the sample has an anti-wrinkle effect of promoting collagen synthesis or inhibiting degradation.

[0201] As can be seen from the above results, compared with the control group (MC), the composition (Rib + AE + FDP) increased the content of type I collagen more significantly than the single components of Rib, AE, and FDP, indicating that the combination of Rib + AE + FDP produced a synergistic effect that the single use of Rib, AE, and FDP did not have.

[0202] Related tests were also conducted on other embodiments, and it was found that their effects were basically similar to those of the composition in Example 3.

[0203] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and shall be included in the protection scope of the present invention.

Claims

1. A composition, characterized in that, The composition comprises: ribose, artemia extract and trisodium fructose diphosphate.

2. The composition according to claim 1, characterized in that, In the composition, the mass ratio of ribose, artemia extract and trisodium fructose diphosphate is 0.01 - 0.5:0.01 - 0.2:0.001 - 0.01; Preferably, in the composition, the mass ratio of ribose, artemia extract and trisodium fructose diphosphate is 0.03 - 0.3:0.01 - 0.15:0.001 - 0.

01.

3. The composition according to claim 1 or 2, characterized in that, The composition further comprises excipients acceptable in cosmetics; Preferably, the excipients acceptable in the cosmetics include: solvents, emulsifiers, stabilizers, thickeners, preservatives, fragrances, pigments, fillers and humectants.

4. The composition according to claim 3, wherein The composition further comprises other active ingredients for cosmetics; The active ingredients for cosmetics refer to substances having at least one of the following functions (1)-(6): (1) Promote the generation of skin barrier or enhance the skin barrier defense; (2) Moisturize; (3) Anti-wrinkle; (4) Tighten the skin; (5) Antioxidant; (6) Protect mitochondria or improve mitochondrial function.

5. The preparation method of the composition according to any one of claims 1 - 4, comprising the following steps: Mix ribose, artemia extract and trisodium fructose diphosphate according to the mass ratio described in claim 2, and that is obtained.

6. A cosmetic, characterized in that, The cosmetics contain the composition according to any one of claims 1 - 4; Preferably, in the cosmetics, calculated by the total mass of the cosmetics, the proportion of the composition is 0.6 - 5%.

7. The application of the composition according to any one of claims 1 - 4 in the preparation of cosmetics or drugs.

8. The application according to claim 7, characterized in that, The cosmetics or drugs have at least one of the following functions (1)-(6): (1) Promote the generation of skin barrier or enhance the skin barrier defense; (2) Moisturize; (3) Anti-wrinkle; (4) Tighten the skin; (5) Antioxidant; (6) Protect mitochondria or improve mitochondrial function.

9. The application according to claim 7, characterized in that In the cosmetics or drugs, calculated by the total mass, the proportion of the composition is 0.6 - 5%.

10. The application according to claim 7, characterized in that, The dosage forms of the cosmetics include: emulsions, aqueous solutions, oils, gels and powders; the dosage forms of the drugs include: ointments, aerosols and patches.

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