Cosmetic composition with multiple cell energy enhancement effects and application thereof

Abstract

The application discloses a cosmetic composition with multiple cell energy promotion effects and application thereof. The cosmetic composition in the application is obtained by compounding ribose, halocyprid extract and trisodium d-fructofuranose 1, 2, 3, 4-tetraacetate, and has significant synergistic effects among components, can produce multiple effects such as antioxidation, protection of mitochondria, promotion of skin barrier generation or enhancement of skin barrier defense, moisturization, anti-wrinkle, skin tightening and the like after compounding, thereby realizing a truly meaningful 'all-in-one', and meeting the use requirements of various types of consumer groups.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and in particular to a cosmetic composition with multiple cellular energy-boosting effects and its application. Background Technology

[0002] Traditional cosmetic compositions have long been limited by their singular efficacy, stemming from the lack of synergistic effects from the simple layering of active ingredients. Existing cosmetics often achieve basic efficacy coverage through the mechanical mixing of single-function ingredients. However, the differences in the physicochemical properties of different components can easily lead to activity loss or antagonistic interactions, resulting in the claimed "multi-effect combination" only providing localized effects. For example, heat-sensitive ingredients like vitamin C and retinol are difficult to balance in conventional formulations in terms of stability and transdermal efficiency, leading to insufficient duration of efficacy. Furthermore, existing technologies struggle to overcome the compatibility barriers between water-soluble (e.g., hyaluronic acid) and fat-soluble (e.g., squalane) active ingredients, making it impossible to simultaneously achieve multiple functions such as anti-oxidation, repair, and anti-aging. Although the industry has attempted to develop multifunctional ingredients through synthetic biotechnology (e.g., recombinant collagen) or the combination of natural active ingredients (e.g., Ganoderma lucidum extract), these approaches still face bottlenecks such as an imperfect efficacy evaluation system and soaring industrialization costs. Of course, there are also attempts to use multiple emulsion carriers to achieve precise regional encapsulation and phased 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 formulas, which greatly limits its market penetration.

[0003] Therefore, there is an urgent need to develop a simple cosmetic composition with multiple functions, so that it can be used as a functional combination in various cosmetics to achieve a true "multi-effect in one" effect. Summary of the Invention

[0004] This invention aims to at least solve one of the aforementioned technical problems existing in the prior art. Therefore, the object of this invention is to provide a cosmetic composition with multiple functions and its application. The composition of this invention achieves multiple functions through the combined action of ribose, artemia extract, and trisodium fructose diphosphate, and the components exhibit good synergistic effects, producing significant effects in promoting skin barrier formation or enhancing skin barrier defense; moisturizing; anti-wrinkle; skin firming; antioxidant; and protecting or improving mitochondrial function.

[0005] In a first aspect, the present invention provides a composition comprising: ribose, artichoke extract, and trisodium fructose diphosphate.

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

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

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

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

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

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

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

[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 moisturizing agent includes, but is not limited to, glycerin and hyaluronic acid;

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

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

[0017] In some embodiments of the present invention, the active ingredient for cosmetics refers to a substance having at least one of the following functions (1)-(6):

[0018] (1) Promotes skin barrier formation or enhances skin barrier defense;

[0019] (2)Moisturizing;

[0020] (3) Anti-wrinkle;

[0021] (4) Tightens skin;

[0022] (5) Antioxidant;

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

[0024] A second aspect of the present invention provides a method for preparing the composition described above, comprising the following steps:

[0025] The ribose, artichoke extract, and trisodium fructose diphosphate are mixed according to the mass ratios described above to obtain the final product.

[0026] A third aspect of the present invention provides a cosmetic product comprising the composition described in the above aspects.

[0027] In some embodiments of the present invention, the composition accounts for 0.6-5% of the total mass of the cosmetic.

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

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

[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 moisturizing agent includes, but is not limited to, glycerin and hyaluronic acid;

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

[0033] A fourth aspect of the invention provides the use of the compositions described in the above aspects in the preparation of cosmetics or pharmaceuticals.

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

[0035] (1) Promotes skin barrier formation or enhances skin barrier defense;

[0036] (2)Moisturizing;

[0037] (3) Anti-wrinkle;

[0038] (4) Tightens skin;

[0039] (5) Antioxidant;

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

[0041] In some embodiments of the present invention, the composition comprises 0.6-5% of the total mass of the cosmetic or pharmaceutical product.

[0042] In some embodiments of the present invention, the cosmetic or pharmaceutical product is a topical preparation.

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

[0044] In some embodiments of the present invention, the dosage form of the medicine includes: ointment, aerosol, and patch.

[0045] The beneficial effects of this invention are:

[0046] This invention provides a cosmetic composition with multiple functions, in which the components have a significant synergistic effect, and can simultaneously produce multiple functions such as anti-oxidation; protection of mitochondria; promotion of skin barrier formation or enhancement of skin barrier defense; moisturizing; anti-wrinkle; and skin firming, thereby achieving a true "multi-effect in one" and meeting the usage needs of various types of consumers. Attached Figure Description

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

[0048] Figure 2 The ratio of red to green average fluorescence intensity of mitochondrial membrane potential after treatment in different experimental groups.

[0049] Figure 3 The relative expression levels of the FLG gene after treatment in different experimental groups.

[0050] Figure 4 NAD after treatment in different experimental groups + / NADH ratio.

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

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

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

[0054] Figure 8 The collagen content after treatment in different experimental groups. Detailed Implementation

[0055] The present invention will be further described in detail below through specific embodiments. Unless otherwise specified, the raw materials, reagents, or apparatus used in the embodiments and comparative examples are all available from conventional commercial sources or can be obtained by existing technical methods. Unless otherwise specified, the test or experimental methods are conventional methods in the art.

[0056] In the following embodiments, ribose, Artemia extract, and trisodium 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 this invention.

[0057] Example 1

[0058] This embodiment provides a cosmetic composition with multiple functions, the components of which are:

[0059] By weight, 0.03 parts ribose, 0.018 parts Artemia extract, and 0.0012 parts trisodium fructose diphosphate.

[0060] The preparation method is as follows:

[0061] According to the above-mentioned mass ratio, sugar, Artemia extract, and trisodium fructose diphosphate are mixed evenly to obtain a cosmetic composition with multiple functions.

[0062] Example 2

[0063] This embodiment provides a cosmetic composition with multiple functions, the components of which are:

[0064] By weight, 0.25 parts ribose, 0.15 parts Artemia extract, and 0.01 parts trisodium fructose diphosphate.

[0065] The preparation method is as follows:

[0066] According to the above-mentioned mass ratio, sugar, Artemia extract, and trisodium fructose diphosphate are mixed evenly to obtain a cosmetic composition with multiple functions.

[0067] Example 3

[0068] This embodiment provides a cosmetic composition with multiple functions, the components of which are:

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

[0070] The preparation method is as follows:

[0071] According to the above-mentioned mass ratio, sugar, Artemia extract, and trisodium fructose diphosphate are mixed evenly to obtain a cosmetic composition with multiple functions.

[0072] Test Example 1

[0073] In this test case, the energy enhancement effect of Example 1 and the corresponding content of the single-component substance was tested. The specific experimental steps are as follows:

[0074] After resuscitating human immortalized epidermal cells (HaCaT), they were cultured routinely until the cell plating rate reached about 60%. The cells were then seeded into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0075] When the cell seeding rate in the 6-well plates reached approximately 60%, the cells were divided into groups for drug administration. The cells were divided into a normal control group (NC) and a sample group. The normal control group (NC) received no treatment and continued culturing in fresh culture medium. The sample group was incubated with culture medium containing the multi-component cosmetic composition from Example 1, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and incubated at 37°C and 5% CO2 for 24 hours.

[0076] The amounts of each substance added are as follows: based on the mass of the culture medium, the percentage of each substance is as follows:

[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 changed. The normal control group (NC) was not treated and continued to be cultured for 24 hours using fresh culture medium. The sample groups were respectively added to the culture medium containing the multi-component cosmetic composition of Example 1, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and continued to be incubated for 24 hours at 37°C and 5% CO2.

[0080] After incubation, the ATP content was detected using a commercially available kit.

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

[0082] Table 1 Energy Enhancement Results for 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 universal molecule that directly provides energy within cells, playing a central role in energy metabolism and physiological function regulation. When the body's energy demand increases, ATP releases high-energy phosphate bonds through hydrolysis, providing immediate energy support. Studies have shown that the rate of ATP synthesis and its reserve levels are directly related to cell viability and tissue fatigue resistance. By detecting ATP levels, an individual's energy metabolism efficiency can be assessed, providing a basis for energy enhancement strategies.

[0087] The results above show that, compared with the control group (NC), the combination (Rib+AE+FDP) significantly increased ATP content more than Rib, AE, and FDP alone, indicating that the combination of Rib+AE+FDP produced a synergistic effect that Rib, AE, and FDP alone could not achieve.

[0088] Other embodiments were also tested and found to have effects that were basically similar to those of the composition in Example 1.

[0089] Test Example 2

[0090] In this test case, the mitochondrial activity improvement effect of Example 2 and the corresponding concentrations of 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 plating rate reached about 60%, the cells were seeded into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0092] When the cell seeding rate in the 6-well plates reached approximately 60%, the cells were divided into groups for drug administration. The cells were divided into three groups: a normal control group (NC), a model control group (MC), and a sample group. The NC and MC groups received no treatment and were cultured in fresh culture medium. The sample group was incubated with culture medium containing the multi-component cosmetic composition from Example 2, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and incubated at 37°C and 5% CO2 for 24 hours.

[0093] The amounts of each substance added are as follows: based on the mass of the culture medium, the percentage of each substance is as follows:

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

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

[0096] After incubation, discard the culture medium and gently rinse the cells 1-2 times with D-Hanks balanced salt solution (D-HBSS). Then, expose the model control group (MC) and the sample group to UV light for 50 seconds (UVA, 9 mJ / cm²). 2 ).

[0097] After modeling, the normal control group (NC) and the model control group (MC) were not treated and were cultured for another 24 hours using fresh culture medium. The sample groups were respectively added to the culture medium containing the multi-component cosmetic composition of Example 2, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and were incubated for another 24 hours at 37°C and 5% CO2.

[0098] After incubation, JC-1 staining was performed, and the samples were observed and photographed under a fluorescence microscope.

[0099] The fluorescence intensity (S) of each group was analyzed using ImageJ, and the average red / green fluorescence intensity ratio of the mitochondrial membrane potential was calculated (the calculation formula is shown below).

[0100]

[0101] In the formula:

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

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

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

[0105] Sample group Test results SD p-value Improvement rate (vs NC) Normal control (NC) 1.24 0.028 <0.0001**** 86％ Model Comparison (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] 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. Decreased MMP weakens ATP synthesis, leading to ROS accumulation and mitochondrial dysfunction; conversely, increased MMP enhances electron transport chain activity, promotes efficient energy production, reduces oxidative damage, and restores mitochondrial homeostasis, thereby directly improving mitochondrial vitality. Its levels can be detected using fluorescent probes, providing a basis for assessing mitochondrial health and intervening in energy metabolism issues.

[0108] The results above show that, compared with the control group (MC), the combination (Rib+AE+FDP) significantly enhances the mitochondrial membrane potential, which is more significant than that of Rib, AE, and FDP alone. This indicates that the combination of Rib+AE+FDP produces a synergistic effect that cannot be achieved by using Rib, AE, and FDP alone.

[0109] Other embodiments were also tested and found to have effects that were basically similar to those of the composition in Example 2.

[0110] Test Example 3

[0111] In this test case, the barrier repair effect of Example 3 and the corresponding concentration of the single-component substance was tested. The specific experimental steps are as follows:

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

[0113] When the cell seeding rate in the 6-well plates reached approximately 60%, the cells were divided into groups for drug administration. The cells were divided into a normal control group (NC) and a sample group. The normal control group (NC) received no treatment and was cultured in fresh culture medium. The sample group was incubated with culture medium containing the multi-component cosmetic composition from Example 3, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and incubated at 37°C and 5% CO2 for 24 hours.

[0114] The amounts of each substance added are as follows: based on the mass of the culture medium, the percentage of each substance is as follows:

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

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

[0117] After incubation, the culture medium was replaced. The normal control group (NC) was not treated and continued to be cultured for 24 hours using fresh culture medium. The sample groups were respectively added to the culture medium containing the multi-component cosmetic composition of Example 3, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and continued to be incubated for 24 hours at 37°C and 5% CO2.

[0118] After incubation, total RNA was extracted using a commercially available kit, reverse transcribed, and the relative expression levels of β-actin (internal reference) and FLG genes were detected using q-PCR.

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

[0120] Table 3 Barrier repair results for each group

[0121] Sample group Test results SD p-value Improvement 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 (figrin) is a core protein of the skin barrier function, and its expression level directly determines the integrity of the barrier. FLG deficiency or degradation leads to a loose stratum corneum structure, reduced moisturizing factor (NMF), and increased moisture loss, causing inflammation and sensitivity. Conversely, enhanced FLG synthesis promotes stratum corneum densification, improves moisturizing capacity, and blocks external stimuli, thereby accelerating barrier repair. FLG status can be assessed through immunohistochemistry or genetic testing, providing a basis for targeted skincare to repair the skin barrier.

[0124] The results above show that, compared with the control group (NC), the combination (Rib+AE+FDP) significantly increased the relative expression level of the FLG gene, which is more significant than that of Rib, AE, and FDP alone. This indicates that the combination of Rib+AE+FDP produced a synergistic effect that Rib, AE, and FDP alone could not achieve.

[0125] Other embodiments were also tested and found to have effects that were basically similar to those of the composition in Example 3.

[0126] Test Example 4

[0127] In this test case, the effects of Example 1 and the corresponding amounts of the single-component substance 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 seeded into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0129] When the cell seeding rate in the 6-well plates reached approximately 60%, the cells were divided into groups for drug administration. The cells were divided into three groups: a normal control group (NC), a model control group (MC), and a sample group. The NC and MC groups received no treatment and were cultured in fresh culture medium. The sample group was incubated with culture medium containing the multi-component cosmetic composition from Example 1, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and incubated at 37°C and 5% CO2 for 24 hours.

[0130] The amounts of each substance added are as follows: based on the mass of the culture medium, the percentage of each substance is as follows:

[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% trisodium fructose diphosphate.

[0133] After incubation, discard the culture medium and gently rinse the cells 1-2 times with D-Hanks balanced salt solution (D-HBSS). Then, expose the model control group (MC) and the sample group to UV light for 50 seconds (UVA, 9 mJ / cm²). 2 ).

[0134] After modeling, the normal control group (NC) and the model control group (MC) were not treated and were cultured for another 24 hours using fresh culture medium. The sample groups were respectively added to the culture medium containing the multi-component cosmetic composition of Example 1, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and were incubated for another 24 hours at 37°C and 5% CO2.

[0135] After incubation, NAD was detected using a commercially available kit. + And NADH, and calculate NAD + / NADH ratio.

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

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

[0138] Sample group Test results SD p-value Improvement Rate (vs. NC) Normal control (NC) 11.72 0.875 0.0012** 64％ Model Comparison (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] NAD + The NAD / NADH ratio affects a cell's ability to produce energy. In the cytoplasm, NAD+ is required to convert glucose into pyruvate via glycolysis. + In mitochondria, the TCA cycle reduces NAD. + The molecules generate multiple NADH molecules, which are then oxidized by complex I of the electron transport chain (ETC) to produce ATP. + Decreased levels can affect mitochondrial function, overall cellular health, and the development of age-related diseases. This test case detects NAD+. + The NADH / NADH ratio level was used to assess whether the sample had an effect on improving mitochondrial dysfunction.

[0141] The results above show that, compared with the control group (MC), the combination (Rib+AE+FDP) significantly increased the NAD+ / NADH content, indicating that the combination of Rib+AE+FDP produced a synergistic effect that Rib, AE, and FDP alone could not achieve.

[0142] Other embodiments were also tested and found to have effects that were basically similar to those of the composition in Example 1.

[0143] Test Example 5

[0144] In this test case, the moisturizing efficacy of Example 2 and the corresponding amounts of single-component substances 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 into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0146] When the cell seeding rate in the 6-well plates reached approximately 60%, the cells were divided into groups for drug administration. The cells were divided into a normal control group (NC) and a sample group. The normal control group (NC) received no treatment and was cultured in fresh culture medium. The sample group was incubated with culture medium containing the multi-component cosmetic composition from Example 2, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and incubated at 37°C and 5% CO2 for 24 hours.

[0147] The amounts of each substance added are as follows: based on the mass of the culture medium, the percentage of each substance is as follows:

[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 incubation, the normal control group (NC) was not treated and continued to be cultured for 24 hours using fresh culture medium. The sample groups were respectively added to the culture medium containing the multi-component cosmetic composition of Example 2, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and continued to be incubated for 24 hours at 37°C and 5% CO2.

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

[0152]

[0153] In the formula:

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

[0155] Simultaneously, total RNA was extracted from each experimental group, cDNA was synthesized, and q-PCR was used to detect the gene expression of β-actin and the target gene. β-actin was used as an internal control 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 Tables 5 and 6. Figure 5 , Figure 6 As shown.

[0160] Table 5 HA content of each group

[0161]

[0162]

[0163] 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 levels of the AQP3 gene

[0165] Sample group Test results SD p-value Upward adjustment 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] Compared with NC, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; three biological replicates (N=3).

[0167] The skin's moisturizing system, from the inside out, can be summarized as follows: water capture, water retention, water activation, and water storage. Hyaluronic acid (HA) in the dermis absorbs a large amount of water, acting as the skin's water reservoir, providing hydration for the entire skin layer, and also filling the dermis to make its structure more complete. Aquaporins (AQPs) are widely distributed in various tissues of organisms, influencing the body's water metabolism. AQPs are highly expressed in the keratinocyte membranes of the epidermal basal and spinous layers, acting as water and glycerol transport proteins in these membranes, promoting skin hydration, and are a key factor in maintaining skin hydration. Therefore, this test case evaluates the moisturizing efficacy of the sample by detecting the hyaluronic acid (HA) content and the relative expression level of the AQP3 gene.

[0168] The results above show that, compared with the control group (NC), the moisturizing effect of the combination (Rib+AE+FDP) is more significant than that of Rib, AE, and FDP alone, indicating that the combination of Rib+AE+FDP produces a synergistic effect that Rib, AE, and FDP alone cannot achieve.

[0169] Other embodiments were also tested and found to have effects that were basically similar to those of the composition in Example 2.

[0170] Test Example 6

[0171] In this test case, the autophagy-promoting effect of Example 3 and the corresponding amounts of single-component substances were tested. The specific experimental steps are as follows:

[0172] After thawing, the fibroblasts were cultured routinely. When the cell plating rate reached about 60%, the cells were seeded 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 approximately 60%, the cells were divided into groups for drug administration. The cells were divided into a normal control group (NC) and a sample group. The normal control group (NC) received no treatment and was cultured in fresh culture medium. The sample group was incubated with culture medium containing the multi-component cosmetic composition from Example 3, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and incubated at 37°C and 5% CO2 for 24 hours.

[0174] The amounts of each substance added are as follows: based on the mass of the culture medium, the percentage of each substance is as follows:

[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% trisodium fructose diphosphate.

[0177] After incubation, the culture medium was replaced. The normal control group (NC) was not treated and continued to be cultured for 24 hours using fresh culture medium. The sample groups were respectively added to the culture medium containing the multi-component cosmetic composition of Example 3, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and continued to be incubated for 24 hours at 37°C and 5% CO2.

[0178] After incubation, RNA was extracted using a commercially available kit, reverse transcribed into cDNA, and then detected by quantitative real-time PCR.

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

[0180] Table 7 Autophagy enhancement effects of each group

[0181] Sample group Test results SD p-value Improvement 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 biomarker of autophagy, and its modification state directly reflects autophagic activity. The lipidation conversion of LC3B-I to LC3B-II promotes autophagosome formation and drives the degradation of damaged components; sustained activation of LC3B-II enhances cellular clearance capacity, maintains homeostasis, and resists external stress. Current technologies utilize the dynamics of LC3B to provide a basis for targeted regulation of autophagy in disease treatment and drug development.

[0184] The results above show that, compared with the control group (NC), the combination (Rib+AE+FDP) significantly increased LC3B expression levels, indicating that the combination of Rib+AE+FDP produced a synergistic effect that Rib, AE, and FDP alone could not achieve.

[0185] Other embodiments were also tested and found to have effects that were basically similar to those of the composition in Example 3.

[0186] Test Example 7

[0187] In this test case, the anti-wrinkle effect of Example 3 and the corresponding amount of single-component substance was tested. The specific experimental steps are as follows:

[0188] After resuscitating human dermal fibroblasts, they were cultured routinely. When the cell plating 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 seeding rate in the 6-well plates reached approximately 60%, the cells were divided into groups for drug administration. The cells were divided into three groups: a normal control group (NC), a model control group (MC), and a sample group. The NC and MC groups received no treatment and were cultured in fresh culture medium. The sample group was incubated with culture medium containing the multi-component cosmetic composition from Example 3, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and incubated at 37°C and 5% CO2 for 24 hours.

[0190] The amounts of each substance added are as follows: based on the mass of the culture medium, the percentage of each substance is as follows:

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

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

[0193] After incubation, discard the culture medium and gently rinse the cells 1-2 times with D-Hanks balanced salt solution (D-HBSS). Then, expose the model control group (MC) and the sample group to UV light for 50 seconds (UVA, 30 mJ / cm²). 2 ).

[0194] After modeling, the normal control group (NC) and the model control group (MC) were not treated and were cultured for another 24 hours using fresh culture medium. The sample groups were respectively added to the culture medium containing the multi-component cosmetic composition of Example 3, an equal amount of ribose, an equal amount of Artemia extract, or an equal amount of trisodium fructose diphosphate, and were incubated for another 24 hours at 37°C and 5% CO2.

[0195] After incubation, cells were fixed with 4% paraformaldehyde for 30 minutes. Immunofluorescence was then performed, and images were taken and analyzed under a microscope.

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

[0197] Table 8 shows the collagen content of each group.

[0198] Sample group Test results SD P-Value Improvement Rate (vs. MC) Normal control (NC) 1.00 0.120 0.0018** 120％ Model Comparison (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 a core component for maintaining skin firmness and elasticity. Ultraviolet (UV) radiation, oxidative stress, or aging can activate matrix metalloproteinases (MMPs), degrading Collagen I and disrupting its reticular arrangement, leading to skin laxity and wrinkle formation. Decreased Collagen I levels directly affect skin barrier function, elasticity, and anti-wrinkle ability, accelerating photoaging and wrinkle formation. This test case uses immunofluorescence quantitative analysis of the fluorescence intensity and distribution of Collagen I in skin tissue to assess whether the sample has an anti-wrinkle effect by promoting collagen synthesis or inhibiting degradation.

[0201] The results above show that, compared with the control group (MC), the combination (Rib+AE+FDP) significantly increased the content of type I collagen, which is more significant than that of Rib, AE, and FDP alone. This indicates that the combination of Rib+AE+FDP produced a synergistic effect that Rib, AE, and FDP alone could not achieve.

[0202] Other embodiments were also tested and found to have effects that 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 changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. Application of the composition in the preparation of cosmetics or pharmaceuticals; The composition comprises ribose, artichoke extract, and trisodium fructose diphosphate. 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; The cosmetic or pharmaceutical product has the following functions (1)-(3): (1) Promotes skin barrier formation or enhances skin barrier defense; (2) Moisturizing; (3) Protect mitochondria or improve mitochondrial function.

2. The application according to claim 1, characterized in that, In the cosmetic or pharmaceutical product, the composition accounts for 0.6-5% of the total mass.

3. The application according to claim 1, characterized in that, The dosage forms of the cosmetics include: emulsions, aqueous solutions, oils, gels, and powders; the dosage forms of the pharmaceuticals include: ointments, aerosols, and patches.

4. The application according to claim 1, characterized in that, The cosmetics or pharmaceuticals mentioned also include excipients acceptable for cosmetic use.

5. The application according to claim 4, characterized in that, Acceptable excipients in the cosmetics include: solvents, emulsifiers, stabilizers, thickeners, preservatives, fragrances, pigments, fillers, and moisturizers.

6. The application according to claim 1, characterized in that, The cosmetics or pharmaceuticals also include other active ingredients used in cosmetics; The active ingredient used in cosmetics refers to a substance having at least one of the following functions: (1)-(6) (1) Promotes skin barrier formation or enhances skin barrier defense; (2) Moisturizing; (3) Anti-wrinkle; (4) Tightens skin; (5) Antioxidant; (6) Protect mitochondria or improve mitochondrial function.

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