Skin anti-aging substance composition with mitochondrial enhancing effect, preparation method, application and product thereof

Abstract

The invention belongs to the technical field of biological medicines and cosmetics, and particularly relates to a skin anti-aging substance composition with a mitochondrial enhancing effect, a preparation method, application and a product thereof. The composition consists of spermidine, quercetin, a mirabilis jalapa extract, vitamin K2, adenosine and a solvent. The composition provided by the invention can repair a mitochondrial ultrastructure, recover ATP generation and maintain mitochondrial membrane potential, and can also reduce accumulation of an aging marker SA-beta-gal, up-regulate type I collagen and enhance Sirt3 expression.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine and cosmetics, and specifically relates to a skin anti-aging substance composition with a mitochondrion enhancing effect, a preparation method, application and product thereof. BACKGROUND

[0002] Mitochondria, as the main organelle, is closely related to intrinsic and extrinsic skin aging. Skin tissue with high metabolic activity depends on mitochondrial ATP generation to support the rapid proliferation and renewal of its cells. Notably, age-related mitochondrial dysfunction manifests through a series of phenotypic changes, including mitochondrial network fragmentation, loss of membrane potential and accumulation of mitochondrial DNA mutations, which are considered as markers of skin aging. These structural and functional changes impair energy metabolism while exacerbating the production of reactive oxygen species (ROS), forming a vicious cycle that accelerates the aging of dermal fibroblasts and epidermal keratinocytes. Especially in photoaged skin, ultraviolet (UV) radiation directly damages mitochondrial components, further amplifying oxidative stress and inflammatory response, leading to the degradation of extracellular matrix (ECM) integrity. Therefore, regulating mitochondrial function by enhancing mitochondrial homeostasis through bioactive compounds has become an important strategy to delay skin aging.

[0003] Although enhancing mitochondrial homeostasis is considered a key strategy to delay skin aging, the current application of traditional mitochondrial modulators has many shortcomings. These modulators often have low efficiency and high production cost, and are limited to a single target in detection, making it difficult to comprehensively regulate mitochondrial function, which to some extent limits their wide application and effect improvement in the field of anti-skin aging.

[0004] Patent CN101579355B provides a nutritional composition for maintaining mitochondrial function, which is composed of coenzyme Q10, selenium element and free radical scavenger. The composition provided by the patent has a complementary and enhanced effect in maintaining mitochondrial function compared with any single component, can improve the activity of "respiratory chain" to transfer electrons to manufacture ATP, and protect mitochondrial DNA from damage. The medicine or health care product made of the composition as the core component can be used for delaying aging, preventing the occurrence of senile degenerative diseases, and treating cardiovascular and cerebrovascular diseases. Patent CN112423726B provides a composition composed of effective amounts of nicotinamide, quercetin or its derivative rutin, zinc or its salt, ascorbic acid, parsley or its derivative apigenin, and alpha-lipoic acid. It is suitable for enhancing the tolerance of cells to DNA damage, oxidative stress and mitochondrial dysfunction, or improving DNA repair capacity.

[0005] Developing more efficient and comprehensive mitochondrial modulators has become an urgent need in the field of anti-skin aging research. SUMMARY

[0006] In view of the above, the present application provides a skin anti-aging substance composition with a mitochondria-enhancing effect, a preparation method, application and product thereof.

[0007] The technical solution of the present application is as follows: In one aspect, the present application provides a skin anti-aging substance composition with a mitochondria-enhancing effect, which is composed of spermidine, quercetin, Mirabilis jalapa extract, vitamin K2, adenosine and a solvent.

[0008] Specifically, the skin anti-aging substance composition comprises 0.1-3% w / v spermidine, 0.001-0.01% w / v quercetin, 0.01-1% w / v Mirabilis jalapa extract, 0.003-0.1% w / v vitamin K2 and 0.1-1% w / v adenosine.

[0009] Preferably, the skin anti-aging substance composition comprises 0.1% w / v spermidine, 0.001% w / v quercetin, 1% w / v Mirabilis jalapa extract, 0.1% w / v vitamin K2 and 1% w / v adenosine.

[0010] Further preferably, the skin anti-aging substance composition comprises 3% w / v spermidine, 0.01% w / v quercetin, 0.01% w / v Mirabilis jalapa extract, 0.003% w / v vitamin K2 and 0.1% w / v adenosine.

[0011] Specifically, the solvent comprises any one or more of water, polyhydric alcohol solvents, alcohol solvents, oil solvents, but is not limited thereto.

[0012] Preferably, the solvent is water.

[0013] In another aspect, the present application provides a preparation method of the skin anti-aging substance composition.

[0014] Specifically, the preparation method comprises mixing spermidine, quercetin, Mirabilis jalapa extract, vitamin K2 and adenosine in proportion, and then adding them into water to obtain the composition.

[0015] In another aspect, the present application provides a composition preparation comprising the skin anti-aging substance composition.

[0016] Specifically, the composition preparation further comprises any one or more of plant extracts, vitamin substances, bioactive peptide substances, enzyme substances, acid substances, polysaccharide substances or mineral elements.

[0017] Preferably, the plant extracts include, but are not limited to, any one or more of the following: ginseng extract, grape seed extract, licorice extract, green tea extract, aloe vera extract, witch hazel extract, rosemary extract, centella asiatica extract, peony extract, angelica extract, coix seed extract, poria cocos extract, polygonum cuspidatum extract, mulberry bark extract, honeysuckle extract, orange peel extract, lemon extract, peach blossom extract, ginkgo leaf extract, wolfberry extract, rose extract, lavender extract, chamomile extract, atractylodes macrocephala extract, peony root bark extract, scutellaria baicalensis extract, astragalus membranaceus extract, angelica dahurica extract, bletilla striata extract, oat extract, eucommia ulmoides extract, kudzu root extract, polygonatum odoratum extract, rehmannia glutinosa extract, eleutherococcus senticosus extract, cornus officinalis extract, salvia miltiorrhiza extract, chrysanthemum extract, cinnamon extract, magnolia officinalis extract, schisandra chinensis extract, amomum villosum extract, and cistanche deserticola extract.

[0018] Specifically, the vitamins mentioned include fat-soluble vitamins and / or water-soluble vitamins.

[0019] Preferably, the fat-soluble vitamins include, but are not limited to, any one or more of vitamin A, vitamin D, vitamin E, or vitamin K; Preferably, the water-soluble microorganisms include, but are not limited to, any one or more of the following: vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B12, vitamin C, choline, or inositol.

[0020] Preferably, the bioactive peptides include, but are not limited to, any one or more of the following: collagen peptides, elastin peptides, glutathione, casein phosphopeptides, hyaluronic acid hydrolysate peptides, ceramide peptides, soybean peptides, corn peptides, sea cucumber peptides, oyster peptides, bird's nest peptides, carnosine, or lactoferrin peptides.

[0021] Preferably, the enzymes include, but are not limited to, any one or more of the following: superoxide dismutase, catalase, glutathione peroxidase, bromelain, papain, and lysozyme.

[0022] Preferably, the acidic substances include, but are not limited to, any one or more of the following: vitamin A and its derivatives, vitamin C and its derivatives, fruit acids, salicylic acid, arbutin, tranexamic acid, kojic acid, ferulic acid, or azelaic acid.

[0023] Specifically, the polysaccharides include, but are not limited to, any one or more of the following: fungal polysaccharides, animal polysaccharides, plant polysaccharides, and algal polysaccharides.

[0024] Preferably, the fungal polysaccharide includes, but is not limited to, any one or more of Pachyman, Tremella polysaccharide, Flammulina velutipes polysaccharide, Auricularia auricula polysaccharide, Cordyceps polysaccharide, Chuan Huang polysaccharide, Cloud polysaccharide, or any one or more of the following: polysaccharide.

[0025] Preferably, the animal polysaccharide includes any one or more of glycogen, chondroitin sulfate, hyaluronic acid, heparin, chitosan, chitin, or glycosaminoglycan; Preferably, the plant polysaccharide includes any one or more of cellulose, hemicellulose, pectin, inulin, konjac polysaccharide, astragalus polysaccharide, polysaccharide, polysaccharide, polysaccharide, polysaccharide, or polysaccharide. Preferably, the algal polysaccharide includes any one or more of green algal polysaccharide, red algal polysaccharide, or brown algal polysaccharide.

[0026] Preferably, the mineral element includes, but is not limited to, any one or more of zinc, copper, iron, magnesium, calcium, or silicon.

[0027] In another aspect, the present application provides the use of the skin anti-aging material composition or composition preparation of any one of the above in the preparation of a product, wherein the product is selected from any one of the following products: (1) cosmetics with anti-inflammatory, whitening, anti-wrinkle, tightening or repair skin barrier function; (2) drugs with anti-inflammatory, whitening, anti-wrinkle, tightening or repair skin barrier function; (3) health products that help antioxidant.

[0028] In another aspect, the present application provides a cosmetic product, which comprises the skin anti-aging material composition or composition preparation of any one of the above.

[0029] Specifically, the cosmetic product includes any one or more of the following: cream, mask, eye cream, neck cream, hand cream, body milk, moisturizing lotion, essence, toner, shampoo, hair conditioner, shower gel or cleansing milk.

[0030] Specifically, the cosmetic product further comprises a cosmetically acceptable excipient.

[0031] Preferably, the cosmetically acceptable excipient includes one or more of humectants, thickeners, emollients, skin conditioners, emulsifiers, preservatives, emollients, lubricants, stabilizers.

[0032] In another aspect, the present application provides a drug, which comprises the skin anti-aging material composition or composition preparation of any one of the above.

[0033] Specifically, the drug further comprises a pharmaceutically acceptable excipient.

[0034] Preferably, the pharmaceutically acceptable excipient includes, but is not limited to, a solvent, a diluent, a disintegrant, a precipitation inhibitor, a surfactant, a glidant, a binder, a lubricant, a dispersant, a suspending agent, an isotonic agent, a thickening agent, an emulsifying agent, a preservative, a stabilizer, a hydrating agent, an emulsification accelerator, a buffer, an absorbent, a coloring agent, a flavoring agent, a sweetening agent, an ion exchanger, a release agent, a coating agent, a flavoring agent, or an antioxidant.

[0035] Specifically, the dosage form of the pharmaceutical product includes, but is not limited to, a gastrointestinal administration dosage form and a non-gastrointestinal administration dosage form, according to the administration mode.

[0036] Preferably, the gastrointestinal administration dosage form includes, but is not limited to, a tablet, a powder, a granule, a solution, a capsule, an emulsion, a suspension, an oil.

[0037] Preferably, the gastrointestinal administration dosage form includes, but is not limited to, an injection administration dosage form, a respiratory administration dosage form, a skin administration dosage form, a mucosa administration dosage form, and a cavity administration dosage form.

[0038] In another aspect, the present application provides a health product, which comprises the skin anti-aging substance composition or the composition preparation according to any one of the above.

[0039] Specifically, the dosage form of the health product includes a solid dosage form, a semi-solid dosage form, or a liquid dosage form.

[0040] Specifically, the health product further comprises a nutritionally acceptable nutritional additive.

[0041] Preferably, the nutritional additive includes, but is not limited to, any one or more of dietary fiber, prebiotic, protein, lipid substance, mineral, and vitamin.

[0042] The present application has the following beneficial effects: The skin anti-aging substance composition provided by the present application can repair the ultrastructure of mitochondria, restore ATP generation, maintain the membrane potential of mitochondria, and also reduce the accumulation of aging marker SA-β-gal, up-regulate collagen type I, and enhance Sirt3 expression. BRIEF DESCRIPTION OF DRAWINGS

[0043] Figure 1 Representative images of the mitochondrial network dyed by MitoTracker Red; the blue in the figure is the Hoechst channel, and the red is the MitoTracker channel, and the scale bar is 100 microns.

[0044] Figure 2To restore the mitochondrial morphological-functional integrity of UVB-irradiated HDF cells to CC-1 and CC-2; Figure A shows the clustering heatmap analysis of mitochondrial morphometric parameters; Figure B shows the quantitative assessment of mitochondrial morphological characteristics: fluorescence intensity and density; the fluorescence intensity and density are characterized by mitochondrial parameters and are unitless; in the figure, * represents p<0.05 vs model group, **** represents p<0.0001 vs model group; #### represents p<0.0001 vs control group; % represents p<0.05 vs CC-1 group, %% represents p<0.01 vs CC-1 group, %%%% represents p<0.0001 vs CC-1 group; $$$$ represents p<0.0001 vs CC-2 group.

[0045] Figure 3 The effect of CC-1 and CC-2 on ATP levels in a UVB-damaged HDF cell model; * in the figure represents p<0.05 vs model group; ** represents p<0.01 vs model group; ## represents p<0.01 vs control group.

[0046] Figure 4 The results of JC-1 detection of mitochondrial membrane potential in HDF cells are shown in the fluorescence image (red: J-aggregates; green: monomers) with a scale bar of 100 micrometers; and the red-green fluorescence ratio is used for quantification. In the figure, ** represents p<0.01 vs. model group; ## represents p<0.01 vs. control group.

[0047] Figure 5 The image shows the phenotypes associated with the mitigation of aging in human dermal fibroblasts by CC-1 and CC-2. In the figure, A represents the representative images of SA-β-Gal staining in each group of HDF cells; B shows the quantitative analysis results of SA-β-Gal activity; and C shows the results of Western blot detection of aging markers (type I collagen and Sirt3). Detailed Implementation

[0048] The present invention will be further clearly and completely illustrated below through examples. These examples are only a part of the embodiments of the present invention and are not intended to limit the invention, but are merely illustrative. Unless otherwise specified, the experimental methods used in the following examples are routine experiments. Unless otherwise specified, the materials and reagents used in the following examples are commercially available. The Mirabilis jalapa extract of the present invention was purchased from NAOLYS, batch number OE [MJ+C] / 16 (S-Gly).

[0049] Data in this invention are expressed as mean ± standard deviation. One-way ANOVA was used for comparisons among multiple groups, processed using GraphPad Prism software. P < 0.05 was considered statistically significant.

[0050] Example 1: Preparation of the skin anti-aging substance composition CC-1 The skin anti-aging composition of Example 1 is CC-1, which contains: 0.1% w / v spermidine, 0.001% w / v quercetin, 1% w / v Mirabilis jalapa extract, 0.1% w / v vitamin K2 and 1% w / v adenosine.

[0051] The preparation method of the skin anti-aging substance composition in Example 1 is as follows: Weigh out 1 g spermidine, 0.01 g quercetin, 10 g Mirabilis jalapa extract, 1 g vitamin K2 and 10 g adenosine respectively, mix well and add water to make up to 1000 mL.

[0052] Example 2: Preparation of the skin anti-aging substance composition CC-2 The skin anti-aging composition of Example 2 is CC-2, which contains: 3% w / v spermidine, 0.01% w / v quercetin, 0.01% w / v Mirabilis jalapa extract, 0.003% w / v vitamin K2 and 0.1% w / v adenosine.

[0053] The preparation method of the skin anti-aging substance composition in Example 2 is as follows: Weigh out 30 g spermidine, 0.1 g quercetin, 0.1 g Mirabilis jalapa extract, 0.03 g vitamin K2 and 1 g adenosine respectively, mix well and add water to make up to 1000 mL.

[0054] Experimental Example 1: Mitochondrial Morphology Measurement Dermal fibroblasts (HDFs) were seeded at a rate of 3000 cells / well in 96-well plates for 24 hours. 1:200 (dilution factor) of CC-1 from Example 1, or 1:200 (dilution factor) of CC-2 from Example 2, or spermidine solution (0.2 mg / mL), or quercetin solution (0.2 mg / mL), or Mirabilis jalapa extract solution (0.2 mg / mL), or vitamin K2 solution (0.2 mg / mL), or adenosine solution (0.2 mg / mL) were added respectively. HDFs cells treated with basal culture medium served as the model group, and were pretreated for 24 hours.

[0055] After pretreatment, it was subjected to UVB (1.1 J / cm). 2 Irradiation (power 1.1 mW / cm) 2Cells were irradiated with UVB for 1000 s, with HDFs cells treated with basal culture medium but not exposed to UVB serving as the control group. Cells were then incubated at 37 °C in a 5% CO2 incubator for 24 hours. Fluorescent probes Mito-Tracker Red CMXRos and Hoechst 33342 were added, and the cells were incubated at 37 °C for 30 minutes. The fluorescence of Mito-Tracker Red CMXRos reflecting mitochondrial morphology was detected. Images were captured using an ImageXpress® Confocal HT.ai system, and morphological feature parameters were extracted automatically in batches using the image analysis method developed by Werner JH Koopman's team (Multiplexed High-Content Analysis of Mitochondrial Morphofunction Using Live-Cell Microscopy. NatProtoc 2016, 11, 1693–1710, doi:10.1038 / nprot.2016.094.) via Image Pro Plus 6.0 software.

[0056] Mitochondria are key organelles that maintain functional integrity through a dynamic balance of fusion and fission events. UVB-exposed HDFs exhibit severe mitochondrial structural perturbations, including volume shrinkage, network disintegration, and perinuclear aggregation. Figure 1 Both CC-1 and CC-2 treatments effectively protected the ultrastructure of mitochondria. Treated cells primarily exhibited short rod-shaped mitochondria, with restored network-like structure, evenly distributed in the cytoplasm. Quantitative morphological analysis showed significant recovery of parameters such as fluorescence intensity and density, and other measured parameters also showed a trend towards normalization compared to the control group. Figure 2 ).

[0057] Experimental Example 2: CC-1 and CC-2 enhance ATP production and ΔΨm stability 1. ATP production measurement HDF cells were seeded at a rate of 3000 cells / well in 96-well plates for 24 hours. Then, either CC-1 (Example 1) diluted 1:200 or CC-2 (Example 2) diluted 1:200 were added to the cells, using the basal culture medium-treated HDF cells as the model group, and the cells were pretreated for 24 hours.

[0058] After pretreatment, it was subjected to UVB (1.1 J / cm). 2 Irradiation (power 1.1 mW / cm) 2Cells were irradiated for 1000 s, with HDF cells that had not received UVB irradiation and were treated with basal culture medium serving as the control group. Cells were then incubated at 37 °C in a 5% CO2 incubator for 24 hours. Cell ATP production was measured using the CellTiter-Glo luminescence assay (Promega).

[0059] 2. Measurement of mitochondrial membrane potential (ΔΨm) HDF cells were seeded at a rate of 3000 cells / well in 96-well plates for 24 hours. Then, either CC-1 (Example 1) diluted 1:200 or CC-2 (Example 2) diluted 1:200 were added to the cells, using the basal culture medium-treated HDF cells as the model group, and the cells were pretreated for 24 hours.

[0060] After pretreatment, it was subjected to UVB (1.1 J / cm). 2 Irradiation (power 1.1 mW / cm) 2 Cells were irradiated for 1000 s, with HDFs cells that had not received UVB irradiation and were treated with basal culture medium serving as the control group. Cells were then incubated at 37 °C in a 5% CO2 incubator for 24 hours. Mitochondrial membrane potential (ΔΨm) was measured using the JC-1 fluorescent probe. Relative ΔΨm was expressed as the ratio of red (J-aggregates, 525 / 590 nm) to green (monomers, 490 / 530 nm) fluorescence intensity; a higher ratio indicated a higher degree of polarization. Images were captured and analyzed using the ImageXpress® Confocal HT.ai system.

[0061] 3. Experimental Results In the HDF cell model of UVB damage, both CC-1 and CC-2 significantly increased ATP levels under UVB stress, further demonstrating their mitochondrial protective potential. Figure 3 ).

[0062] Figure 4 UVB irradiation induced a significant reduction in ΔΨm (red-green ratio decreased by 30%, p<0.01 vs control group). CC-1 and CC-2 treatments significantly attenuated this effect, maintaining ΔΨm at 118% and 97.7% of the control group, respectively (p<0.01 vs UVB group).

[0063] Experimental Example 3: Effects of CC-1 and CC-2 on the protein expression levels of SA-β-gal, type I collagen, and Sirt3 HDF cells were seeded at a rate of 300,000 cells / well in 6-well plates for 24 hours. HDF cells treated with basal culture medium were added at a 1:200 dilution of CC-1 from Example 1 or CC-2 from Example 2 at a 1:200 dilution, respectively, as the model group, and pretreated for 24 hours.

[0064] After pretreatment, it was subjected to UVB (1.1 J / cm). 2 Irradiation (power 1.1 mW / cm) 2 The cells were irradiated for 1000 s, with HDFs cells that had not received UVB irradiation and were treated with basal culture medium as the control group. They were then incubated at 37 ℃ in a 5% CO2 incubator for 24 hours.

[0065] β-galactosidase (SA-β-gal) staining was performed using a kit (Cell signaling technology, catalog number #9860); the protein expression levels of type I collagen and Sirt3 were detected by Western blotting. Results showed that both CC-1 and CC-2 significantly reduced the accumulation of SA-β-gal in human dermal fibroblasts (HDFs) after UVB irradiation. Figure 5 (A and B in the original text). Compared with the UVB exposure group alone, CC-1 and CC-2 treatments upregulated the expression of type I collagen, a key structural protein in skin aging; notably, CC-1 also enhanced the expression of Sirt3, a longevity-related protein, suggesting additional anti-aging benefits. Figure 5 (C in the middle).

[0066] The above detailed description is a specific illustration of one feasible embodiment of the present invention, and this embodiment is not intended to limit the patent scope of the present invention. It should be noted that all equivalent implementations or modifications made without departing from the present invention should be included within the scope of the technical solution of the present invention. Therefore, the protection scope of the present invention should be determined by the appended claims.

Claims

1. A skin anti-aging substance composition with mitochondrial-enhancing effects, characterized in that, The skin anti-aging substance composition consists of spermidine, quercetin, four o'clock extract, vitamin K2, adenosine, and a solvent.

2. The skin anti-aging substance composition according to claim 1, characterized in that, The skin anti-aging substance composition comprises: 0.1-3% w / v spermidine, 0.001-0.01% w / v quercetin, 0.01-1% w / v Mirabilis jalapa extract, 0.003-0.1% w / v vitamin K2 and 0.1-1% w / v adenosine.

3. The skin anti-aging substance composition according to claim 1, characterized in that, The solvents include any one or more of the following: water, polyol solvents, alcohol solvents, and oil solvents.

4. A method for preparing the skin anti-aging substance composition according to any one of claims 1-3, characterized in that, The preparation method includes: mixing spermidine, quercetin, four o'clock extract, vitamin K2 and adenosine in a certain proportion, and then adding them to water to obtain a skin anti-aging substance composition.

5. A compositional formulation comprising the skin anti-aging substance composition according to any one of claims 1-4.

6. The composition formulation according to claim 5, characterized in that, The composition formulation further includes any one or more of the following: plant extracts, vitamins, bioactive peptides, enzymes, acids, polysaccharides, or mineral elements.

7. The use of the skin anti-aging substance composition according to any one of claims 1-3 or the composition formulation according to any one of claims 5-6 in the preparation of a product, characterized in that, The product mentioned is selected from any one of the following products: (1) Cosmetics with anti-inflammatory, whitening, anti-wrinkle, firming or skin barrier repair effects; (2) Medicines with anti-inflammatory, whitening, anti-wrinkle, firming or skin barrier repair effects; (3) Health products that help fight oxidation.

8. A cosmetic product, characterized in that, The cosmetic product comprises the skin anti-aging substance composition according to any one of claims 1-4 or the composition formulation according to any one of claims 6-7.

9. A medicine, characterized in that, The medicine comprises the skin anti-aging substance composition according to any one of claims 1-4 or the composition formulation according to any one of claims 6-7.

10. A health product, characterized in that, The health product comprises the skin anti-aging substance composition according to any one of claims 1-4 or the composition formulation according to any one of claims 6-7.

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