Anti-aging composition and application thereof

Abstract

The invention provides an anti-aging composition and application thereof, the composition at least contains pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or salt thereof in physiologically acceptable media, the composition contains PQQ, ergothioneine and 5-aminolevulinic acid (5-ALA) or salt thereof, the three components are compounded to achieve a synergistic effect, and the anti-aging effect is achieved. The dyeing intensity of beta-galactosidase is reduced, and the anti-aging effect is achieved.

Description

Technical Field

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

[0002] Aging is the process of functional and organic decline and aging of the body during aging, including aging damage at different levels of tissues and organs, genes, proteins, cells, and intercellular communication. It is specifically manifested as telomere shortening, accumulation of DNA damage, abnormal oncogene activation, metabolic changes, and the massive production of reactive oxygen species (ROS). Among them, telomeres, as nuclear protein structures covering the ends of each chromosome arm, are particularly susceptible to damage caused by aging and oxidative stress. When telomeres continue to shorten due to cell division, it will aggravate the apoptosis and death of cells in the body, eventually leading to aging of the body; the production of telomerase can slow down the telomere shortening process, thereby delaying aging.

[0003] Abnormal expression of specific enzymes is closely linked to the biogenesis and degradation pathways of organelles in aging cells. β-galactosidase, a lysosomal enzyme that catalyzes the hydrolysis of glycosidic bonds, has been identified as overexpressed and significantly accumulated in most aging cells. This enzyme activity, found in many normal cells under physiological conditions, is significantly amplified in aging cells due to increased lysosomal content, making it a biomarker of cellular aging. When protein levels are damaged, the homeostasis of protein synthesis, folding, and degradation is disrupted, leading to elevated levels of the biomarker enzyme β-galactosidase and accelerating aging and the onset of age-related diseases. At the cellular level, manifestations include stem cell damage, mitochondrial dysfunction, and nutritional imbalances. Stem cell damage leads to decreased tissue repair capacity, resulting in irreversible aging, while mitochondrial dysfunction and nutritional imbalances accelerate aging.

[0004] The aging process of zebrafish bears a strong resemblance to that of humans, both manifesting as accumulation of lipofuscin and cognitive decline. Studies have shown that drug-induced zebrafish aging models also exhibit similar DNA damage, cell tissue damage, and organ damage. When H₂O₂ enters the zebrafish's body, it generates a large number of free radicals. Excessive free radicals trigger oxidative stress and lipid peroxidation, which destabilizes biomembranes, produces lipofuscin, damages nuclear and mitochondrial DNA, causes oxidative protein damage, and cross-links macromolecules, thereby inducing aging and shortening the fish's lifespan.

[0005] According to the body's aging mechanism, improving mitochondrial activity and reducing oxygen free radicals can effectively improve the adverse effects of aging. Summary of the Invention

[0006] The present application provides a more effective composition for slowing down aging, which comprises pyrroloquinoline quinone disodium salt (PQQ), ergothioneine and 5-aminolevulinic acid or its salt. The three substances can synergistically enhance their effectiveness, thereby slowing down aging and can be used to reduce the occurrence of age-related diseases.

[0007] The specific technical solutions of this application are as follows:

[0008] 1. A composition comprising at least pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or a salt thereof in a physiologically acceptable medium.

[0009] Furthermore, the 5-aminolevulinic acid or its salt is not particularly limited, but is preferably a pharmaceutically acceptable inorganic or organic acid addition salt. Inorganic acid addition salts include hydrochlorides, hydrobromides, sulfates, nitrates, and phosphates. Organic acid addition salts include acetates, lactates, citrates, tartrates, succinates, maleates, fumarates, ascorbates, and the like, preferably 5-aminolevulinic acid phosphate and / or hydrochloride.

[0010] 2. The composition of claim 1 , wherein the mass ratio of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or a salt thereof is 1:(0.5-15):(0.5-2.0).

[0011] 3. The composition of claim 1 , wherein the mass ratio of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or a salt thereof is 1:(0.5-15):(0.75-1.5).

[0012] 4. The composition according to any one of items 1 to 3, wherein the dosage form of the composition comprises at least one or more of tablets, powders, granules, capsules or aqueous solutions.

[0013] 5. Use of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt in the preparation of anti-aging products.

[0014] Furthermore, the 5-aminolevulinic acid or its salt is not particularly limited, but is preferably a pharmaceutically acceptable inorganic or organic acid addition salt. Inorganic acid addition salts include hydrochlorides, hydrobromides, sulfates, nitrates, and phosphates. Organic acid addition salts include acetates, lactates, citrates, tartrates, succinates, maleates, fumarates, ascorbates, and the like, preferably 5-aminolevulinic acid phosphate and / or hydrochloride.

[0015] 6. The method according to claim 5, wherein the mass ratio of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or a salt thereof is 1:(0.5-15):(0.5-2.0), preferably 1:(0.5-15):(0.75-1.5).

[0016] 7. Use of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt in the preparation of a product for inhibiting or reducing β-galactosidase.

[0017] Furthermore, the 5-aminolevulinic acid or its salt is not particularly limited, but is preferably a pharmaceutically acceptable inorganic or organic acid addition salt. Inorganic acid addition salts include hydrochlorides, hydrobromides, sulfates, nitrates, and phosphates. Organic acid addition salts include acetates, lactates, citrates, tartrates, succinates, maleates, fumarates, ascorbates, and the like, preferably 5-aminolevulinic acid phosphate and / or hydrochloride.

[0018] 8. The method of claim 7, wherein the mass ratio of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or a salt thereof is 1:(0.5-15):(0.5-2.0), preferably 1:(0.5-15):(0.75-1.5).

[0019] Effects of the Invention

[0020] (1) The composition provided in the present application contains PQQ, ergothioneine and 5-aminolevulinic acid (5-ALA) or its salt. The three are combined to synergistically enhance the effect, reduce the staining intensity of β-galactosidase, and achieve an anti-aging effect.

[0021] (2) All raw materials used in this application are of natural fermentation origin and are highly safe.

[0022] (3) The present invention has low production cost, short process cycle, low pollution, low energy consumption, and is easy to industrialize. DETAILED DESCRIPTION

[0023] The present application is described in detail below in conjunction with the described embodiments. Although specific embodiments of the present application are shown, it should be understood that the present application can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to enable a more thorough understanding of the present application and to fully convey the scope of the present application to those skilled in the art.

[0024] It should be noted that certain words are used in the specification and claims to refer to specific components. Those skilled in the art should understand that technicians may use different nouns to refer to the same component. This specification and claims do not use the difference in nouns as a way to distinguish components, but use the difference in the functions of the components as the criterion for distinction. As mentioned throughout the specification and claims, "including" or "comprising" are open-ended terms and should be interpreted as "including but not limited to". The subsequent description of the specification is a preferred embodiment of the present application, but the description is based on the general principles of the specification and is not intended to limit the scope of the present application. The scope of protection of this application shall be as defined by the attached claims.

[0025] The present application provides a composition comprising at least pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or a salt thereof in a physiologically acceptable medium.

[0026] In the present application, there is no particular limitation on the 5-aminolevulinic acid salt, which is a salt conventionally used in the art, preferably a pharmaceutically acceptable acid addition salt of an inorganic acid or organic acid, for example, an inorganic acid addition salt includes hydrochloride, hydrobromide, sulfate, nitrate and phosphate. Organic acid addition salts include acetate, lactate, citrate, tartrate, succinate, maleate, fumarate, ascorbate, etc., preferably 5-aminolevulinic acid phosphate and / or hydrochloride.

[0027] "Physiologically acceptable medium" refers to a medium suitable for topical application in contact with mammalian skin, particularly human skin.

[0028] The composition described in the present application contains pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt. The three can synergistically enhance the effect, slow down aging, and can be used to reduce the occurrence of age-related diseases.

[0029] In some embodiments, the mass ratio of the pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt is 1:(0.5-15):(0.5-2.0), preferably 1:(0.5-15):(0.75-1.5).

[0030] In the present application, there is no particular limitation on the 5-aminolevulinic acid salt, which is a salt conventionally used in the art, preferably a pharmaceutically acceptable acid addition salt of an inorganic acid or organic acid. For example, inorganic acid addition salts include hydrochlorides, hydrobromides, sulfates, nitrates, and phosphates. Organic acid addition salts include acetates, lactates, citrates, tartrates, succinates, maleates, fumarates, ascorbic acid salts, and the like, preferably 5-aminolevulinic acid phosphate and / or hydrochloride.

[0031] For example, the mass ratio of the pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt (m 吡咯并喹啉醌二钠盐 :m 麦角硫因 :m 5-氨基乙酰丙酸 ) can be 1:0.5:0.5, 1:0.5:0.75, 1:0.5:1, 1:0.5:1.25, 1:0.5:1.5, 1:0.5:1.75, 1:0.5:2, 1:1:0.75, 1:1.5:0.75, 1:2:0.75, 1:2.5:0.75, 1:3:0.75, 1:3.5:0.75, 1:4:0.75, 1:4.5:0.75, 1:5:0.75, 1:5.5:0.75, 1:6:0.75, 1:6.5 :0.75, 1:7:0.75, 1:7.5:0.75, 1:8:0.75, 1:8.5:0.75, 1:9:0.75, 1:9.5:0.75, 1:10:0.75, 1:10.5:0.75, 1:11:0.75, 1:11.5:0.75, 1:12:0.75, 1:12.5:0.75, 1:13:0.75, 1:13.5:0.75, 1:14:0.75, 1:14.5:0.75, 1:15:0.75, etc.

[0032] In some embodiments, the composition further contains other active ingredients and / or excipients acceptable in the food or cosmetic field.

[0033] In this application, there is no limitation on other active ingredients, and those skilled in the art can make routine selections based on actual needs. Similarly, there is no limitation on the content of other active ingredients in the composition, and those skilled in the art can choose the amount of other active ingredients to be added based on actual needs.

[0034] In this application, there is no limitation on the excipients, and those skilled in the art can select conventional excipients in the art based on actual needs. Similarly, there is no limitation on the content of the excipients, and those skilled in the art can select the amount of the excipients based on actual needs.

[0035] In some embodiments, the dosage form of the composition includes at least one or more of tablets, powders, granules, capsules or aqueous solutions.

[0036] In the present application, there is no limitation on the preparation methods of tablets, powders, granules, capsules and aqueous solutions, which can be prepared by conventional methods in the art and by selecting appropriate excipients.

[0037] In the present application, there is no restriction on the content of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt in the composition, as long as the ratio of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt meets the technical effects described in the present application.

[0038] The present application provides the use of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt in the preparation of anti-aging products.

[0039] The pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt synergistically enhance the effect and can significantly reduce the staining intensity of β-galactosidase, so that the pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt can be applied to anti-aging and used to prepare anti-aging products.

[0040] In some embodiments, the mass ratio of the pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt is 1:(0.5-15):(0.5-2.0), preferably 1:(0.5-15):(0.75-1.5).

[0041] As mentioned above, in the present application, there is no particular limitation on the 5-aminolevulinic acid salt, which is a salt conventionally used in the art, preferably a pharmaceutically acceptable acid addition salt of an inorganic acid or organic acid, for example, inorganic acid addition salts include hydrochloride, hydrobromide, sulfate, nitrate and phosphate. Organic acid addition salts include acetate, lactate, citrate, tartrate, succinate, maleate, fumarate, ascorbate, etc., preferably 5-aminolevulinic acid phosphate and / or hydrochloride.

[0042] For example, the mass ratio of the pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt (m 吡咯并喹啉醌二钠盐 :m 麦角硫因 :m 5-氨基乙酰丙酸) can be 1:0.5:0.5, 1:0.5:0.75, 1:0.5:1, 1:0.5:1.25, 1:0.5:1.5, 1:0.5:1.75, 1:0.5:2, 1:1:0.75, 1:1.5:0.75, 1:2:0.75, 1:2.5:0.75, 1:3:0.75, 1:3.5:0.75, 1:4:0.75, 1:4.5:0.75, 1:5:0.75, 1:5.5:0.75, 1:6:0.75, 1:6.5 :0.75, 1:7:0.75, 1:7.5:0.75, 1:8:0.75, 1:8.5:0.75, 1:9:0.75, 1:9.5:0.75, 1:10:0.75, 1:10.5:0.75, 1:11:0.75, 1:11.5:0.75, 1:12:0.75, 1:12.5:0.75, 1:13:0.75, 1:13.5:0.75, 1:14:0.75, 1:14.5:0.75, 1:15:0.75, etc.

[0043] The present application provides the use of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt in the preparation of a product for inhibiting or reducing β-galactosidase.

[0044] As described above, pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt can synergistically enhance efficacy and significantly reduce the staining intensity of β-galactosidase, and can be used to prepare products that inhibit or reduce β-galactosidase.

[0045] In some embodiments, the mass ratio of the pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt is 1:(0.5-15):(0.5-2.0), preferably 1:(0.5-15):(0.75-1.5).

[0046] As mentioned above, there is no particular limitation on the 5-aminolevulinic acid salt, and it is a salt conventionally used in the art, preferably a pharmaceutically acceptable acid addition salt of an inorganic acid or organic acid. For example, inorganic acid addition salts include hydrochlorides, hydrobromides, sulfates, nitrates, and phosphates. Organic acid addition salts include acetates, lactates, citrates, tartrates, succinates, maleates, fumarates, ascorbic acid salts, and the like, preferably 5-aminolevulinic acid phosphate and / or hydrochloride.

[0047] For example, the mass ratio of the pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt (m 吡咯并喹啉醌二钠盐 :m 麦角硫因 :m 5-氨基乙酰丙酸) can be 1:0.5:0.5, 1:0.5:0.75, 1:0.5:1, 1:0.5:1.25, 1:0.5:1.5, 1:0.5:1.75, 1:0.5:2, 1:1:0.75, 1:1.5:0.75, 1:2:0.75, 1:2.5:0.75, 1:3:0.75, 1:3.5:0.75, 1:4:0.75, 1:4.5:0.75, 1:5:0.75, 1:5.5:0.75, 1:6:0.75, 1:6.5 :0.75, 1:7:0.75, 1:7.5:0.75, 1:8:0.75, 1:8.5:0.75, 1:9:0.75, 1:9.5:0.75, 1:10:0.75, 1:10.5:0.75, 1:11:0.75, 1:11.5:0.75, 1:12:0.75, 1:12.5:0.75, 1:13:0.75, 1:13.5:0.75, 1:14:0.75, 1:14.5:0.75, 1:15:0.75, etc.

[0048] In this application, water-soluble is a commonly used administration method for zebrafish research in the art, which refers to dissolving the compound to be tested (such as a drug, toxin or nutrient) directly in the water environment where the zebrafish lives, and allowing the compound to enter the fish body through the skin, gills or food through contact with water.

[0049] Example

[0050] The present application provides a general and / or specific description of the materials and test methods used in the experiments. In the following examples, unless otherwise specified, % represents wt%, i.e., weight percentage. The reagents or instruments used without indicating the manufacturer are all conventional reagent products that can be obtained commercially, among which pyrroloquinoline quinone disodium salt (PQQ), ergothioneine MitoEGT TM , 5-aminolevulinic acid phosphate (5-ALA phosphate) were both from Bloomage Biotech Co., Ltd.

[0051] Example 1 Preparation of tablets

[0052] (1) Mixing: 5-aminolevulinic acid phosphate (5-ALA phosphate), ergothioneine, pyrroloquinoline quinone disodium salt (PQQ) and auxiliary materials are uniformly mixed; wherein the mass ratio of PQQ, ergothioneine and 5-ALA phosphate is 1:10:1, the total mass of 5-aminolevulinic acid phosphate (5-ALA phosphate), ergothioneine and pyrroloquinoline quinone disodium salt is 42g, and the auxiliary materials are composed of 29g of starch and 29g of sodium carboxymethyl cellulose;

[0053] (2) Wet granulation: After mixing evenly, 10 ml of purified water was added by spraying for wet granulation;

[0054] (3) Drying: The wet granulated material was placed in an oven and dried at a constant temperature of 50°C for 4 h. After drying, it was sterilized with ultraviolet light for 15 min to obtain whole granules.

[0055] (4) Tablet compression: The dried whole granules are tablet compressed to obtain 80 tablets, each weighing 1.0 g.

[0056] Example 2 Preparation of granules

[0057] (1) Mixing: PQQ, ergothioneine and 5-aminolevulinic acid phosphate (5-ALA phosphate) are uniformly mixed in a mass ratio of 1:5:1.5; wherein the total mass of 5-ALA phosphate, ergothioneine and pyrroloquinoline quinone disodium salt PQQ is 1 kg;

[0058] (2) One-step granulation: After uniform mixing, the mixture was added to an FBM-3 boiling granulator (equipment capacity: 3 L). A 30 wt% aqueous solution containing ethanol was used as a wetting agent and sprayed from the top of the boiling granulator. The atomization pressure of the boiling granulator was 0.1 MPa and the inlet air temperature was 60°C. After processing for 84 min, the mixture was screened using 20-mesh and 60-mesh sieves to obtain granules with a mesh size of 60 to 80.

[0059] (3) Sterilization: The material obtained in step (2) is sterilized by ultraviolet light for 30 min;

[0060] (4) Packaging: The sterilized material was packaged to obtain 180 bags of granules, with each bag containing 5 g.

[0061] Example 3 Preparation of powder

[0062] Seven different powders were prepared, with specific formulations shown in Table 1, as samples required for subsequent efficacy tests.

[0063] (1) Mixing: PQQ, ergothioneine and 5-ALA phosphate were mixed, the total mass of each group of active ingredients was 100 g, and the addition ratio of each group of active ingredients is shown in Table 1;

[0064] (2) Sieving: Sieve the mixed material through a 60-mesh Taylor standard sieve to obtain a uniform powdered material;

[0065] (3) Sterilization: The material obtained in step (2) is sterilized by ultraviolet light for 30 min;

[0066] (4) Packaging: The sterilized material was packaged to obtain 32 bags of powder, each bag containing 2.5 g of powder, which were Examples 3-1 to 3-7, respectively.

[0067] Preparation of Comparative Examples 1-3 Powder

[0068] Comparative Examples 1-3 are all single-functional ingredient powders, each containing PQQ, ergothioneine and 5-ALA phosphate, which are sterilized by ultraviolet light for 30 minutes and then packaged, with a packing amount of 2.5 g per bag.

[0069] Preparation of Comparative Examples 4-6 Powders

[0070] (1) Mixing: PQQ and thioneine were mixed, the total mass of the functional ingredients was 100 g, and the proportion of functional ingredients added was shown in Table 1;

[0071] (2) Sieving: The mixed material is passed through a 60-mesh Taylor standard sieve to obtain a uniform powdered material;

[0072] (3) Sterilization: The material obtained in step (2) is sterilized by ultraviolet light for 30 min;

[0073] (4) Packaging: The sterilized material was packaged to obtain 32 bags of powder, with each bag containing 2.5 g.

[0074] Preparation of Comparative Examples 7-11 Powder

[0075] (1) Mixing: PQQ and 5-ALA phosphate were mixed, the total mass of the active ingredients was 100 g, and the proportion of active ingredients added was shown in Table 1;

[0076] (2) Sieving: The mixed material is passed through a 60-mesh Taylor standard sieve to obtain a uniform powdered material;

[0077] (3) Sterilization: The material obtained in step (2) is sterilized by ultraviolet light for 30 min;

[0078] (4) Packaging: The sterilized material was packaged to obtain 32 bags of powder, with each bag containing 2.5 g.

[0079] Preparation of Comparative Examples 12-17 Powders

[0080] (1) Mixing: thioneine was mixed with 5-ALA phosphate, the total mass of the functional ingredients was 100 g, and the functional ingredients were added in a proportion shown in Table 1;

[0081] (2) Sieving: The mixed material is passed through a 60-mesh Taylor standard sieve to obtain a uniform powdered material;

[0082] (3) Sterilization: The material obtained in step (2) is sterilized by ultraviolet light for 30 min;

[0083] (4) Packaging: The sterilized material was packaged to obtain 32 bags of powder, with each bag containing 2.5 g.

[0084] Table 1

[0085]

[0086]

[0087] Note: “-” indicates that the corresponding component is not added; “√” indicates that the corresponding component is added

[0088] Experimental Example 1 Anti-aging efficacy test

[0089] β-galactosidase (SA-β-gal) is an enzyme highly expressed in lysosomes, and its activity is significantly increased in aging cells. Its increased activity is closely related to aging characteristics such as cell cycle arrest, accumulation of DNA damage, and telomere shortening. The aging pathways of zebrafish and mammals (such as p53 / p21, NF-κB, mTOR, etc.) are highly conserved, and changes in SA-β-gal activity in zebrafish tissues (such as skin, brain, and liver) can be compared to the aging process of mammals. Multiple studies have confirmed that SA-β-gal activity increases with age in zebrafish aging models, while anti-aging treatments (such as rapamycin and resveratrol) can reduce its levels (such as Cell Reports, 2020). Similar results have also been widely reported in mouse and human cells, supporting its cross-species applicability.

[0090] 1. Experimental Methods

[0091] The powder prepared in Example 3 was used as the solute, and the solvent was standard dilution water.

[0092] Positive control: resveratrol, white powder, batch number F2121344, Shanghai Aladdin Biochemical Technology Co., Ltd., solvent is DMSO.

[0093] Zebrafish were kept in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to each liter of reverse osmosis water, conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3). The experimental animal use license number is: SYXK (Zhejiang) 2022-0004. The breeding and management complies with the requirements of the international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2025-10992-01.

[0094] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA), 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China).

[0095] Hydrogen peroxide solution (lot number G2023089, Shanghai Aladdin Biochemical Technology Co., Ltd., China); dimethyl sulfoxide (DMSO, lot number BCCD8942, Sigma, Switzerland); methylcellulose (lot number C2004046, Shanghai Aladdin Biochemical Technology Co., Ltd., China); 4% tissue cell fixative (lot number 20221014, Beijing Solebeau Technology Co., Ltd., China); cell senescence β-galactosidase staining kit (lot number 091821220526, Shanghai Beyotime Biotechnology Co., Ltd., China).

[0096] 6 hpf wild-type AB strain zebrafish were randomly selected and plated in 6-well plates. Thirty zebrafish were treated in each well. Samples (concentrations shown in Table 2) and a positive control, resveratrol at 20.0 μg / mL, were administered in water. A normal control group and a model control group were also established. The volume per well was 3 mL. All groups, except the normal control group, were treated with hydrogen peroxide in water to establish the zebrafish aging model. The medium was changed daily. After 6 days of treatment at 28°C, zebrafish in each experimental group were stained using the Cell Senescence β-Galactosidase Staining Kit (Lot No. 091821220526, Shanghai Beyotime Biotechnology Co., Ltd., China). After staining, 10 zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope.

[0097] 2. Evaluation Indicators

[0098] Data were collected using NIS-Elements D 3.20 advanced image processing software. The mean zebrafish β-galactosidase staining intensity was analyzed and statistically analyzed to evaluate the anti-aging efficacy of the samples. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software. A p < 0.05 indicated statistical significance.

[0099] 3. Experimental results and analysis. The results are shown in Table 2.

[0100] Table 2 Sample anti-aging efficacy evaluation test results (n=10)

[0101]

[0102]

[0103] Compared with the model control group (between the two groups), *p<0.05, **p<0.01, ***p<0.001

[0104] The staining intensity of β-galactosidase (β-gal) is often used as an aging-related marker (such as SA-β-gal, senescence-associated β-galactosidase). Changes in its intensity can indirectly reflect the effect of anti-aging substances. Stronger staining usually indicates more accumulation of senescent cells in the tissue, suggesting an accelerated aging process. Conversely, weakened staining may reflect that anti-aging substances have reduced senescent cells and have a better anti-aging effect.

[0105] Under the experimental conditions of this study, PQQ, ergothioneine, and 5-ALA all had anti-aging effects, specifically by reducing the average β-galactosidase staining intensity. As can be seen from Table 2 above, the average β-galactosidase staining intensity of each experimental group was lower than that of the model group, and the results were significantly different.

[0106] SPSS software was used to perform multiple analyses on the mean values ​​of β-galactosidase staining intensity in each group.

[0107] (1) Example 3-1 is compared with Comparative Examples 1 to 4, 7, and 12 groups respectively. The results are shown in Table 3. The results show that the numerical difference between the two groups is significant; the effect of PQQ, thioneine, and 5-ALA after composite in a ratio of 1:15:0.75 is better than that of using PQQ, thioneine, and 5-ALA alone and composite in pairs in equal proportions.

[0108] (2) Example 3-2 was compared with Comparative Examples 1 to 3, 5, 8, and 13, respectively. The results are shown in Table 4. The results show that the numerical differences between the two groups are all significant; Explanation: The effect of PQQ, ergothioneine, and 5-ALA phosphate after compounding in a ratio of 1:0.5:1.5 is better than using PQQ, ergothioneine, and 5-ALA phosphate alone and compounding in pairs in the same ratio.

[0109] (3) Example 3-3 was compared with Comparative Examples 1 to 3, 6, 9, and 14, respectively. The results are shown in Table 5. The results show that the numerical differences between the two groups are all significant; it can be seen that the effect of PQQ, ergothioneine, and 5-ALA phosphate after compounding in a 1:6:1 ratio is better than that of using PQQ, ergothioneine, 5-ALA phosphate alone and compounding in pairs in the same ratio.

[0110] (4) Example 3-4 was compared with Comparative Examples 1 to 3, 5, 10, and 15, respectively. The results are shown in Table 6. The results show that the numerical differences between the two groups are all significant; it shows that the effect of PQQ, ergothioneine and 5-ALA phosphate after compounding in a ratio of 1:0.5:0.5 is better than using pyrroloquinoline quinone disodium salt, ergothioneine and ergothioneine + 5-ALA phosphate and PQQ + 5-ALA phosphate compounded in pairs in the same proportion alone, but there is no significant difference compared with using 5-ALA phosphate alone and PQQ + ergothioneine compounded in the same proportion.

[0111] (5) Example 3-5 is compared with Comparative Examples 1 to 4, 11, and 16 groups, respectively. The results are shown in Table 7, and the results show that the numerical differences between the two groups are all significant. Explanation: The effect of PQQ, thioneine, and 5-ALA phosphate after compounding in a ratio of 1:15:2 is better than using PQQ, thioneine, and the same ratio compounded in pairs alone, but there is no significant difference compared with using 5-ALA phosphate alone.

[0112] (6) Examples 3-6 were compared with Comparative Examples 1 to 3, 5, 11, and 17, respectively. The results are shown in Table 8, and the results show that the numerical differences between the two groups are all significant; the effect of PQQ, ergothioneine, and 5-ALA phosphate after compounding in a ratio of 1:0.5:2 is better than using PQQ, ergothioneine, and the same ratio compounded in pairs, but there is no significant difference compared with using 5-ALA phosphate alone.

[0113] Table 3

[0114]

[0115]

[0116] Note: Compared with Example 3-1 (between two groups), *p<0.05, **p<0.01, ***p<0.001

[0117] Table 4

[0118]

[0119] Note: Compared with Example 3-2 (between two groups), *p<0.05, **p<0.01, ***p<0.001

[0120] Table 5

[0121]

[0122] Note: Compared with Example 3-3 (between two groups), *p<0.05, **p<0.01, ***p<0.001

[0123] Table 6

[0124]

[0125] Note: Compared with Example 3-4 (between two groups), *p<0.05, **p<0.01, ***p<0.001

[0126] Table 7

[0127]

[0128] Note: Compared with Examples 3-5 (between two groups), *p<0.05, **p<0.01, ***p<0.001

[0129] Table 8

[0130]

[0131] Note: Compared with Examples 3-6 (between two groups), *p<0.05, **p<0.01, ***p<0.001

[0132] It can be seen from the test results that in the composition described in the present application, when the mass ratio of PQQ, ergothioneine and 5-ALA phosphate is 1:0.5-15:0.5-2.0, especially 1:0.5-15:0.75-1.5, the β-galactosidase staining intensity can be significantly reduced, with better anti-aging effect.

[0133] In summary, the composition contains PQQ, ergothioneine and 5-aminolevulinic acid or its salt, which can synergistically enhance the effect, significantly reduce the β-galactosidase staining intensity, and has an excellent anti-aging effect.

[0134] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application in any other manner. Any person skilled in the art may utilize the above disclosed technical content to modify or modify the present application into equivalent embodiments with equivalent variations. However, any simple modifications, equivalent variations, and modifications to the above embodiments that do not depart from the technical content of the present application and are based on the technical essence of the present application shall still fall within the scope of protection of the present application.

Claims

1. A composition comprising at least pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or a salt thereof in a physiologically acceptable medium.

2. compositions according to claim 1, the mass ratio of wherein said pyrroloquinoline quinone disodium salt, thioneine and 5-aminolevulinic acid or its salt is 1:(0.5-15):(0.5-2.0).

3. compositions according to claim 1, the mass ratio of wherein said pyrroloquinoline quinone disodium salt, thioneine and 5-aminolevulinic acid or its salt is 1:(0.5-15):(0.75-1.5).

4. The composition according to any one of claims 1 to 3, wherein The dosage form of the composition includes at least one or more of tablets, powders, granules, capsules or aqueous solutions.

5. Use of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt in the preparation of anti-aging products.

6. purposes according to claim 5, the mass ratio of wherein said pyrroloquinoline quinone disodium salt, thioneine and 5-aminolevulinic acid or its salt is 1:(0.5-15):(0.5-2.0), preferably 1:(0.5-15):(0.75-1.5).

7. Use of pyrroloquinoline quinone disodium salt, ergothioneine and 5-aminolevulinic acid or its salt in the preparation of a product for inhibiting or reducing β-galactosidase.

8. purposes according to claim 7, the mass ratio of wherein said pyrroloquinoline quinone disodium salt, thioneine and 5-aminolevulinic acid or its salt is 1:(0.5-15):(0.5-2.0), preferably 1:(0.5-15):(0.75-1.5).

Images