A composition with anti-aging efficacy based on three-hub long-lasting axle and its application

Abstract

The present application belongs to the technical field of food, and relates to a composition with anti-aging effect based on three-pivot longevity axis and application thereof, wherein the composition comprises ginsenoside, quercetin and ergothioneine. The composition prepared by the present application can target key nodes of the three-pivot longevity axis: quercetin reduces NAD + consumption and protects SIRT1 activity; ergothioneine improves NAD + level and activates SIRT1 through the CSE / H2S / cGPDH pathway; ginsenoside CK promotes NAD + remedial synthesis by activating LKB1 / AMPK to up-regulate NAMPT expression. The three components form a synergistic regulation, activate the AMPK-SIRT1-PGC-1 alpha positive feedback network, and have a synergistic effect on anti-aging effect.

Description

Technical Field

[0001] This invention belongs to the field of food technology and relates to a composition with anti-aging effects based on a three-hub longevity axis and its application, particularly a composition with anti-aging effects based on the synergistic activation of the AMPK-SIRT1-PGC-1α three-hub longevity axis and its application. Background Technology

[0002] Aging is a complex biological process, and the accumulation of senescent cells is a key factor driving aging and age-related diseases. Current research indicates that the combination of dasatinib and quercetin (D+Q) can effectively clear senescent cells; however, dasatinib is a prescription drug with clear safety risks. Among numerous theories of aging mechanisms, nicotinamide adenine dinucleotide (NAD) is considered a potential culprit. + NAD+ levels decline with age and are widely recognized as one of the key molecular events driving the aging process in mammals. + It is a core coenzyme for hundreds of redox reactions in organisms, and also a longevity protein (Sirtuins), a DNA repair enzyme (PARPs), and a multifunctional NAD+. + It is a common substrate for key signaling molecules such as the metabolic enzyme CD38. It plays an irreplaceable role in cellular energy metabolism (glycolysis, tricarboxylic acid cycle, and fatty acid oxidation), DNA damage repair, epigenetic regulation, mitochondrial homeostasis maintenance, and inflammatory response suppression.

[0003] During the aging process, including rodents and humans, NAD+ levels decrease in various tissues (such as the liver, fat, muscle, skin, and brain). + The levels showed a significant downward trend. This NAD... + Progressive depletion of NAD+ is considered a key driver of aging-related phenotypes, including mitochondrial dysfunction, metabolic disorders, accumulated oxidative stress, genomic instability, and stem cell depletion. Therefore, restoring intracellular NAD+ through exogenous intervention is crucial. + Leveling up has become one of the most promising strategies in the current anti-aging field.

[0004] Currently, increasing NAD + The main strategies for reducing NAD+ levels include supplementing with its precursors, such as nicotinamide riboside (NR), nicotinamide mononucleotide (NMN), and nicotinamide (NAM). Among these, NMN and NR act as precursors for NAD+. +Its direct precursor can enter cells via specific transporters and be efficiently converted into NAD. + It has shown potential to improve metabolic function, enhance mitochondrial activity, and delay the progression of various age-related pathologies in multiple animal models. Current NAD... + The supplementation strategy mainly relies on the single pathway of exogenous supplementation of NMN / NR precursors, and cannot address the NAD+ deficiency caused by CD38 upregulation during the aging process. + The problem of accelerated degradation.

[0005] The "AMPK-SIRT1-PGC-1α Longevity Axis" is a core theoretical framework in the field of aging biology in recent years. It refers to a cellular energy sensing and metabolic regulation network composed of three core proteins: AMP-activated protein kinase (AMPK), Sirtuin 1 (SIRT1), and peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α). AMPK acts as an "energy sentinel," sensing changes in the intracellular AMP / ATP ratio, triggering catabolism and inhibiting anabolism; SIRT1 acts as a "NAD" (nervous systemic enzyme activator)... + "Receptor", whose deacetylase activity directly depends on intracellular NAD. + / NADH ratio; PGC-1α acts as the "master switch for mitochondrial biogenesis," converting upstream energy / NAD... + Signals are translated into mitochondrial function. These three elements form a positive feedback network: AMPK enhances NAD+ expression by increasing NAMPT expression. + SIRT1 is indirectly activated at the horizontal level. SIRT1 deacetylates LKB1, promoting its cytoplasmic localization to enhance AMPK phosphorylation. AMPK and SIRT1 synergistically activate PGC-1α through phosphorylation and deacetylation, respectively. PGC-1α activates mitochondrial feedback to increase AMP / ATP and NAD. + Further activate AMPK and SIRT1.

[0006] Studies have shown that NAD+ is involved in aging. + The continued decline is the core driving force behind the overall collapse of the three-hub longevity axis. NAD + The decrease directly weakens SIRT1 activity, leading to excessive acetylation of PGC-1α and loss of transcriptional coactivation function. This results in reduced mitochondrial biogenesis, decreased fatty acid oxidation, and reduced energy metabolism efficiency, preventing AMPK from effectively maintaining NAD through NAMPT. + Synthesis, NAD + Further decline—creating a vicious cycle. More importantly, CD38 expression significantly increases with aging, which is a precursor to NAD+. + The main reason for the decline is aging, and it's not just NAD. + Reduced synthesis further exacerbates NAD. +Consumption spirals out of control. Therefore, relying solely on the NMN / NR pathway to supplement NAD is ineffective. + The precursor cannot block CD38-mediated NAD. + Accelerated depletion also fails to simultaneously repair multiple nodes of the three-hub longevity axis. There is an urgent need to develop a multi-target intervention strategy that can simultaneously target multiple nodes of the three-hub longevity axis and synergistically activate the AMPK-SIRT1-PGC-1α positive feedback network.

[0007] Therefore, it is necessary to develop a method that can efficiently enhance the body's NAD. + Compositions that can level and synergistically regulate multiple aging-related pathways and have good bioavailability are a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0008] In view of the shortcomings of the existing technology, the purpose of this invention is to provide a composition with anti-aging effects based on a three-hub longevity axis and its application, particularly a composition with anti-aging effects based on the synergistic activation of the AMPK-SIRT1-PGC-1α three-hub longevity axis and its application.

[0009] To achieve this objective, the present invention employs the following technical solution:

[0010] In a first aspect, the present invention provides a composition with anti-aging effects based on a three-hub longevity axis, the composition comprising ginsenosides, quercetin and ergothioneine.

[0011] Quercetin is a natural senolytic agent that primarily targets the SIRT1 node. It counteracts the apoptosis resistance acquired by senescent cells by inhibiting anti-apoptotic proteins such as Bcl-2 / Bcl-xL, selectively inducing apoptosis in senescent cells. Simultaneously, as a CD38 inhibitor, quercetin can reduce tissue NAD caused by high CD38 expression in senescent cells. + It degrades; furthermore, its anti-inflammatory activity can inhibit the secretion of the senescence-associated secretory phenotype (SASP), alleviating paracrine senescence effects. Therefore, quercetin works by inhibiting CD38-mediated NAD+ degradation. + Consuming and clearing senescent cells that highly express CD38 protects the SIRT1 co-substrate NAD from its source. + The pool maintains the activity of the core nodes of the three-hub axis. Ergothioneine (EGT) primarily targets NAD in the SIRT1 node. + On the supply side, it enhances NAD through the CSE / H2S / cGPDH pathway. + Level, repairing residual cells' NAD +It decreases and specifically protects the mitochondrial membrane, maintaining mitochondrial function in healthy cells. This pathway is completely independent of the NMN / NR salvage synthesis pathway, providing an alternative NAD for the tri-axis. + The control axis broadens the NAD. + Dimensions of the intervention strategy. Ginsenoside CK primarily targets the AMPK node, restoring the energy-sensing ability of senescent cells by activating the LKB1 / AMPK signaling pathway and maintaining endoplasmic reticulum calcium homeostasis through calmodulin (CALM), thereby protecting the functional integrity of the SIRT1 protein. After AMPK activation, it upregulates NAMPT (NAD...) + Increased expression of NAD+ (rate-limiting enzyme in synthesis) + Levels indirectly activate SIRT1, leading to the formation of AMPK-NAD. + The signal cascade of SIRT1-PGC-1α initiates the positive feedback loop of the three-hub long-life shaft.

[0012] This invention combines quercetin, ergothioneine, and ginsenosides to eliminate senescent cells and repair NAD+. + Synergistic anti-aging effects through leveling and restoring energy sensing. The three ingredients respectively protect SIRT1, NAD... + The three dimensions of supply and AMPK activation simultaneously act on the three-hub longevity axis, reconstructing the AMPK-SIRT1-PGC-1α positive feedback network. Ergothioneine (EGT), as a substrate of cystathionine γ-lyase (CSE), produces hydrogen sulfide (H2S) via an enzymatic reaction. H2S activates cytoplasmic glycerol-3-phosphate dehydrogenase (cGPDH) through persulfidation, enhancing mitochondrial glycerol-3-phosphate dehydrogenase (mGPDH) activity and regulating intracellular NAD. + / NADH ratio, increases NAD + Available levels. This pathway is completely independent of the NMN / NR salvage synthesis pathway. Quercetin, as a natural CD38 inhibitor, competitively inhibits CD38's NAD. + Hydrolytic enzyme activity, reducing NAD38 upregulation during aging. + It accelerates degradation; at the same time, quercetin selectively eliminates senescent cells that highly express CD38, reducing NAD at its source. + Consumption of ginsenoside CK, activated by LKB1 / AMPK, upregulates NAMPT expression and promotes NAD. + The salvage synthesis provides a third auxiliary pathway for the dual-channel system, and the three components form a synergistic regulatory effect, resulting in synergistic enhancement.

[0013] Preferably, the composition comprises, by weight, 1-20 parts of ginsenoside, 10-500 parts of quercetin, and 5-100 parts of ergothioneine.

[0014] Preferably, the composition comprises, by weight, 1-10 parts of ginsenoside, 30-300 parts of quercetin, and 10-80 parts of ergothioneine.

[0015] The mass fractions of ginsenosides can be selected from 1 part, 2 parts, 5 parts, 8 parts, 10 parts, 12 parts, 15 parts, 18 parts, 20 parts, etc.; the mass fractions of quercetin can be selected from 10 parts, 20 parts, 30 parts, 50 parts, 80 parts, 100 parts, 150 parts, 200 parts, 250 parts, 300 parts, 350 parts, 400 parts, 450 parts, 500 parts, etc.; the mass fractions of ergothionein can be selected from 5 parts, 10 parts, 15 parts, 20 parts, 30 parts, 40 parts, 50 parts, 60 parts, 70 parts, 80 parts, 90 parts, 100 parts, etc. Other specific values ​​within the above ranges can be selected, which will not be elaborated here.

[0016] Preferably, the ginsenoside is ginsenoside CK.

[0017] This invention has found that ginsenoside CK has a synergistic effect with quercetin and ergothionein in anti-aging, and that ginsenoside CK is superior to other ginsenosides.

[0018] Preferably, the composition is prepared by a method comprising the following steps: physically mixing the raw materials to obtain the composition.

[0019] In a second aspect, the present invention provides the application of the composition with anti-aging effects based on the three-hub longevity axis according to the first aspect in the preparation of products with anti-aging effects.

[0020] Preferably, the composition with anti-aging effects in the product has a mass percentage content of 10-70%, such as 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, etc. Other specific values ​​within the above range can be selected, and will not be elaborated here.

[0021] Preferably, the dosage form of the product includes tablets, capsules, granules, powders, gummies, oral liquids, or beverages.

[0022] Preferably, the product also includes auxiliary materials.

[0023] Preferably, the excipients include any one or a combination of at least two of the following: solvent, emulsifier, dispersant, wetting agent, binder, stabilizer, colorant, diluent, flavoring agent or filler.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] This invention is the first to propose an all-natural composition for clearing senescent cells based on the AMPK-SIRT1-PGC-1α three-hub longevity axis theoretical framework. The three components target different nodes of the three-hub longevity axis, forming a systematic anti-aging regulatory loop. At the SIRT1 node, quercetin clears senescent cells and inhibits SASP inflammation, while simultaneously reducing NAD by inhibiting CD38. + Accelerated consumption protects SIRT1's co-substrate NAD from the source. + Ergothioneine provides an alternative NAD independent of NMN / NR via the CSE / H2S / cGPDH pathway. + Supply activates SIRT1 deacetylase activity. At the AMPK node, ginsenoside CK activates AMPK via LKB1, rebuilding the cellular energy sensing system. After AMPK activation, NAD+ is increased by upregulating NAMPT. + Horizontally, SIRT1 is indirectly activated, forming an activation cascade from AMPK to SIRT1. At the PGC-1α node, NAD... + After the level is restored, SIRT1 substrate restriction is lifted, and SIRT1 deacetylates multiple lysine sites of PGC-1α, enhancing its transcriptional co-activation activity; AMPK simultaneously phosphorylates Thr-177 and Ser-538 sites of PGC-1α, synergistically activating mitochondrial biogenesis, etc. The three components synergistically enhance NAD from three different dimensions. + At the level, EGT regenerates NAD through the H2S / cGPDH pathway. + Quercetin reduces NAD by inhibiting CD38. + Degradation of ginsenoside CK promotes NAD by upregulating AMPK. + Synthesis. The three components form a complete NAD. + Regulatory closed loop, NAD + Once the level is restored, SIRT1 substrate restriction is lifted, activating the entire anti-aging program. Attached Figure Description

[0026] Figure 1 The test substance affects the NAD5+ levels in zebrafish. + The effect of the horizontal level is shown in the diagram.

[0027] Figure 2 This is a graph showing the effect of the test substance on the relative expression level of Sirt1 in zebrafish.

[0028] Figure 3 This is a graph showing the positive rate of SA-β-Gal. Detailed Implementation

[0029] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention in any way.

[0030] Example 1

[0031] This embodiment provides a composition with anti-aging effects, which includes, by weight parts, 3 parts of ginsenoside CK, 50 parts of quercetin, and 30 parts of ergothioneine.

[0032] Its preparation method includes: physically mixing the raw materials to obtain the product.

[0033] Example 2

[0034] This embodiment provides a composition with anti-aging effects, which includes, by weight parts, 1 part of ginsenoside CK, 20 parts of quercetin and 20 parts of ergothioneine.

[0035] The preparation method is the same as in Example 1.

[0036] Example 3

[0037] This embodiment provides a composition with anti-aging effects, which includes, by weight parts, 6 parts of ginsenoside CK, 200 parts of quercetin, and 60 parts of ergothioneine.

[0038] The preparation method is the same as in Example 1.

[0039] Example 4

[0040] This embodiment provides a composition with anti-aging effects, which differs from Example 1 only in that ginsenoside CK is replaced with ginsenoside Rb1 in an equal amount, while other components and contents remain unchanged.

[0041] The preparation method is the same as in Example 1.

[0042] Example 5

[0043] This embodiment provides a composition with anti-aging effects. The only difference between this composition and Example 1 is that ginsenoside CK is replaced with an equal amount of ginsenoside Rb2, while the other components and contents remain unchanged.

[0044] The preparation method is the same as in Example 1.

[0045] Comparative Example 1

[0046] This comparative example provides a composition with anti-aging effects, which differs from Example 1 only in that it does not contain ginsenoside CK, and its reduced mass is proportionally allocated to the mass of quercetin and ergothioneine.

[0047] The preparation method is the same as in Example 1.

[0048] Comparative Example 2

[0049] This comparative example provides a composition with anti-aging effects, which differs from Example 1 only in that it does not contain quercetin, and the reduced mass of quercetin is proportionally allocated to the mass of ginsenoside CK and ergothioneine.

[0050] The preparation method is the same as in Example 1.

[0051] Comparative Example 3

[0052] This comparative example provides a composition with anti-aging effects, which differs from Example 1 only in that it does not contain ergothioneine, and the reduced mass of ergothioneine is proportionally allocated to the mass of ginsenoside CK and quercetin.

[0053] The preparation method is the same as in Example 1.

[0054] Test Example 1

[0055] Anti-aging effect test

[0056] Zebrafish model

[0057] Modeling method: 3 dpf AB zebrafish, the concentration of D-galactose in the culture medium was 16 mg / mL, D-galactose treatment was carried out for 5 consecutive days, and the culture medium was changed every day;

[0058] Group processing:

[0059] Normal control group: Zebrafish that have not undergone modeling were cultured in ordinary aquaculture water for 8 days;

[0060] Model group: Zebrafish were cultured in ordinary aquaculture water for 8 days after modeling;

[0061] Sample group: Zebrafish after modeling were cultured in aquaculture water containing the sample for 8 days, with a sample concentration of 30 mg / L.

[0062] 1. NAD + Horizontal detection

[0063] After the administration of the drug, the zebrafish were washed, homogenized, and the supernatant was collected. The solution was then processed according to the enhanced NAD3 regimen. + / NADH detection kit (WST-8 method), Beyotime, S0176S operation process.

[0064] NAD + It is a core coenzyme for cellular energy metabolism and participates in multiple anti-aging pathways, including mitochondrial oxidative phosphorylation, DNA repair (PARP), and sirtuins deacetylation. Figure 1In the diagram, * indicates p < 0.05 compared to the model group, ** indicates p < 0.01 compared to the model group, and *** indicates p < 0.001 compared to the model group; # indicates p < 0.05 compared to Example 1, ## indicates p < 0.01 compared to Example 1, and ### indicates p < 0.001 compared to Example 1. The results show that the NAD of the model group... + The levels were significantly lower than those in the control group, indicating successful modeling. After intervention with the composition of this invention, NAD... + The significantly improved levels indicate that the composition of the present invention has excellent anti-aging effects; if ginsenoside CK in Example 1 is replaced, or any component is removed, the effect will be significantly reduced.

[0065] 2. Sirt1 gene expression assay

[0066] RNA was extracted from zebrafish and then reverse transcribed to obtain cDNA.

[0067] Test method:

[0068] qPCR system: 10 μL SYBR Mix, 0.4 μL upstream primer, 0.4 μL downstream primer, 2 μL cDNA template, 7.2 μL enzyme-free water;

[0069] Amplification program: 95℃ pre-denaturation for 3 min; 95℃ for 10 s, 60℃ for 30 s, 40 cycles; melting curve to verify primer specificity; calculation of Sirt1 relative expression level.

[0070] SIRT1 (Sirtuin 1) is NAD + SIRT1, a type of deacetylase, is recognized as a "longevity gene," and its expression is significantly downregulated in aging models. Figure 2 In the figures, * indicates p < 0.05 compared to the model group, ** indicates p < 0.01 compared to the model group, and *** indicates p < 0.001 compared to the model group; # indicates p < 0.05 compared to Example 1, ## indicates p < 0.01 compared to Example 1, and ### indicates p < 0.001 compared to Example 1. The SIRT1 expression level in the model group was significantly lower than that in the control group, indicating successful modeling. After intervention with the composition of this invention, the SIRT1 expression level significantly increased, indicating that the composition of this invention has excellent anti-aging effects; replacing ginsenoside CK in Example 1, or removing any component, would lead to a significant decrease in the effect.

[0071] Test Example 2

[0072] Cell plating and aging modeling

[0073] WI-38 cells were seeded in 6-well culture dishes with a cell confluence of 70%. Except for the normal control group, the remaining cell culture medium was supplemented with bleomycin to a final concentration of 0.15 μg / mL and cultured at 37℃ and 5% CO2 for 4 days. After modeling, the drug-containing culture medium was discarded, and the cells were washed 3 times with sterile PBS to completely remove bleomycin.

[0074] Normal control group: young WI38 cells, in standard complete culture medium;

[0075] Model control group: Senescent cells from the model + conventional complete culture medium;

[0076] Sample group: senescent cells used for modeling + culture medium containing 40 μM composition;

[0077] Incubate at 37℃ for 72 hours without changing the medium.

[0078] Follow the instructions for the Beyotime C0602 SA-β-Gal senescence staining kit. Discard the culture medium, wash the cells once with PBS; add the kit's fixative to each well and fix at room temperature for 15 min.

[0079] Discard the fixative and wash with PBS; prepare SA-β-Gal staining working solution, add 1 mL of staining working solution to each well, seal, and incubate at 37°C in a CO2-free incubator in the dark for 16 h.

[0080] Microscopic images were taken, and five fields of view were randomly selected. The ratio of senescent cells to total cells was counted by blue staining, and the positive rate of SA-β-Gal was calculated.

[0081] SA-β-gal (senescence-associated β-galactosidase) is the most widely accepted biomarker of cellular senescence. Results are as follows... Figure 3 As shown, * indicates p < 0.05 compared to the model group, ** indicates p < 0.01 compared to the model group, and *** indicates p < 0.001 compared to the model group; # indicates p < 0.05 compared to Example 1, ## indicates p < 0.01 compared to Example 1, and ### indicates p < 0.001 compared to Example 1. The experimental results show that the SA-β-Gal positive rate in the model group was significantly higher than that in the control group, indicating successful modeling. After intervention with the composition of the present invention, the SA-β-Gal positive rate significantly decreased, indicating that the composition of the present invention has excellent anti-aging effects; replacing ginsenoside CK in Example 1, or removing any component, would lead to a significant decrease in the effect.

[0082] The above description is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention fall within the protection and disclosure scope of the present invention.

Claims

1. A composition with anti-aging effects based on a three-pivot longevity axis, characterized in that, The composition includes ginsenosides, quercetin, and ergothioneine.

2. The composition with anti-aging effects according to claim 1, characterized in that, The composition comprises, by weight, 1-20 parts of ginsenoside, 10-500 parts of quercetin, and 5-100 parts of ergothioneine.

3. The composition with anti-aging effects according to claim 2, characterized in that, The composition comprises, by weight, 1-10 parts of ginsenoside, 30-300 parts of quercetin, and 10-80 parts of ergothioneine.

4. The composition with anti-aging effects according to any one of claims 1-3, characterized in that, The ginsenoside mentioned is ginsenoside CK.

5. The composition with anti-aging effects according to any one of claims 1-4, characterized in that, The composition is prepared by a method comprising the following steps: physically mixing the raw materials to obtain the composition.

6. The use of the composition with anti-aging effects based on the three-hub longevity axis according to any one of claims 1-5 in the preparation of products with anti-aging effects.

7. The application according to claim 6, characterized in that, The composition with anti-aging effects in the product has a mass percentage of 10-70%.

8. The application according to claim 6 or 7, characterized in that, The dosage forms of the products include tablets, capsules, granules, powders, gummies, oral liquids, or beverages.

9. The application according to any one of claims 6-8, characterized in that, The product also includes auxiliary materials.

10. The application according to claim 9, characterized in that, The excipients include any one or a combination of at least two of the following: solvents, emulsifiers, dispersants, wetting agents, binders, stabilizers, colorants, diluents, flavoring agents, or fillers.

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