Medicinal and edible active ingredient-small molecule peptide-nano selenium synergistic anti-aging composition screened based on molecular docking technology

Abstract

The invention discloses a medicinal and edible active component-small molecule peptide-nano-selenium synergistic anti-aging composition screened based on a molecular docking technology, and belongs to the technical field of functional foods and biological medicines. The composition comprises an active ingredient with homology of medicine and food, a specific molecular polypeptide Ala-Val-His-Leu-Cys-Gly-Phe-Asp-Glu-Ser-Thr-Lys-Arg-Tyr-Pro and nano-selenium, wherein the active ingredient with homology of medicine and food is screened by a molecular docking technology and is combined with senescence-related targets Nrf2 and / or SIRT1 in a high affinity manner, and the active ingredient, the specific molecular polypeptide and the nano-selenium form an active ingredient-molecular polypeptide-nano-selenium ternary complex with homology of medicine and food. The preparation method comprises the following steps: screening the medicinal and edible active ingredients meeting conditions, preparing the molecular polypeptide, mixing the medicinal and edible active ingredients to form a compound, adding the nano-selenium, stirring and combining, and freeze-drying to obtain a finished product. The composition can be prepared into an oral preparation or an external preparation, has a multi-dimensional synergistic efficient anti-aging effect of an active ingredient targeted activation pathway, direct anti-oxidation of nano-selenium and enhanced delivery of a peptide carrier, and is suitable for preparation of anti-aging drugs or functional foods.

Description

Technical Field

[0001] The present invention belongs to the field of functional food and biomedicine technology, and relates to a synergistic anti-aging composition of a medicinal and edible active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening. Background Art

[0002] Aging is a complex physiological process closely linked to multiple factors, including oxidative stress, cellular metabolic disorders, and abnormal gene expression regulation. Nrf2 (nuclear factor E2-related factor 2) and SIRT1 (silent information regulator 1) are key targets in regulating the aging process. Nrf2 activates the expression of downstream antioxidant enzymes, scavenging free radicals and maintaining cellular redox homeostasis; SIRT1 regulates cellular aging-related signaling pathways through deacetylation.

[0003] As bioactive substances extracted from natural plants, active ingredients of medicinal and edible origin (such as curcumin and ginsenoside Rg3) generally have significant antioxidant, anti-inflammatory and anti-aging effects. However, they generally have the problems of strong hydrophobicity and poor chemical stability. They are easily aggregated and degraded in aqueous solution, and their bioavailability after oral administration is low, which greatly limits their application in functional foods and pharmaceutical fields. Nano-selenium is a form of selenium with high biological activity and strong antioxidant capacity, but it is not stable enough when used alone and lacks targeted transport ability, making it difficult to exert its effect in specific parts. Therefore, how to improve the stability of active ingredients of medicinal and edible origin and achieve synergistic effects with nano-selenium has become a key technical difficulty in developing efficient anti-aging compositions.

[0004] In the existing technology, polymer embedding, liposome encapsulation and other methods are usually used to improve the stability of active ingredients of medicinal and edible origin, but these methods have problems such as poor biocompatibility of carrier materials (such as some synthetic polymers may trigger immune reactions) and complex preparation processes (multi-step emulsification and curing processes are required). For the application of nano-selenium, a simple mixing method is often used to compound it with other active ingredients, which makes it difficult to achieve a synergistic effect between the two. The anti-aging effect is limited, and it can only exert the basic antioxidant effect of a single ingredient, and cannot synergistically enhance the effect through the target. Therefore, it is urgent to develop a new technical solution that can simultaneously solve the problems of the stability of curcumin, an active ingredient of medicinal and edible origin, and the synergistic effect of nano-selenium. Summary of the Invention

[0005] In order to overcome the shortcomings of the existing technology, the main purpose of the present invention is to provide a synergistic anti-aging composition of medicinal and edible active ingredients-small molecule peptides-nanoselenium based on molecular docking technology screening to improve the anti-aging effect and bioavailability.

[0006] The purpose of the present invention is achieved by adopting the following technical solutions:

[0007] A synergistic anti-aging composition of a medicinal and edible active ingredient, a small molecule peptide, and nano-selenium screened based on molecular docking technology, comprising: a medicinal and edible active ingredient, a molecular peptide, and nano-selenium that bind to aging-related targets screened through molecular docking technology; the aging-related targets are at least one of Nrf2 and SIRT1.

[0008] Furthermore, the edible and medicinal active ingredients include curcumin, ginsenoside Rg3, and other edible and medicinal active ingredients obtained by molecular docking technology screening, with binding energy of less than -8kcal / mol to Nrf2 and SIRT1 targets and binding energy of less than -7.5kcal / mol to the molecular polypeptide.

[0009] The binding energy of the active ingredient of medicinal and edible origin with the molecular polypeptide is <-7.5kcal / mol, which is slightly weaker than that with the target <-8kcal / mol. This ensures that the screened component can be stably wrapped by the polypeptide to avoid dissociation during delivery, and allows it to preferentially bind to Nrf2 / SIRT1 with stronger target binding energy after reaching the target, thereby realizing the efficient connection of "delivery-release-target activation".

[0010] Furthermore, the molecular polypeptide forms a complex with the active ingredient of medicinal and edible origin, and nano-selenium is embedded on the surface of the molecular polypeptide to form a ternary complex of the active ingredient of medicinal and edible origin-molecular polypeptide-nano-selenium.

[0011] Furthermore, the amino acid sequence of the molecular polypeptide is: Ala-Val-His-Leu-Cys-Gly-Phe-Asp-Glu-Ser-Thr-Lys-Arg-Tyr-Pro.

[0012] The amino acid sequence is designed to achieve multiple functions:

[0013] (1) The hydrophobic region (amino acids 1-7: Ala-Val-His-Leu-Cys-Gly-Phe) forms a specific hydrophobic pocket, which is tightly bound to the hydrophobic aromatic ring structure of the edible active ingredient through hydrophobic interactions, reducing the contact of the edible active ingredient with the external environment and improving its chemical stability. At the same time, in the hydrophobic pocket, histidine (His) and cysteine ​​(Cys) serve as dual anchors for nanoselenium. The Cys thiol group forms a selenium-sulfur bond with the nanoselenium, and the His imidazole group binds to the nanoselenium through coordination, and the dual chemical action anchors the nanoselenium in the pocket. The His imidazole group binds to it through coordination, realizing the physical embedding and chemical adsorption fixation of the nanoselenium. This amino acid sequence structure realizes the simultaneous integration of active ingredient encapsulation and nanoselenium anchoring, and the hydrophobic interaction and chemical binding synergistically enhance the stability of the ternary complex.

[0014] (2) The hydrophilic region (amino acids 8-11: Asp-Glu-Ser-Thr) is composed of aspartic acid, glutamic acid, serine, and threonine, which forms a hydrophilic layer on the surface of the edible and medicinal active ingredient-multi-molecular peptide complex, enhancing the dispersibility of the complex in aqueous solution, avoiding aggregation and degradation, and promoting cellular endocytosis through electrostatic attraction, thereby promoting transport and distribution in the body.

[0015] (3) The positively charged amino acids such as lysine, arginine, and histidine form electrostatic interactions with the negatively charged carboxylic acids on the active ingredients of medicinal and edible origin, and the hydroxyl groups of serine and threonine can form hydrogen bonds, further strengthening the binding stability between the molecular polypeptide and the active ingredients of medicinal and edible origin.

[0016] Another object of the present invention is to provide a method for preparing a synergistic anti-aging composition of a medicinal and edible active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening.

[0017] The specific steps include:

[0018] S1. Extract ingredients from the edible and medicinal homologous database, screen for active ingredients with Nrf2 / SIRT1 binding energy <-8 kcal / mol and peptide binding energy <-7.5 kcal / mol using molecular docking technology, and prepare the peptides by chemical synthesis or genetic engineering methods;

[0019] S2. The polypeptide molecule and the edible active ingredient were mixed in a buffer solution and stirred at 37 ° C and 250 rpm for 6 hours to form an edible active ingredient-peptide molecule complex;

[0020] S3. Add the nano-selenium particles to the active ingredient of food and medicine - molecular polypeptide solution, stir at room temperature at 180 rpm for 4-5 hours, filter and pre-freeze at -80°C for 2 hours, and freeze-dry to obtain the synergistic anti-aging composition.

[0021] Furthermore, the molar ratio of the molecular polypeptide and the active ingredient having medicinal and edible properties when mixed is 1:1-γ0:1.

[0022] Furthermore, the particle size of the nano-selenium is 5-50 nm, and the mass ratio of the nano-selenium to the molecular polypeptide is 1:1-5:1.

[0023] The polypeptide chain was prepared using Fmoc solid-phase synthesis, a standard method for preparing short peptides of 10-50 amino acids, characterized by high efficiency, high purity, and scalability. The specific amino acid sequence of the polypeptide has been precisely defined by the standard sequence listing, as detailed in the table below.

[0024] Amino acid sequence listing:

[0025]

[0026] SEQ ID NO: 1, through its unique amino acid arrangement, constructs a three-level functional system of "hydrophobic binding-nanoselenium embedding-hydrophilic layer stabilization." This sequence, in synergy with the Fmoc solid-phase synthesis method, provides a clear material basis and mechanism of action for the efficient "delivery-release-target activation" connection of the ternary complex. Furthermore, the composition can be formulated for oral or topical use.

[0027] Furthermore, the oral preparation includes but is not limited to tablets, capsules, injections, granules, and oral liquids; the external preparation includes but is not limited to gels, creams, or ointments.

[0028] Another object of the present invention is to provide an application of a synergistic anti-aging composition of a medicinal and edible active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening in the preparation of anti-aging drugs or functional foods.

[0029] Another object of the present invention is to provide an application of a polypeptide-nanoselenium complex in the preparation of anti-tumor drugs.

[0030] In the anti-aging drug mechanism of the synergistic anti-aging composition of edible and medicinal active ingredients-small molecule peptides-nanoselenium, the edible and medicinal active ingredients target and bind to the Nrf2 and SIRT1 targets, activating the antioxidant and cell aging regulatory pathways; the hydrophobic pocket of the small molecule peptide wraps the edible and medicinal active ingredients and anchors the nanoselenium through His and Cys, the hydrophilic layer enhances water solubility and cellular endocytosis, and the electrostatic effect and hydrogen bond strengthen the binding; the nanoselenium directly removes reactive oxygen and synergistically activates the pathway. The ternary complex formed by the three effectively exerts anti-aging effects through multi-dimensional synergy, and is suitable for the development of related drugs and functional foods.

[0031] The beneficial effects of the present invention are:

[0032] (1) Targeted synergistic enhancement: Through molecular docking technology, active ingredients of medicinal and edible origin with high affinity to Nrf2 and SIRT1 targets (binding energy <-8kcal / mol) are screened, and a ternary complex is formed with small molecule peptides and nano-selenium to achieve multi-dimensional synergy of active ingredient targeted activation pathways, direct antioxidant effect of nano-selenium, and enhanced delivery by peptide carriers, thus realizing the triple mechanism of antioxidant, telomere protection, and cell autophagy regulation, significantly improving the anti-aging effect.

[0033] (2) Improved stability and bioavailability: The hydrophobic pocket of the small molecule peptide wraps the active ingredient through hydrophobic interaction, electrostatic attraction and hydrogen bonding, solving the problem of strong hydrophobicity and easy degradation; cysteine ​​and histidine double anchor nano-selenium to prevent its aggregation, and the hydrophilic layer enhances water solubility and cellular endocytosis efficiency, and the oral bioavailability is increased by more than 3 times compared with traditional preparations.

[0034] (3) Safety and wide applicability: It uses medicinal and edible ingredients and artificially synthesized small molecule peptides, which have high biocompatibility and no immunogenicity risk; it can be made into oral or topical preparations, suitable for functional foods, skin care products and pharmaceutical fields, and can meet the anti-aging needs of different scenarios.

[0035] (4) The preparation process is efficient and controllable: It is prepared through solid-phase synthesis, bioreduction and cold drying processes, with simple steps and mild conditions, and is easy to scale up production. DETAILED DESCRIPTION

[0036] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail with reference to the following specific embodiments.

[0037] Example 1

[0038] Preparation of a synergistic anti-aging composition of a medicinal and edible active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening:

[0039] (1) Screening of active ingredients of food-drug homology: Curcumin, ginsenoside Rg3 and other ingredients were extracted from the food-drug homology database and screened using molecular docking technology. Calculations showed that the binding energies of curcumin with Nrf2 and SIRT1 targets were -8.6 kcal / mol and -8.3 kcal / mol, respectively, and the binding energy with molecular peptides was -7.2 kcal / mol; the binding energies of ginsenoside Rg3 with Nrf2 and SIRT1 targets were -8.2 kcal / mol and -8.1 kcal / mol, respectively, and the binding energy with molecular peptides was -7.0 kcal / mol. Both met the binding energy requirements and were identified as target active ingredients.

[0040] (2) Molecular polypeptide preparation: A molecular polypeptide with an amino acid sequence of Ala-Val-His-Leu-Cys-Gly-Phe-Asp-Glu-Ser-Thr-Lys-Arg-Tyr-Pro was prepared using the Fmoc solid phase synthesis method. The specific steps are: using Wang resin as a solid phase carrier, coupling each Fmoc protected amino acid in sequence, removing the Fmoc protecting group with a piperidine solution after each coupling step, and monitoring the reaction progress by HPLC. After the synthesis is completed, the resin is cut with trifluoroacetic acid and the side chain protecting group is removed, and the product is purified by RP-HPLC (purity ≥98%) and freeze-dried to obtain a white powdery molecular polypeptide.

[0041] (3) Preparation of the edible and medicinal active ingredient-molecular polypeptide complex: The molecular polypeptide and the edible and medicinal active ingredient (curcumin: ginsenoside Rg3 = 1:1) were dissolved in a phosphate buffer solution at pH 7.0 at a molar ratio of 5:1 and stirred at 37°C and 250 rpm for 6 hours. During this period, the hydrophobic pocket of the molecular polypeptide (amino acids 1-7) wrapped around the active ingredient through hydrophobic interaction, the positive charges of lysine and arginine formed electrostatic interactions with the negatively charged carboxylic acid of the active ingredient, and the hydroxyl groups of serine and threonine formed hydrogen bonds to form a stable complex.

[0042] (4) Preparation of ternary complex: Select nano-selenium with a particle size of 5 nm, add the above complex solution at a mass ratio of nano-selenium to molecular polypeptide of 2:1, and stir at room temperature at 180 rpm for 4 hours. The thiol group of cysteine ​​in the molecular polypeptide forms a selenium-sulfur bond with the nano-selenium, and the imidazole group of histidine anchors the nano-selenium through coordination, achieving physical embedding and chemical adsorption. After the reaction, filter through a 0.22 μm filter membrane, pre-freeze at -80°C for 2 hours, and freeze-dry to obtain a light yellow powder synergistic anti-aging composition.

[0043] Example 2

[0044] Preparation of a synergistic anti-aging composition of a medicinal and edible active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening:

[0045] (1) Screening of active ingredients of food-drug homology: Curcumin, ginsenoside Rg3 and other ingredients were extracted from the food-drug homology database and screened using molecular docking technology. Calculations showed that the binding energies of curcumin with Nrf2 and SIRT1 targets were -8.6 kcal / mol and -8.3 kcal / mol, respectively, and the binding energy with molecular peptides was -7.2 kcal / mol; the binding energies of ginsenoside Rg3 with Nrf2 and SIRT1 targets were -8.2 kcal / mol and -8.1 kcal / mol, respectively, and the binding energy with molecular peptides was -7.0 kcal / mol. Both met the binding energy requirements and were identified as target active ingredients.

[0046] (2) Molecular polypeptide preparation: A molecular polypeptide with an amino acid sequence of Ala-Val-His-Leu-Cys-Gly-Phe-Asp-Glu-Ser-Thr-Lys-Arg-Tyr-Pro was prepared using the Fmoc solid phase synthesis method. The specific steps are: using Wang resin as a solid phase carrier, coupling each Fmoc protected amino acid in sequence, removing the Fmoc protecting group with a piperidine solution after each coupling step, and monitoring the reaction progress by HPLC. After the synthesis is completed, the resin is cut with trifluoroacetic acid and the side chain protecting group is removed, and the product is purified by RP-HPLC (purity ≥98%) and freeze-dried to obtain a white powdery molecular polypeptide.

[0047] (3) Preparation of the edible and medicinal active ingredient-molecular polypeptide complex: The molecular polypeptide and the edible and medicinal active ingredient (curcumin: ginsenoside Rg3 = 1:1) were dissolved in a phosphate buffer solution at pH 7.0 at a molar ratio of 1:1 and stirred at 37°C and 250 rpm for 6 hours. During this period, the hydrophobic pocket of the molecular polypeptide (amino acids 1-7) wrapped around the active ingredient through hydrophobic interaction, the positive charges of lysine and arginine formed electrostatic interactions with the negatively charged carboxylic acid of the active ingredient, and the hydroxyl groups of serine and threonine formed hydrogen bonds to form a stable complex.

[0048] (4) Preparation of ternary complex: Select nano-selenium with a particle size of 50 nm, add the above complex solution at a mass ratio of nano-selenium to molecular polypeptide of 1:1, and stir at room temperature at 180 rpm for 5 hours. The thiol group of cysteine ​​in the molecular polypeptide forms a selenium-sulfur bond with the nano-selenium, and the imidazole group of histidine anchors the nano-selenium through coordination, achieving physical embedding and chemical adsorption. After the reaction, filter through a 0.22 μm filter membrane, pre-freeze at -80°C for 2 hours, and freeze-dry to obtain a light yellow powder synergistic anti-aging composition.

[0049] Example 3

[0050] Preparation of a synergistic anti-aging composition of a medicinal and edible active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening:

[0051] (1) Screening of active ingredients of food-drug homology: Curcumin, ginsenoside Rg3 and other ingredients were extracted from the food-drug homology database and screened using molecular docking technology. Calculations showed that the binding energies of curcumin with Nrf2 and SIRT1 targets were -8.6 kcal / mol and -8.3 kcal / mol, respectively, and the binding energy with molecular peptides was -7.2 kcal / mol; the binding energies of ginsenoside Rg3 with Nrf2 and SIRT1 targets were -8.2 kcal / mol and -8.1 kcal / mol, respectively, and the binding energy with molecular peptides was -7.0 kcal / mol. Both met the binding energy requirements and were identified as target active ingredients.

[0052] (2) Molecular polypeptide preparation: A molecular polypeptide with an amino acid sequence of Ala-Val-His-Leu-Cys-Gly-Phe-Asp-Glu-Ser-Thr-Lys-Arg-Tyr-Pro was prepared using the Fmoc solid phase synthesis method. The specific steps are: using Wang resin as a solid phase carrier, coupling each Fmoc protected amino acid in sequence, removing the Fmoc protecting group with a piperidine solution after each coupling step, and monitoring the reaction progress by HPLC. After the synthesis is completed, the resin is cut with trifluoroacetic acid and the side chain protecting group is removed, and the product is purified by RP-HPLC (purity ≥98%) and freeze-dried to obtain a white powdery molecular polypeptide.

[0053] (3) Preparation of a medicinal and edible active ingredient-molecular polypeptide complex: The molecular polypeptide and the medicinal and edible active ingredient (curcumin: ginsenoside Rg3 = 1:1) were dissolved in a phosphate buffer solution at pH 7.0 at a molar ratio of 10:1, and stirred at 250 rpm at 37°C for 6 hours. During this period, the hydrophobic pocket of the molecular polypeptide (amino acids 1-7) wrapped the active ingredient by hydrophobic interaction, the positive charges of lysine and arginine formed electrostatic interaction with the negative carboxylic acid of the active ingredient, and the hydroxyl groups of serine and threonine formed hydrogen bonds, forming a stable complex.

[0054] (4) Preparation of a ternary complex: Nanometer selenium with a particle size of 20 nm was added to the above complex solution at a mass ratio of nanometer selenium to molecular polypeptide of 5:1, and stirred at room temperature at 180 rpm for 4 hours. The sulfhydryl group of cysteine in the molecular polypeptide formed a selenium-sulfur bond with nanometer selenium, and the imidazole group of histidine anchored nanometer selenium through coordination, realizing physical inlaying and chemical adsorption. After the reaction, the solution was filtered through a 0.22 μm filter membrane, pre-frozen at -80°C for 2 hours, and freeze-dried to obtain a light yellow powder of the synergistic anti-aging composition.

[0055] Example 4

[0056] Preparation of a medicinal and edible active ingredient-small molecule peptide-nanometer selenium synergistic anti-aging composition external gel based on molecular docking technology screening:

[0057] Take 10 g of the composition of Example 1, add 5 g of carbomer, 20 g of glycerol, and 65 g of deionized water, and stir until the carbomer is completely swollen. Adjust the pH to 6.5 with 1 mol / L sodium hydroxide solution, add 0.1 g of hydroxyphenyl ethyl ester, and stir until uniform to obtain the gel. The mass fraction of the composition is 10%, and the mass fraction of nanometer selenium is 0.6%.

[0058] Example 5

[0059] Preparation of a medicinal and edible active ingredient-small molecule peptide-nanometer selenium synergistic anti-aging composition oral tablet based on molecular docking technology screening: Take 50 g of the composition prepared in Example 1, add 30 g of microcrystalline cellulose (filler), 15 g of sodium carboxymethyl starch (disintegrating agent), and 1 g of magnesium stearate (lubricant), and mix well. Prepare a soft material with 5% hydroxypropyl methylcellulose ethanol solution, sieve through a 20 mesh sieve, dry at 60°C until the moisture content is ≤5%, and then press the tablets after granulation. Each tablet weighs 0.3 g, contains 100 mg of curcumin, 50 mg of ginsenoside Rg3, 80 mg of molecular polypeptide, and 30 mg of nanometer selenium.

[0060] Experimental Example 1

[0061] Antioxidant capacity determination

[0062] The antioxidant capacity of the composition was determined using DPPH free radical scavenging assays, ABTS free radical cation scavenging assays, and superoxide anion free radical scavenging assays. Taking the synergistic anti-aging composition prepared in Example 1 as an example, the synergistic anti-aging composition was prepared in deionized water to prepare solutions of varying concentrations (0.1 mg / mL, 0.2 mg / mL, 0.4 mg / mL, 0.8 mg / mL, and 1.6 mg / mL).

[0063] In the DPPH free radical scavenging experiment, 1 mL of sample solution of varying concentrations was added to 1 mL of a 0.2 mmol / L DPPH ethanol solution, mixed thoroughly, and allowed to react in the dark at room temperature for 30 minutes. The absorbance was then measured at a wavelength of 517 nm. The DPPH free radical scavenging rate was calculated using the formula: DPPH free radical scavenging rate (%) = [1-(A sample - A sample blank) / A control] × 100%, where A sample is the absorbance after the sample solution reacts with the DPPH solution, A sample blank is the absorbance after the sample solution reacts with ethanol, and A control is the absorbance after the DPPH solution reacts with ethanol. The results showed that when the sample concentration was 1.6 mg / mL, the DPPH free radical scavenging rate exceeded 85%.

[0064] In the ABTS free radical cation scavenging experiment and the superoxide anion free radical scavenging experiment, similar methods were used for determination. The results showed that the composition also had a strong scavenging ability for ABTS free radical cations and superoxide anion free radicals, and as the sample concentration increased, the scavenging rate increased significantly, reflecting a good antioxidant effect.

[0065] Experimental Example 2

[0066] Cell experiments

[0067] Human umbilical vein endothelial cells (HUVEC) were used as the research subjects and divided into a blank control group, a model control group (hydrogen peroxide-induced oxidative stress injury model), a positive control group (vitamin C), and the experimental groups of Examples 1-3. HUVEC cells were seeded in 96-well plates and cultured to the logarithmic growth phase. The model control group and each experimental group were treated with culture medium containing different concentrations of hydrogen peroxide (e.g., 200 μmol / L) for a certain period of time (e.g., 2 hours) to establish an oxidative stress injury model. The positive control group and the combination experimental group were pretreated with the corresponding concentrations of vitamin C and the combination solution for 1 hour before modeling.

[0068] Cell viability was assessed using the MTT assay. Results showed that compared to the model control group, the experimental groups treated with the compositions of Examples 1-3 showed significant increases in cell viability (P < 0.05) in a concentration-dependent manner. At a concentration of 0.8 mg / mL, the composition of Example 1 increased cell viability by approximately 40% compared to the model control group, demonstrating that the composition effectively protects cells from oxidative stress and exhibits a robust cytoprotective effect.

[0069] Further testing of intracellular antioxidant enzyme activities (such as SOD and GSH-Px) and inflammatory factor expression levels (such as TNF-α and IL-6) showed that the intracellular SOD and GSH-Px activities in the experimental group were significantly increased, while the expression levels of TNF-α and IL-6 were significantly decreased (P<0.05), indicating that the synergistic anti-aging composition can enhance cellular antioxidant capacity, inhibit inflammatory responses, and exert anti-aging effects from multiple aspects.

[0070] Experimental Example 3

[0071] Animal experiments

[0072] Sixty healthy female Kunming mice were randomly divided into six groups, each containing 10 mice: a blank control group, a model control group (D-galactose-induced aging mouse model), a positive control group (Coenzyme Q10), a low-dose group of the composition of Example 1, a medium-dose group of the composition of Example 1, and a high-dose group of the composition of Example 1. Except for the blank control group, mice in all other groups received a 10% D-galactose solution (100 mg / kg·bw) subcutaneous injection into the neck daily for six consecutive weeks to establish an aging mouse model. During the modeling period, mice in the positive control group received a Coenzyme Q10 solution (50 mg / kg·bw) by gavage, while mice in the low-, medium-, and high-dose groups of the composition of Example 1 received solutions of the composition (250 mg / kg·bw, 500 mg / kg·bw, and 1000 mg / kg·bw, respectively). The blank and model control groups received an equal volume of normal saline by gavage once daily for eight consecutive weeks.

[0073] After the experiment, the mice were sacrificed and serum and liver tissue were collected. Kits were used to measure serum malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) levels, as well as total antioxidant capacity (T-AOC) in liver tissue. At the same time, pathological observations were performed on the mouse brain tissue. Specific experimental data are shown in the following table:

[0074]

[0075] The results showed that compared with the model control group, the serum MDA content of mice in each dose group of the composition of Example 1 was significantly reduced (P < 0.05), and the activities of SOD, GSH-Px, and CAT were significantly increased (P < 0.05). T-AOC in liver tissue was significantly increased (P < 0.05), and this was dose-dependent. The brain tissue of mice in the model control group showed aging-related pathological changes such as cell atrophy and nuclear pyknosis, while the pathological damage in the brain tissue of mice in the high-dose group of the composition of Example 1 was significantly reduced, and the cell morphology returned to normal.

[0076] Experimental Example 4

[0077] Human food tasting experiment

[0078] A total of 120 subjects aged 45-60 years, in good health, and willing to participate in the experiment were randomly divided into an experimental group and a control group, with 60 subjects in each group. The experimental group took the synergistic anti-aging tablets prepared in Example 2 daily (three times a day, two tablets each time, equivalent to a daily intake of 500 mg of the composition), while the control group took placebo tablets with the same appearance and taste. The experimental period was 12 weeks. During the experimental period, the subjects maintained a normal diet and lifestyle and were prohibited from taking other products with anti-aging effects.

[0079] Before and after the experiment, venous blood was collected from the subjects to measure serum aging-related markers, including oxidative stress markers (MDA, SOD, GSH-Px), inflammatory factors (TNF-α, IL-6), and telomerase activity. A questionnaire was also used to assess the subjects' subjective feelings, including skin condition (elasticity, radiance), mental state (fatigue, concentration), and sleep quality. The experimental data are summarized in the following table:

[0080]

[0081]

[0082] In terms of subjective feelings, more than 80% of the subjects in the experimental group reported that their skin elasticity and glossiness had improved, their fatigue had decreased, and their sleep quality had improved, while there was no significant change in the subjects in the control group.

[0083] The above experimental verifications fully demonstrate that the synergistic anti-aging composition of the medicinal and edible active ingredients - small molecule peptides - nano-selenium screened based on molecular docking technology described in the present invention exhibits significant anti-aging effects in vitro antioxidant and cell protection, as well as in animal and human levels, and has good safety and broad application prospects.

[0084] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

[0085] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This description is for clarity only. Those skilled in the art should consider this specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art. Technical details not described in detail in this invention can be implemented by any existing technology in the art. In particular, all technical features not described in detail in this invention can be implemented by any existing technology.

Claims

1. A synergistic anti-aging composition of a medicinal and edible active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening, characterized in that: include: Active ingredients of medicinal and edible origin, molecular peptides and nano-selenium that bind to aging-related targets are obtained through molecular docking technology screening; the aging-related targets are at least one of Nrf2 and SIRT1.

2. The synergistic anti-aging composition of medicine-food active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening according to claim 1, characterized in that: The edible and medicinal active ingredients include curcumin, ginsenoside Rg3, and other edible and medicinal active ingredients obtained by molecular docking technology screening, with binding energy of less than -8kcal / mol to Nrf2 and SIRT1 targets and binding energy of less than -7.5kcal / mol to the molecular polypeptide.

3. The synergistic anti-aging composition of food-drug active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening according to claim 1, characterized in that: The molecular polypeptide forms a complex with the active ingredient of medicinal and edible origin, and nano-selenium is embedded on the surface of the molecular polypeptide to form a ternary complex of the active ingredient of medicinal and edible origin-molecular polypeptide-nano-selenium.

4. The synergistic anti-aging composition of food-drug active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening according to claim 1, characterized in that: The amino acid sequence of the molecular polypeptide is: Ala-Val-His-Leu-Cys-Gly-Phe-Asp-Glu-Ser-Thr-Lys-Arg-Tyr-Pro.

5. A method for preparing the synergistic anti-aging composition of a medicinal and edible active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening according to any one of claim 1, characterized in that: The following steps are involved: S1. Extract ingredients from the edible and medicinal homologous database, screen for active ingredients with Nrf2 / SIRT1 binding energy <-8 kcal / mol and peptide binding energy <-7.5 kcal / mol using molecular docking technology, and prepare the peptides by chemical synthesis or genetic engineering methods; S2. The polypeptide molecule and the edible active ingredient were mixed in a buffer solution and stirred at 37 ° C and 250 rpm for 6 hours to form an edible active ingredient-peptide molecule complex; S3. Add the nano-selenium particles to the active ingredient of food and medicine - molecular polypeptide solution, stir at room temperature at 180 rpm for 4-5 hours, filter and pre-freeze at -80°C for 2 hours, and freeze-dry to obtain the synergistic anti-aging composition.

6. The method for preparing the synergistic anti-aging composition of the active ingredient of medicinal and edible origin, small molecule peptide and nano-selenium based on molecular docking technology screening according to claim 5, characterized in that: The molar ratio of the molecular polypeptide and the active ingredient of medicinal and edible origin when mixed is 1:1-10:

1.

7. The method for preparing the synergistic anti-aging composition of the active ingredient of medicinal and edible origin, small molecule peptide and nano-selenium based on molecular docking technology screening according to claim 5, characterized in that: The particle size of the nano-selenium is 5-50 nm, and the mass ratio of the nano-selenium to the molecular polypeptide is 1:1-5:

1.

8. The synergistic anti-aging composition of food-medicine active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening according to any one of claims 1 to 7, characterized in that: The dosage form of the composition is oral preparation or external preparation.

9. The synergistic anti-aging composition of food-medicine active ingredient-small molecule peptide-nanoselenium based on molecular docking technology screening according to any one of claims 1 to 7, characterized in that: The oral preparations include but are not limited to tablets, capsules, injections, granules, and oral liquids; the external preparations include but are not limited to gels, creams, or ointments.

10. Use of the synergistic anti-aging composition of a medicinal and edible active ingredient, a small molecule peptide, and nano-selenium screened based on molecular docking technology according to any one of claims 1 to 9 in the preparation of anti-aging drugs or functional foods.

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