Anti-aging capsule containing ergothioneine and preparation method thereof

Abstract

The invention belongs to the field of anti-aging, and particularly relates to an anti-aging capsule containing ergothioneine and a preparation method of the anti-aging capsule. According to the invention, beta-cyclodextrin is used for inclusion of ergothioneine to block an oxidation path; meanwhile, resveratrol-chitosan is used as a slow-release regulation and control layer, burst release is inhibited, and the dual functions of oxidation resistance and light / oxygen blocking on ergothioneine are achieved; finally, an EudragS100 enteric coating is adopted as a gastric acid protection layer, a compact structure is formed when the pH value is smaller than 7, gastric juice permeation is reduced, and therefore a three-level protection system from molecular inclusion to physical isolation and then to targeted slow release is formed.

Description

Technical Field

[0001] The invention relates to the technical field of anti-aging, in particular to an anti-aging capsule containing ergothioneine and a preparation method thereof. Background Art

[0002] Ergothioneine (ERG), a natural antioxidant, has shown significant potential in the anti-aging field. It effectively delays skin and body aging by scavenging free radicals, protecting mitochondrial function, and regulating cellular oxidative stress pathways. However, ERG's inherent chemical instability severely limits its practical application. Specifically, the thioether bond in the ERG molecule is unstable under light, high temperature, and alkaline conditions, and the loss rate exceeds 30% during encapsulation processing.

[0003] These defects not only lead to waste of raw materials and increased costs, but also reduce the anti-aging efficacy of the final product. Therefore, an innovative preparation method is needed to improve the processing stability and bioavailability of ERG to meet the industrialization needs of ergothioneine anti-aging capsules. Summary of the Invention

[0004] In view of the deficiencies in the prior art, the present invention provides an anti-aging capsule containing ergothioneine and a preparation method thereof.

[0005] A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) 1.9-2.1 mol of β-cyclodextrin was added to water, heated to 44-46 ° C and stirred to dissolve to form a 9-11 wt% β-cyclodextrin aqueous solution; 0.9-1.1 mol of ergothioneine was dissolved in anhydrous ethanol to form a 4-5 wt% ergothioneine ethanol solution; under light-proof conditions, the above-mentioned ergothioneine ethanol solution was slowly added dropwise to the β-cyclodextrin aqueous solution at a dropping speed of 0.9-1.1 mL / min, while maintaining the temperature at 44-46 ° C. After stirring for 3-4 hours, the mixture was cooled to 0-4 ° C and allowed to stand for 12-24 hours to allow the inclusion complex to crystallize, filtered, and washed twice with 0-4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake was redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in a 0.9-1wt% acetic acid aqueous solution to prepare a 1.9-2.1wt% chitosan solution; adding resveratrol and propyl gallate to the chitosan solution and stirring to dissolve; continuing to add 0.3-0.8wt% cerium dioxide nanozyme, 180-200W, 60-80KHz ultrasonic dispersion for 4-5min to uniformly disperse it to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight increases by 15-20wt%, thereby obtaining an ergothioneine sustained-release intermediate; (3) dissolving Eudragit S100 resin in an isopropanol / water mixed solvent to prepare a 7.8-8.2 wt% Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the thioneine sustained-release intermediate obtained in step (2) until the weight gain is 25-30%, thereby obtaining thioneine sustained-release granules; (4) The thioneine sustained-release granules obtained in step (3) are filled into capsules to obtain the anti-aging capsules containing thioneine.

[0006] In step (2), the mass ratio of resveratrol to chitosan is 1:(9-10); and the final concentration of propyl gallate is 0.8-1.5 wt%.

[0007] In step (2), the air inlet temperature of the fluidized bed coating machine is 34-36° C., the nozzle diameter is 0.8-1.0 mm, and the spray pressure is 1.5-1.6 bar.

[0008] In step (3), the air inlet temperature of the fluidized bed coating machine is 28-32° C., the nozzle diameter is 0.8-1.0 mm, and the spray pressure is 1.8-2.0 bar.

[0009] In the isopropanol / water mixed solvent of step (3), the volume ratio of isopropanol to water is 70:(28-32).

[0010] In step (3), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:(4-5); the mass ratio of talc to Eudragit S100 resin is 1:(4-5).

[0011] Beneficial effects of the present invention: β-cyclodextrin, with its hydrophobic cavity characteristics, forms a sandwich inclusion structure with ergothioneine at a molar ratio of 1:2, thereby constructing a stable core at the molecular level, completely encapsulating the indole ring of ERG and blocking the oxidation pathway; at the same time, resveratrol-chitosan serves as a sustained-release regulating layer, and the chitosan molecular chain swells in the intestinal fluid to form a hydrogel diffusion barrier, inhibiting sudden release; CeO2 nanozymes neutralize alkaline free radicals (•OH) to maintain the integrity of the isolation layer; resveratrol absorbs ultraviolet light through the conjugated structure, synergistically with propyl gallate By constructing a regeneration cycle, the dual functions of anti-oxidation and light / oxygen barrier of ergothioneine are achieved, and the hydrogen bond-π conjugated complex formed with chitosan densifies the physical barrier network; more importantly, the bioadhesion of chitosan prolongs the intestinal retention time, thereby creating favorable conditions for sustained-release absorption; finally, the Eudragit S100 enteric coating acts as a gastric acid protective layer, forming a dense structure at pH <7, reducing gastric juice penetration, thereby forming a three-level protection system from molecular inclusion to physical isolation to targeted sustained release. DETAILED DESCRIPTION

[0012] Ergothioneine, CAS number: 497-30-3.

[0013] Cerium dioxide nanozyme, product number: YM-NM-025, Changsha Yimo Biotechnology Co., Ltd.

[0014] β-Cyclodextrin, CAS number: 7585-39-9.

[0015] Chitosan, degree of deacetylation 90%, product number: SL20250307H010, Xi'an Shilin Biotechnology Co., Ltd.

[0016] Resveratrol, CAS: 501-36-0.

[0017] Eudragit S100, model: Eudragit S 100, copolymer of methacrylic acid and methyl methacrylate (1:2), Shanghai Dexiang Pharmaceutical Technology Co., Ltd.

[0018] Talc powder, product number: S30618, Shanghai Yuanye Biotechnology Co., Ltd. Example 1

[0019] A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) Add 2 mol of β-cyclodextrin to water, heat to 45 ° C and stir to dissolve to form a 10 wt% β-cyclodextrin aqueous solution; dissolve 1 mol of ergothioneine in anhydrous ethanol to form a 5 wt% ergothioneine ethanol solution; under light-proof conditions, slowly add the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution at a drop rate of 1 mL / min, while maintaining the temperature at 45 ° C. After stirring for 4 hours, cool to 4 ° C and stand for 12 hours to allow the inclusion complex to crystallize, filter, and wash twice with 4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake is redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in a 1 wt % acetic acid aqueous solution to prepare a 2 wt % chitosan solution; adding resveratrol and propyl gallate to the chitosan solution and stirring to dissolve; continuing to add ceria nanozyme, wherein the final concentration of the ceria nanozyme is 0.5 wt %, and ultrasonically dispersing the nanozyme at 200 W and 80 kHz for 5 min to uniformly disperse the nanozyme to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight increases by 20 wt %, thereby obtaining an ergothioneine sustained-release intermediate; (3) dissolving Eudragit S100 resin in an isopropanol / water mixed solvent to prepare an 8 wt% Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly, to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the thioneine sustained-release intermediate obtained in step (2) until the weight gain is 30%, thereby obtaining thioneine sustained-release granules; (4) The thioneine sustained-release granules obtained in step (3) are filled into capsules to obtain the anti-aging capsules containing thioneine.

[0020] The volume ratio of the ergothioneine ethanol solution and the β-cyclodextrin aqueous solution in step (1) is 1:1.

[0021] In step (2), the mass ratio of resveratrol to chitosan is 1:10; and the final concentration of propyl gallate is 1 wt%.

[0022] In step (2), the air inlet temperature of the fluidized bed coating machine is 35° C., the nozzle diameter is 0.8 mm, and the spray pressure is 1.5 bar.

[0023] In step (3), the air inlet temperature of the fluidized bed coating machine is 30° C., the nozzle diameter is 1.0 mm, and the spray pressure is 2.0 bar.

[0024] In the isopropanol / water mixed solvent of step (3), the volume ratio of isopropanol to water is 70:30.

[0025] In step (3), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:5; the mass ratio of talc to Eudragit S100 resin is 1:4. Example 2

[0026] The difference from Example 1 is that the amount of ceria nanozyme added in step (2) is 0.3 wt %; A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) Add 2 mol of β-cyclodextrin to water, heat to 45 ° C and stir to dissolve to form a 10 wt% β-cyclodextrin aqueous solution; dissolve 1 mol of ergothioneine in anhydrous ethanol to form a 5 wt% ergothioneine ethanol solution; under light-proof conditions, slowly add the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution at a drop rate of 1 mL / min, while maintaining the temperature at 45 ° C. After stirring for 4 hours, cool to 4 ° C and stand for 12 hours to allow the inclusion complex to crystallize, filter, and wash twice with 4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake is redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in a 1 wt % acetic acid aqueous solution to prepare a 2 wt % chitosan solution; adding resveratrol and propyl gallate to the chitosan solution and stirring to dissolve; continuing to add ceria nanozyme, wherein the final concentration of the ceria nanozyme is 0.3 wt %, and ultrasonically dispersing the nanozyme at 200 W and 80 kHz for 5 min to uniformly disperse the nanozyme to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight increases by 20 wt %, thereby obtaining an ergothioneine sustained-release intermediate; (3) dissolving Eudragit S100 resin in an isopropanol / water mixed solvent to prepare an 8 wt% Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly, to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the thioneine sustained-release intermediate obtained in step (2) until the weight gain is 30%, thereby obtaining thioneine sustained-release granules; (4) The thioneine sustained-release granules obtained in step (3) are filled into capsules to obtain the anti-aging capsules containing thioneine.

[0027] The volume ratio of the ergothioneine ethanol solution and the β-cyclodextrin aqueous solution in step (1) is 1:1.

[0028] In step (2), the mass ratio of resveratrol to chitosan is 1:10; and the final concentration of propyl gallate is 1 wt%.

[0029] In step (2), the air inlet temperature of the fluidized bed coating machine is 35° C., the nozzle diameter is 0.8 mm, and the spray pressure is 1.5 bar.

[0030] In step (3), the air inlet temperature of the fluidized bed coating machine is 30° C., the nozzle diameter is 1.0 mm, and the spray pressure is 2.0 bar.

[0031] In the isopropanol / water mixed solvent of step (3), the volume ratio of isopropanol to water is 70:30.

[0032] In step (3), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:5; the mass ratio of talc to Eudragit S100 resin is 1:4. Example 3

[0033] The difference from Example 1 is that the amount of ceria nanozyme added in step (2) is 0.8 wt %; A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) Add 2 mol of β-cyclodextrin to water, heat to 45 ° C and stir to dissolve to form a 10 wt% β-cyclodextrin aqueous solution; dissolve 1 mol of ergothioneine in anhydrous ethanol to form a 5 wt% ergothioneine ethanol solution; under light-proof conditions, slowly add the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution at a drop rate of 1 mL / min, while maintaining the temperature at 45 ° C. After stirring for 4 hours, cool to 4 ° C and stand for 12 hours to allow the inclusion complex to crystallize, filter, and wash twice with 4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake is redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in a 1 wt % acetic acid aqueous solution to prepare a 2 wt % chitosan solution; adding resveratrol and propyl gallate to the chitosan solution and stirring to dissolve; continuing to add ceria nanozyme, wherein the final concentration of the ceria nanozyme is 0.8 wt %, and ultrasonically dispersing the nanozyme at 200 W and 80 kHz for 5 min to uniformly disperse the nanozyme to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight increases by 20 wt %, thereby obtaining an ergothioneine sustained-release intermediate; (3) dissolving Eudragit S100 resin in an isopropanol / water mixed solvent to prepare an 8 wt% Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly, to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the thioneine sustained-release intermediate obtained in step (2) until the weight gain is 30%, thereby obtaining thioneine sustained-release granules; (4) The thioneine sustained-release granules obtained in step (3) are filled into capsules to obtain the anti-aging capsules containing thioneine.

[0034] The volume ratio of the ergothioneine ethanol solution and the β-cyclodextrin aqueous solution in step (1) is 1:1.

[0035] In step (2), the mass ratio of resveratrol to chitosan is 1:10; and the final concentration of propyl gallate is 1 wt%.

[0036] In step (2), the air inlet temperature of the fluidized bed coating machine is 35° C., the nozzle diameter is 0.8 mm, and the spray pressure is 1.5 bar.

[0037] In step (3), the air inlet temperature of the fluidized bed coating machine is 30° C., the nozzle diameter is 1.0 mm, and the spray pressure is 2.0 bar.

[0038] In the isopropanol / water mixed solvent of step (3), the volume ratio of isopropanol to water is 70:30.

[0039] In step (3), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:5; the mass ratio of talc to Eudragit S100 resin is 1:4. Example 4

[0040] The difference from Example 1 is that the amount of propyl gallate added in step (2) is 1.5 wt %; A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) Add 2 mol of β-cyclodextrin to water, heat to 45 ° C and stir to dissolve to form a 10 wt% β-cyclodextrin aqueous solution; dissolve 1 mol of ergothioneine in anhydrous ethanol to form a 5 wt% ergothioneine ethanol solution; under light-proof conditions, slowly add the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution at a drop rate of 1 mL / min, while maintaining the temperature at 45 ° C. After stirring for 4 hours, cool to 4 ° C and stand for 12 hours to allow the inclusion complex to crystallize, filter, and wash twice with 4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake is redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in a 1 wt % acetic acid aqueous solution to prepare a 2 wt % chitosan solution; adding resveratrol and propyl gallate to the chitosan solution and stirring to dissolve; continuing to add ceria nanozyme, wherein the final concentration of the ceria nanozyme is 0.5 wt %, and ultrasonically dispersing the nanozyme at 200 W and 80 kHz for 5 min to uniformly disperse the nanozyme to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight increases by 20 wt %, thereby obtaining an ergothioneine sustained-release intermediate; (3) dissolving Eudragit S100 resin in an isopropanol / water mixed solvent to prepare an 8 wt% Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly, to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the thioneine sustained-release intermediate obtained in step (2) until the weight gain is 30%, thereby obtaining thioneine sustained-release granules; (4) The thioneine sustained-release granules obtained in step (3) are filled into capsules to obtain the anti-aging capsules containing thioneine.

[0041] The volume ratio of the ergothioneine ethanol solution and the β-cyclodextrin aqueous solution in step (1) is 1:1.

[0042] In step (2), the mass ratio of resveratrol to chitosan is 1:10; and the final concentration of propyl gallate is 1.5 wt%.

[0043] In step (2), the air inlet temperature of the fluidized bed coating machine is 35° C., the nozzle diameter is 0.8 mm, and the spray pressure is 1.5 bar.

[0044] In step (3), the air inlet temperature of the fluidized bed coating machine is 30° C., the nozzle diameter is 1.0 mm, and the spray pressure is 2.0 bar.

[0045] In the isopropanol / water mixed solvent of step (3), the volume ratio of isopropanol to water is 70:30.

[0046] In step (3), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:5; the mass ratio of talc to Eudragit S100 resin is 1:4. Example 5

[0047] The difference from Example 1 is that the amount of propyl gallate added in step (2) is 0.8 wt %; A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) Add 2 mol of β-cyclodextrin to water, heat to 45 ° C and stir to dissolve to form a 10 wt% β-cyclodextrin aqueous solution; dissolve 1 mol of ergothioneine in anhydrous ethanol to form a 5 wt% ergothioneine ethanol solution; under light-proof conditions, slowly add the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution at a drop rate of 1 mL / min, while maintaining the temperature at 45 ° C. After stirring for 4 hours, cool to 4 ° C and stand for 12 hours to allow the inclusion complex to crystallize, filter, and wash twice with 4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake is redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in a 1 wt % acetic acid aqueous solution to prepare a 2 wt % chitosan solution; adding resveratrol and propyl gallate to the chitosan solution and stirring to dissolve; continuing to add ceria nanozyme, wherein the final concentration of the ceria nanozyme is 0.5 wt %, and ultrasonically dispersing the nanozyme at 200 W and 80 kHz for 5 min to uniformly disperse the nanozyme to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight increases by 20 wt %, thereby obtaining an ergothioneine sustained-release intermediate; (3) dissolving Eudragit S100 resin in an isopropanol / water mixed solvent to prepare an 8 wt% Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly, to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the thioneine sustained-release intermediate obtained in step (2) until the weight gain is 30%, thereby obtaining thioneine sustained-release granules; (4) The thioneine sustained-release granules obtained in step (3) are filled into capsules to obtain the anti-aging capsules containing thioneine.

[0048] The volume ratio of the ergothioneine ethanol solution and the β-cyclodextrin aqueous solution in step (1) is 1:1.

[0049] In step (2), the mass ratio of resveratrol to chitosan is 1:10; and the final concentration of propyl gallate is 0.8 wt%.

[0050] In step (2), the air inlet temperature of the fluidized bed coating machine is 35° C., the nozzle diameter is 0.8 mm, and the spray pressure is 1.5 bar.

[0051] In step (3), the air inlet temperature of the fluidized bed coating machine is 30° C., the nozzle diameter is 1.0 mm, and the spray pressure is 2.0 bar.

[0052] In the isopropanol / water mixed solvent of step (3), the volume ratio of isopropanol to water is 70:30.

[0053] In step (3), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:5; the mass ratio of talc to Eudragit S100 resin is 1:4. Comparative Example 1

[0054] The difference from Example 1 is that: no ceria nanozyme is added in step (2); A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) Add 2 mol of β-cyclodextrin to water, heat to 45 ° C and stir to dissolve to form a 10 wt% β-cyclodextrin aqueous solution; dissolve 1 mol of ergothioneine in anhydrous ethanol to form a 5 wt% ergothioneine ethanol solution; under light-proof conditions, slowly add the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution at a drop rate of 1 mL / min, while maintaining the temperature at 45 ° C. After stirring for 4 hours, cool to 4 ° C and stand for 12 hours to allow the inclusion complex to crystallize, filter, and wash twice with 4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake is redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in a 1 wt % acetic acid aqueous solution to prepare a 2 wt % chitosan solution; adding resveratrol and propyl gallate to the chitosan solution and stirring to dissolve; ultrasonically dispersing at 200 W and 80 kHz for 5 min to uniformly disperse the chitosan to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight increases by 20 wt %, thereby obtaining an ergothioneine sustained-release intermediate; (3) dissolving Eudragit S100 resin in an isopropanol / water mixed solvent to prepare an 8 wt% Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly, to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the thioneine sustained-release intermediate obtained in step (2) until the weight gain is 30%, thereby obtaining thioneine sustained-release granules; (4) The thioneine sustained-release granules obtained in step (3) are filled into capsules to obtain the anti-aging capsules containing thioneine.

[0055] The volume ratio of the ergothioneine ethanol solution and the β-cyclodextrin aqueous solution in step (1) is 1:1.

[0056] In step (2), the mass ratio of resveratrol to chitosan is 1:10; and the final concentration of propyl gallate is 1 wt%.

[0057] In step (2), the air inlet temperature of the fluidized bed coating machine is 35° C., the nozzle diameter is 0.8 mm, and the spray pressure is 1.5 bar.

[0058] In step (3), the air inlet temperature of the fluidized bed coating machine is 30° C., the nozzle diameter is 1.0 mm, and the spray pressure is 2.0 bar.

[0059] In the isopropanol / water mixed solvent of step (3), the volume ratio of isopropanol to water is 70:30.

[0060] In step (3), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:5; the mass ratio of talc to Eudragit S100 resin is 1:4. Comparative Example 2

[0061] The difference from Example 1 is that: propyl gallate is not added in step (2); A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) Add 2 mol of β-cyclodextrin to water, heat to 45 ° C and stir to dissolve to form a 10 wt% β-cyclodextrin aqueous solution; dissolve 1 mol of ergothioneine in anhydrous ethanol to form a 5 wt% ergothioneine ethanol solution; under light-proof conditions, slowly add the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution at a drop rate of 1 mL / min, while maintaining the temperature at 45 ° C. After stirring for 4 hours, cool to 4 ° C and stand for 12 hours to allow the inclusion complex to crystallize, filter, and wash twice with 4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake is redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in a 1 wt % acetic acid aqueous solution to prepare a 2 wt % chitosan solution; adding resveratrol to the chitosan solution and stirring to dissolve; continuing to add ceria nanozyme, wherein the final concentration of the ceria nanozyme is 0.5 wt %, and ultrasonically dispersing the nanozyme at 200 W and 80 kHz for 5 min to uniformly disperse the nanozyme to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight increases by 20 wt %, thereby obtaining an ergothioneine sustained-release intermediate; (3) dissolving Eudragit S100 resin in an isopropanol / water mixed solvent to prepare an 8 wt% Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly, to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the thioneine sustained-release intermediate obtained in step (2) until the weight gain is 30%, thereby obtaining thioneine sustained-release granules; (4) The thioneine sustained-release granules obtained in step (3) are filled into capsules to obtain the anti-aging capsules containing thioneine.

[0062] The volume ratio of the ergothioneine ethanol solution and the β-cyclodextrin aqueous solution in step (1) is 1:1.

[0063] The mass ratio of resveratrol to chitosan in step (2) is 1:10.

[0064] In step (2), the air inlet temperature of the fluidized bed coating machine is 35° C., the nozzle diameter is 0.8 mm, and the spray pressure is 1.5 bar.

[0065] In step (3), the air inlet temperature of the fluidized bed coating machine is 30° C., the nozzle diameter is 1.0 mm, and the spray pressure is 2.0 bar.

[0066] In the isopropanol / water mixed solvent of step (3), the volume ratio of isopropanol to water is 70:30.

[0067] In step (3), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:5; the mass ratio of talc to Eudragit S100 resin is 1:4. Comparative Example 3

[0068] The difference from Example 1 is that: there is no resveratrol-chitosan layer; A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) Add 2 mol of β-cyclodextrin to water, heat to 45 ° C and stir to dissolve to form a 10 wt% β-cyclodextrin aqueous solution; dissolve 1 mol of ergothioneine in anhydrous ethanol to form a 5 wt% ergothioneine ethanol solution; under light-proof conditions, slowly add the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution at a drop rate of 1 mL / min, while maintaining the temperature at 45 ° C. After stirring for 4 hours, cool to 4 ° C and stand for 12 hours to allow the inclusion complex to crystallize, filter, and wash twice with 4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake is redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving Eudragit S100 resin in an isopropanol / water mixed solvent to prepare an 8 wt% Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly, to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight gain is 30%, thereby obtaining ergothioneine sustained-release granules; (3) The thioneine sustained-release granules obtained in step (2) are filled into capsules to obtain the anti-aging capsules containing thioneine.

[0069] The volume ratio of the ergothioneine ethanol solution and the β-cyclodextrin aqueous solution in step (1) is 1:1.

[0070] In step (2), the air inlet temperature of the fluidized bed coating machine is 30° C., the nozzle diameter is 1.0 mm, and the spray pressure is 2.0 bar.

[0071] In the isopropanol / water mixed solvent of step (2), the volume ratio of isopropanol to water is 70:30.

[0072] In step (2), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:5; the mass ratio of talc to Eudragit S100 resin is 1:4. Comparative Example 4

[0073] The difference from Example 1 is: no Eudragit S100 enteric coating; A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) Add 2 mol of β-cyclodextrin to water, heat to 45 ° C and stir to dissolve to form a 10 wt% β-cyclodextrin aqueous solution; dissolve 1 mol of ergothioneine in anhydrous ethanol to form a 5 wt% ergothioneine ethanol solution; under light-proof conditions, slowly add the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution at a drop rate of 1 mL / min, while maintaining the temperature at 45 ° C. After stirring for 4 hours, cool to 4 ° C and stand for 12 hours to allow the inclusion complex to crystallize, filter, and wash twice with 4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake is redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in a 1 wt % acetic acid aqueous solution to prepare a 2 wt % chitosan solution; adding resveratrol and propyl gallate to the chitosan solution and stirring to dissolve; continuing to add ceria nanozyme, wherein the final concentration of the ceria nanozyme is 0.5 wt %, and ultrasonically dispersing the nanozyme at 200 W and 80 kHz for 5 min to uniformly disperse the nanozyme to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight increases by 20 wt %, thereby obtaining an ergothioneine sustained-release intermediate; (3) The thioneine sustained-release intermediate obtained in step (2) is filled into capsules to obtain the anti-aging capsules containing thioneine.

[0074] The volume ratio of the ergothioneine ethanol solution and the β-cyclodextrin aqueous solution in step (1) is 1:1.

[0075] In step (2), the mass ratio of resveratrol to chitosan is 1:10; and the final concentration of propyl gallate is 1 wt%.

[0076] In step (2), the air inlet temperature of the fluidized bed coating machine is 35° C., the nozzle diameter is 0.8 mm, and the spray pressure is 1.5 bar. Comparative Example 5

[0077] The difference from Example 1 is that resveratrol is replaced by vitamin E; A preparation method for an anti-aging capsule containing ergothioneine comprises the following steps: (1) Add 2 mol of β-cyclodextrin to water, heat to 45 ° C and stir to dissolve to form a 10 wt% β-cyclodextrin aqueous solution; dissolve 1 mol of ergothioneine in anhydrous ethanol to form a 5 wt% ergothioneine ethanol solution; under light-proof conditions, slowly add the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution at a drop rate of 1 mL / min, while maintaining the temperature at 45 ° C. After stirring for 4 hours, cool to 4 ° C and stand for 12 hours to allow the inclusion complex to crystallize, filter, and wash twice with 4 ° C anhydrous ethanol to remove unincluded ERG to obtain a filter cake; finally, the filter cake is redispersed in water and freeze-dried to obtain a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in a 1 wt % acetic acid aqueous solution to prepare a 2 wt % chitosan solution; adding vitamin E and propyl gallate to the chitosan solution and stirring to dissolve; continuing to add ceria nanozyme, wherein the final concentration of the ceria nanozyme is 0.5 wt %, and ultrasonically dispersing the nanozyme at 200 W and 80 kHz for 5 min to uniformly disperse the nanozyme to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) until the weight increases by 20 wt %, thereby obtaining an ergothioneine sustained-release intermediate; (3) dissolving Eudragit S100 resin in an isopropanol / water mixed solvent to prepare an 8 wt% Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly, to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the thioneine sustained-release intermediate obtained in step (2) until the weight gain is 30%, thereby obtaining thioneine sustained-release granules; (4) The thioneine sustained-release granules obtained in step (3) are filled into capsules to obtain the anti-aging capsules containing thioneine.

[0078] The volume ratio of the ergothioneine ethanol solution and the β-cyclodextrin aqueous solution in step (1) is 1:1.

[0079] In step (2), the mass ratio of vitamin E to chitosan is 1:10; and the final concentration of propyl gallate is 1 wt%.

[0080] In step (2), the air inlet temperature of the fluidized bed coating machine is 35° C., the nozzle diameter is 0.8 mm, and the spray pressure is 1.5 bar.

[0081] In the isopropanol / water mixed solvent of step (3), the volume ratio of isopropanol to water is 70:30.

[0082] In step (3), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:5; the mass ratio of talc to Eudragit S100 resin is 1:4. Test Example 1

[0083] High temperature stability test: The anti-aging capsules containing thioneine prepared in the examples and comparative examples were placed under 40°C / 75%RH conditions for 6 months, and the thioneine residual rate was determined by HPLC. The results are shown in Table 1. Table 1: High temperature stability test results Test Example 2

[0084] Light stability test: The anti-aging capsules containing thioneine obtained in Example and Comparative Example were placed in 4500 lux illumination for 48 h, and the thioneine residual rate was measured by HPLC. The results are shown in Table 2; Table 1: Light stability test results Test Example 3

[0085] In vitro simulated gastric fluid and intestinal fluid release behavior test: Simulated gastric fluid: 0.1 mol / L HCl solution + 0.2 wt% NaCl; Simulated intestinal fluid: 0.05 mol / L phosphate buffer; One anti-aging capsule containing ergothioneine prepared in the embodiment and the comparative example was added to 900 mL of simulated gastric fluid at 37° C., stirred at 50 rpm, and the timing was started (t=0); At 120 min, 5 mL of the sample was automatically sampled, filtered through a 0.22 μm filter membrane, and the reaction was immediately terminated in an ice bath; At the end of 2 h, all gastric fluid medium was automatically drained (residual volume < 1%), and 900 mL of simulated intestinal fluid at 37 °C was immediately injected. The switching time was controlled within 2 min. 5 mL of the intestinal fluid phase was sampled 5 min later, and an equal volume of fresh simulated intestinal fluid was added at the same temperature. The sample was filtered through a 0.22 μm filter membrane, and the reaction was immediately terminated in an ice bath. HPLC detection: The mobile phase was acetonitrile-0.1% phosphoric acid aqueous solution in a volume ratio of 20:80, and the detection wavelength was 254 nm; Gastric protection assessment: Calculate the 2h cumulative release rate (Rg): (%) = (total amount of ERG released in gastric juice phase / labeled amount of ERG in capsule) × 100%; Evaluation of sustained-release properties in intestinal fluid: Burst Index: = (5-minute release amount in intestinal fluid phase / total release amount) × 100%; The results are shown in Table 3; Table 3: Gastric and intestinal dissolution test results

[0086] As can be seen from Table 1, Table 2, Table 3, the anti-aging capsule containing thioneine prepared in the embodiment of the present invention, its high temperature, light stability and stomach and intestinal fluid sustained-release performance are significantly better than comparative example. The present invention believes that this is due to the present invention using triple dynamic protection system to improve photothermal stability and the sustained-release regulation of thioneine.

[0087] First, the Eudragit S100 enteric coating acts as a gastric acid protective layer, forming a dense structure at pH <7, thereby reducing gastric juice penetration. The Rg of Example 1 is 4.2%, while that of Comparative Example 4 is 98.7%.

[0088] Secondly, resveratrol-chitosan serves as a sustained-release regulating layer, and the chitosan molecular chains swell in the intestinal fluid to form a hydrogel diffusion barrier, inhibiting burst release; CeO2 nanozymes neutralize alkaline free radicals (•OH) to maintain the integrity of the isolation layer; resveratrol absorbs ultraviolet light through a conjugated structure and cooperates with propyl gallate to construct a regeneration cycle, achieving the dual functions of antioxidant and light / oxygen barrier for ergothioneine. The BI (5 min burst release) of Example 1 is only 8.1, while the BI (5 min burst release) of Comparative Example 3, which does not have a resveratrol-chitosan layer, is 61.8.

[0089] Finally, β-cyclodextrin locks the ergothioneine molecules through hydrophobic interaction, delaying oxidation.

[0090] Comparison between Comparative Example 5 and the Examples also shows that when vitamin E is used instead of resveratrol, its high temperature and light stability and gastric and intestinal fluid sustained release performance are significantly lower than those of Example 1, indicating that a single antioxidant cannot replace the multifunctional effects of resveratrol. Comparative Example 2 lacks propyl gallate, and its performance is also significantly weaker than that of Example 1, indicating that resveratrol and propyl gallate have a synergistic effect, and it is difficult for a single component to achieve its function.

[0091] In summary, the present invention improves the photothermal stability and sustained-release regulation of ergothioneine through a triple dynamic protection system, and realizes the full protection of ERG during oral delivery for the first time.

Claims

1. A preparation method for an anti-aging capsule containing ergothioneine, characterized in that, The following steps are involved: (1) adding β-cyclodextrin to water, heating and stirring to dissolve, to form a β-cyclodextrin aqueous solution; dissolving ergothioneine in anhydrous ethanol to form an ergothioneine ethanol solution; slowly adding the above ergothioneine ethanol solution dropwise to the β-cyclodextrin aqueous solution, stirring at 44-46°C, reacting, cooling and standing, filtering, washing, and obtaining a filter cake; finally, redispersing the filter cake in water, freeze-drying, and obtaining a β-cyclodextrin-ergothioneine inclusion complex; (2) dissolving chitosan in an acetic acid aqueous solution to prepare a chitosan solution; adding resveratrol and propyl gallate to the chitosan solution and stirring to dissolve; continuing to add cerium dioxide nanozyme and ultrasonically dispersing it uniformly to obtain a resveratrol-chitosan isolation layer; finally, using a fluidized bed coating machine, spraying the resveratrol-chitosan isolation layer uniformly onto the β-cyclodextrin-ergothioneine inclusion complex obtained in step (1) to obtain an ergothioneine sustained-release intermediate; (3) dissolving Eudragit S100 resin in an isopropyl alcohol / water mixed solvent to prepare a Eudragit S100 resin solution; adding triethyl citrate and talc, stirring evenly to obtain a Eudragit S100 enteric coating; finally, using a fluidized bed coating machine, spraying the Eudragit S100 enteric coating evenly onto the thioneine sustained-release intermediate obtained in step (2) to obtain thioneine sustained-release granules; (4) The thioneine sustained-release granules obtained in step (3) are filled into capsules to obtain the anti-aging capsules containing thioneine.

2. the preparation method of the anti-aging capsule containing thioneine as claimed in claim 1, is characterized in that, The dropping speed in step (1) is 0.9-1.1 mL / min.

3. the preparation method of the anti-aging capsule containing thioneine as claimed in claim 1, is characterized in that, The power of the ultrasound in step (2) is 180-200W and the frequency is 60-80KHz.

4. the preparation method of the anti-aging capsule containing thioneine as claimed in claim 1, is characterized in that, In step (2), the mass ratio of resveratrol to chitosan is 1:(9-10); and the final concentration of propyl gallate is 0.8-1.5 wt%.

5. the preparation method of the anti-aging capsule containing thioneine as claimed in claim 1, is characterized in that, In step (2), the air inlet temperature of the fluidized bed coating machine is 34-36° C., the nozzle diameter is 0.8-1.0 mm, and the spray pressure is 1.5-1.6 bar.

6. the preparation method of the anti-aging capsule containing thioneine as claimed in claim 1, is characterized in that, In step (3), the air inlet temperature of the fluidized bed coating machine is 28-32° C., the nozzle diameter is 0.8-1.0 mm, and the spray pressure is 1.8-2.0 bar.

7. the preparation method of the anti-aging capsule containing thioneine as claimed in claim 1, is characterized in that, In the isopropanol / water mixed solvent of step (3), the volume ratio of isopropanol to water is 70:(28-32).

8. the preparation method of the anti-aging capsule containing thioneine as claimed in claim 1, is characterized in that, In step (3), the mass ratio of triethyl citrate to Eudragit S100 resin is 1:(4-5); the mass ratio of talc to Eudragit S100 resin is 1:(4-5).

9. An anti-aging capsule containing ergothioneine, characterized in that, The anti-aging capsule containing ergothioneine is prepared by the preparation method of any one of claims 1 to 8.

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