Application of ergothioneine or its combinations in the preparation of radiation protection agents

Abstract

This invention relates to the use of ergothioneine or compositions thereof in the preparation of radiation protection agents. One embodiment of the ergothioneine or compositions thereof according to this invention can effectively scavenge radiation-induced DNA damage and reactive oxygen species, protecting normal cells from radiation-induced death. It can be used as a radiation protection agent, effectively preventing or alleviating radiation-related diseases.

Description

Technical Field

[0001] This invention relates to ergothioneine or compositions thereof, and more particularly to the use of ergothioneine or compositions thereof in radiation protection. Background Technology

[0002] Radiation therapy relying on high-energy ionizing radiation (such as X-rays and gamma rays) is one of the three traditional therapies used clinically to treat tumors. More than 60% of cancer patients require radiation therapy as a primary or adjuvant treatment. High-energy ionizing radiation damages biological macromolecules such as DNA, proteins, and lipids by directly ionizing or indirectly generating reactive oxygen species (ROS), thereby killing tumor cells. However, this non-specific killing mechanism inevitably damages healthy tissues surrounding the lesion, causing serious side effects. From the perspective of disease progression, early radiation damage manifests as oxidative stress, cellular senescence, and apoptosis in rapidly turnover cells, subsequently inducing tissue inflammation. Late radiation damage manifests as tissue fibrosis, vascular damage, nerve damage, and organ failure due to excessive repair caused by inflammation.

[0003] Depending on the location of the radiation therapy, radiation-related illnesses can be broadly categorized as follows: 1) Whole-body or large-area (>60%) radiation exposure easily induces myelopathic radiation syndrome; 2) Head and neck radiotherapy can damage the oral mucosa, salivary glands, eyes, and nervous system; 3) Thoracic radiation easily induces radiation damage to organs such as the esophagus, lungs, heart, and liver; 4) Pelvic and abdominal radiotherapy usually leads to radiation damage to the gastrointestinal tract, kidneys, bladder, ovaries, and vagina. These radiation-related illnesses not only severely reduce patients' quality of life but also limit the increase in radiation dose, thus hindering the effectiveness of radiotherapy.

[0004] However, clinical treatment for these radiation-related illnesses remains limited to symptomatic relief. Currently, only one radioprotective agent, amifostine, is approved clinically for relieving xerostomia and stomatitis induced by radiotherapy for head and neck cancer. However, amifostine's extremely short half-life and narrow safety window severely limit its application in high-dose ionizing radiation protection. Therefore, there is an urgent need to develop a broad-spectrum, highly effective, and safe radioprotective agent. Summary of the Invention

[0005] To overcome at least one of the defects of the prior art, in a first aspect, an embodiment of the present invention provides the use of ergothioneine or a composition thereof in the preparation of radiation protection agents.

[0006] According to one embodiment of the present invention, the ergothioneine composition comprises ergothioneine, sodium hyaluronate, and water.

[0007] According to one embodiment of the present invention, the sodium hyaluronate in the ergothioneine composition has a mass percentage content of 0.3% to 2%.

[0008] According to one embodiment of the present invention, the ergothioneine composition is a hydrogel.

[0009] In a second aspect, one embodiment of the present invention provides a radiation protection agent, including ergothioneine or a combination thereof.

[0010] Thirdly, one embodiment of the present invention provides the use of ergothioneine or a composition thereof in the preparation of a medicament for the prevention or treatment of radiation-induced diseases.

[0011] According to one embodiment of the present invention, the ergothioneine composition comprises ergothioneine, sodium hyaluronate, and water; and / or,

[0012] The radiation-related illnesses include radiation-induced gastroenteritis.

[0013] According to one embodiment of the present invention, the radiation-induced disease includes one or more of the following: radiation-induced oral cavity injury, radiation-induced esophageal injury, radiation-induced gastric injury, radiation-induced intestinal injury, radiation-induced lung injury, bone marrow type radiation syndrome, radiation-induced liver injury, radiation-induced kidney injury, radiation-induced bladder injury, radiation-induced ovarian injury, radiation-induced vaginal mucosal injury, radiation-induced eye injury, radiation-induced skin injury, radiation-induced vascular injury, radiation-induced nerve injury, and radiation-induced muscle injury.

[0014] According to one embodiment of the present invention, the radiation disease is damage induced by exposure to ionizing radiation, wherein the radiation source of the ionizing radiation includes one or more of X-ray radiation, gamma-ray radiation, radionuclide radiation, electron radiation, neutron radiation, and proton radiation.

[0015] Fourthly, one embodiment of the present invention provides a medicament for the prevention or treatment of radiation-induced diseases, comprising ergothioneine or a combination thereof.

[0016] Ergothioneine or its composition according to one embodiment of the present invention can effectively eliminate radiation-induced DNA damage and reactive oxygen species, protect normal cells from radiation-induced death, and can be used as a radiation protection agent to effectively prevent or alleviate radiation-induced diseases.

[0017] In this invention, the above-described technical solutions can be combined with each other to achieve more preferred combinations. Other features and advantages of this invention will be set forth in the following description, and some advantages may become apparent from the description or be learned by practicing the invention. The objects and other advantages of this invention can be realized and obtained from what is particularly pointed out in the description and drawings. Attached Figure Description

[0018] The accompanying drawings are for illustrative purposes only and are not intended to limit the scope of the invention.

[0019] in:

[0020] Figure 1 The scavenging rates of 1,1-diphenyl-2-trinitrophenylhydrazine radicals (DPPH) by different concentrations of ergothioneine in Examples 1-1 are shown in the graph.

[0021] Figure 2 The graph shows the scavenging rate of ergothioneine at different concentrations in Examples 1-2 against 2,2-adiazon-bis(3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt radicals (ABTS).

[0022] Figure 3 Examples 1-3 show the effects of different concentrations of ergothioneine on superoxide anion radicals (O2). 2. - The clearance rate graph;

[0023] Figure 4 The graphs show the scavenging effect of different concentrations of ergothioneine on hydroxyl radicals in Examples 1-4.

[0024] Figure 5 The images show the scavenging effect of ergothionein on X-ray-induced intracellular free radicals, as measured by laser scanning confocal microscopy in Examples 1-5.

[0025] Figure 6 The figures show the mitigation of X-ray-induced cellular DNA damage by ergothionein as measured by laser scanning confocal microscopy in Examples 1-6.

[0026] Figure 7 The graph shows the mitigation of X-ray-induced cell viability reduction by ergothionein as measured by the CCK method in Examples 1-7.

[0027] Figure 8 This is a characterization diagram of the intestinal adhesion properties of sodium hyaluronate hydrogel as measured by a small animal in vivo imaging system in Example 2.

[0028] Figure 9 The UV spectrum of the ergothioneine-sodium hyaluronate hydrogel in Example 2 is shown below.

[0029] Figure 10 The infrared spectrum of the ergothioneine-sodium hyaluronate lyophilized gel in Example 2 is shown below.

[0030] Figure 11 The drug release curve of the ergothioneine-sodium hyaluronate hydrogel in Example 2 is shown.

[0031] Figure 12 This is an H&E histopathological staining image of ergothionein-sodium hyaluronate hydrogel intervention for radiation-induced gastroenteritis in Example 2.

[0032] Figure 13 This is an image of Ly6G immunofluorescence staining of ergothioneine-sodium hyaluronate hydrogel intervention for radiation-induced inflammatory infiltration in Example 2.

[0033] Figure 14A , 14B This is a diagram showing the changes in gut microbiota α diversity induced by radiation intervention with ergothioneine-sodium hyaluronate hydrogel in Example 2.

[0034] Figure 15 This is a diagram of the ergothioneine-sodium hyaluronate hydrogel intervention for radiation-induced gut microbiota community disorder in Example 2.

[0035] Figure 16 This is an H&E histopathological staining image of ergothionein-sodium hyaluronate hydrogel intervention for radiation dermatitis in Example 2.

[0036] Figure 17 An optical microscope image of the ergothioneine calcium alginate microspheres of Comparative Example 1.

[0037] Figure 18 The image shows a scanning electron microscope image of the ergothioneine calcium alginate microspheres of Comparative Example 1.

[0038] Figure 19 The standard curve of ergothioneine obtained by ultraviolet spectrophotometer for Comparative Example 1;

[0039] Figure 20 The image shows the H&E histopathological staining of ergothionein calcium alginate microspheres used in Comparative Example 1 to treat radiation enteritis. Detailed Implementation

[0040] The preferred embodiments of the present invention will be described in detail below. The accompanying drawings constitute a part of the present invention and are used together with the embodiments of the present invention to illustrate the principles of the present invention, but are not intended to limit the scope of the present invention.

[0041] One embodiment of the present invention provides the use of ergothioneine (EGT) or a combination thereof in the preparation of radiation protection agents.

[0042] In one embodiment, ergothioneine is a 2-thioimidazolium amino acid, in the levorotatory form, and its chemical formula is as follows:

[0043]

[0044] In one embodiment, ergothioneine can be obtained through biosynthetic fermentation, and its purity can be 99.8%.

[0045] One embodiment of the present invention provides a radiation protection agent, including ergothioneine or a combination thereof.

[0046] One embodiment of the present invention provides the use of ergothioneine or a combination thereof in the preparation of a medicament for the prevention or treatment of radiation-induced diseases.

[0047] In one embodiment, the ergothioneine composition contains 0.01 to 90% by mass, more preferably 0.1 to 70%, and even more preferably 0.5 to 50%, for example 0.02%, 0.05%, 0.08%, 0.1%, 0.2%, 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 60%, and 80%.

[0048] In one embodiment, the ergothioneine composition comprises ergothioneine, sodium hyaluronate, and water. The mass percentage of sodium hyaluronate may be 0.3–2%, more commonly 0.5–1%, for example 0.6%, 0.8%, 1.2%, 1.5%, 1.6%, or 1.8%.

[0049] In one embodiment, the ergothioneine composition may be a hydrogel, and more particularly, a hydrogel comprising ergothioneine and sodium hyaluronate.

[0050] In one embodiment, the ergothioneine composition includes ergothioneine and other types of drugs, including one or more of anti-inflammatory drugs, antibiotics, antiviral drugs, hormonal drugs, immunosuppressants, probiotics, prebiotics, short-chain fatty acids, proteins, peptides, exosomes, and nucleic acid drugs.

[0051] In one embodiment, radiation sickness is damage induced by exposure to ionizing radiation, and the radiation source of ionizing radiation may include one or more of X-ray radiation, gamma-ray radiation, radionuclide radiation, electron radiation, neutron radiation, and proton radiation.

[0052] In one embodiment, radiation-induced diseases include one or more of the following: radiation-induced oral cavity injury, radiation-induced esophageal injury, radiation-induced gastric injury, radiation-induced intestinal injury, radiation-induced lung injury, bone marrow type radiation syndrome, radiation-induced liver injury, radiation-induced kidney injury, radiation-induced bladder injury, radiation-induced ovarian injury, radiation-induced vaginal mucosal injury, radiation-induced eye injury, radiation-induced skin injury, radiation-induced vascular injury, radiation-induced nerve injury, and radiation-induced muscle injury.

[0053] In one embodiment, the dosage form of the drug includes one or more of the following: liquid formulation, semi-solid formulation, solid formulation, gaseous formulation, and micro / nano drug delivery system.

[0054] In one embodiment, the liquid preparation includes one or more of solutions, suspensions, emulsions, injections, eye drops, lotions, and liniments; the injection may be one or more of water-based injections, powder-based injections, and infusions. The emulsion may be one or more of oil-in-water emulsions, water-in-oil emulsions, and double emulsions.

[0055] In one embodiment, the semi-solid preparation includes one or more of gels, ointments, creams, suppositories, and pastes.

[0056] In one embodiment, the solid dosage form includes one or more of capsules, tablets, granules, microgranules, and films. Tablets may be one or more of ordinary tablets, chewable tablets, effervescent tablets, orally disintegrating tablets, dispersible tablets, lozenges, sustained-release tablets, controlled-release tablets, and enteric-coated tablets. Capsules may be one or more of hard capsules, soft capsules, sustained-release capsules, controlled-release capsules, and enteric-coated capsules.

[0057] In one embodiment, the gaseous formulation includes one or more of powder, aerosol, and spray.

[0058] In one embodiment, the propellant of the aerosol includes one or more of chlorofluorocarbons, hydrofluorocarbons, propane, n-butane, isobutane, compressed carbon dioxide, nitrogen, and nitric oxide.

[0059] In one embodiment, the carrier of the powder atomizer includes one or more of lactose, xylitol, mannitol, amino acids, and phospholipids.

[0060] In one embodiment, the micro / nano drug delivery system includes one or more of the following: microspheres, microcapsules, microemulsions, liposomes, nanospheres, nanoparticles, nanocapsules, and nanoemulsions.

[0061] One embodiment of the present invention provides a pharmaceutical composition for the prevention or treatment of radiation-induced diseases, comprising the active ingredient ergothioneine and pharmaceutically acceptable excipients.

[0062] In one embodiment, pharmaceutical excipients for solid dosage forms, categorized by function, may include one or more of the following: fillers, binders, wetting agents, disintegrants, lubricants, flow aids, capsule materials, plasticizers, coating layers, colorants, flavoring agents, retardants, pore-forming agents, and suppository matrices; pharmaceutical excipients for liquid or semi-solid dosage forms, categorized by function, may include one or more of the following: solvents, cosolvents, solubilizers, pH adjusters, osmotic pressure adjusters, suspending agents, dispersants, gelling agents, emulsifiers, oil phase matrices, preservatives, antibacterial agents, and transdermal absorption enhancers; pharmaceutical excipients for gaseous dosage forms, categorized by function, may include one or more of the following: solvents, cosolvents, antioxidants, antibacterial agents, propellants, lubricants, flow aids, antistatic agents, and carriers.

[0063] In one embodiment, the pharmaceutical excipients, classified according to their chemical structure, include one or more of the following: lactose, sucrose, mannitol, sorbitol, calcium sulfate, calcium carbonate, dicalcium phosphate, magnesium oxide, talc, micronized silica gel, starches and their derivatives, celluloses and their derivatives, gum arabic, agar, sodium alginate, guar gum, hyaluronic acid, chitosan, tragacanth gum, pectin, gelatin, albumin, shellac, polyethylene hydrocarbons, polyacrylic acid compounds, polyesters, polyethers, polyamino acids, organosilicon compounds, hydrocarbons, higher fatty acids and their alcohol esters, hydrogenated vegetable oils, polydimethylsiloxane, sodium fatty alcohol sulfates (esters), stearates, oleates, fatty acid sorbitans, polyoxyethylene fatty alcohol ethers, polysorbates, fatty acid glycerides, stearic acid, cocoa butter, phospholipids, and cholesterol.

[0064] In one embodiment, starches and their derivatives include one or more of starch, pregelatinized starch, dextrin, dry starch, sodium carboxymethyl starch, and hydroxyethyl starch. Celluloses and their derivatives include one or more of powdered cellulose, microcrystalline cellulose, sodium carboxymethyl cellulose, croscarmellose sodium carboxymethyl cellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, cellulose acetate, cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, and hydroxypropyl methylcellulose acetate succinate. Polyethylene hydrocarbon compounds include one or more of povidone, croscarmellose, polyvinyl alcohol, polyvinyl alcohol phthalate, ethylene-vinyl acetate copolymer, polyvinyl acetate phthalate, and polyisobutylene pressure-sensitive adhesive. Polyacrylic acid compounds include one or more of carbomer, acrylic resins, sodium polyacrylate, croscarmellose sodium polyacrylate, and polyacrylic acid pressure-sensitive adhesive. Polyester compounds include one or more of polylactic acid, lactic acid-glycolic acid copolymer, polycaprolactone, and dinonyl sebacate. Polyether compounds include one or more of polyethylene glycol, poloxamer, and polyoxyethylene fatty acid esters. Hydrocarbon compounds include one or more of petrolatum, paraffin, and liquid paraffin. Higher fatty acids and their alcohol esters include one or more of lanolin, beeswax, and cetearyl alcohol.

[0065] In one embodiment, the osmotic pressure regulator includes one or more of sodium chloride, mannitol, glucose, phosphate, and acetate.

[0066] In one embodiment, the transdermal absorption enhancer includes one or more of ethanol, propylene glycol, ethyl acetate, dimethyl sulfoxide, dimethylamide, oleic acid, linoleic acid, lauryl alcohol, laurocapram and its homologues, lecithin, urea, salicylic acid, pyrrolidones, menthol, and camphor.

[0067] In one embodiment, the antioxidant includes one or more of vitamin E, butylated hydroxytoluene, ascorbic acid, sulfite, citric acid, and tartaric acid.

[0068] In one embodiment, the preservative includes one or more of methylparaben, ethylparaben, phenol, benzoic acid, sorbic acid, benzalkonium chloride, alkyltrimethylammonium bromide, and chlorobutanol.

[0069] In one embodiment, according to function, the drug includes one or more of the following: ordinary formulation, sustained-release formulation, controlled-release formulation, and targeted formulation.

[0070] In one embodiment, the route of administration of the drug includes one or more of the following: oral administration, oral administration, sublingual administration, inhalation administration, nasal administration, transdermal administration, injection administration, ocular administration, and cavity administration.

[0071] In one embodiment, the dosage of the drug can be 0.001–300 mg / kg body weight, more further 0.1–250 mg / kg, and even more further 5–150 mg / kg. For example, 1 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg, 50 mg / kg, 100 mg / kg, 150 mg / kg, 200 mg / kg, 250 mg / kg, 300 mg / kg, 350 mg / kg, 400 mg / kg, and 450 mg / kg.

[0072] In one embodiment, the drug is administered according to the dosage form, and the timing of drug administration can be before and / or after irradiation. The optimal administration time is from three days before irradiation until a period of time after irradiation. The specific course of treatment depends on the relief of the patient's symptoms.

[0073] In one embodiment, the radiation-induced gastroenteritis model was established by intraperitoneal irradiation of male Balb / C mice (6–8 weeks old). The radiation source included one or more of gamma rays, X-rays, and various particle radiation exposures. The X-ray irradiation dose could be 0.1–50 Gy, more specifically 3–28 Gy, for example 5 Gy, 10 Gy, 15 Gy, 20 Gy, 25 Gy, 30 Gy, 35 Gy, 40 Gy, or 45 Gy.

[0074] In one embodiment, for the prevention or treatment of radiation-induced gastroenteritis, the preferred dosage form of the drug is an oral gel. The oral gel can effectively prolong the retention time of water-soluble small-molecule ergothioneine in the gastrointestinal tract, thereby extending its efficacy. The excipients included in the oral gel may include one or more of thickeners, antioxidants, and pH adjusters. The gel matrix may include one or more of natural polymers, semi-synthetic polymers, and synthetic polymers; preferably, the gel matrix includes one or more of alginate, pectin, gelatin, agar, gum arabic, hyaluronic acid, chitosan and its derivatives, and starch and its derivatives.

[0075] In one embodiment, the radiation dermatitis model was established by localized dorsal irradiation of male Balb / C mice (6–8 weeks old). The radiation source included one or more of gamma rays, X-rays, and various particle beam exposures. The X-ray irradiation dose could be 0.1–90 Gy, more specifically 15–50 Gy, for example 5 Gy, 10 Gy, 20 Gy, 25 Gy, 30 Gy, 35 Gy, 40 Gy, 45 Gy, 55 Gy, 60 Gy, 65 Gy, 70 Gy, 75 Gy, 80 Gy, and 85 Gy.

[0076] The ergothioneine or its composition according to one embodiment of the present invention can effectively alleviate cell and tissue damage caused by radiation and play a therapeutic role in radiation-induced diseases.

[0077] Ergothioneine or its composition according to one embodiment of the present invention can effectively eliminate radiation-induced DNA damage and reactive oxygen species, protect normal cells from radiation-induced death, thereby effectively preventing or alleviating radiation-related diseases, and improving the quality of life and radiation dose threshold of radiotherapy patients.

[0078] The ergothioneine-containing radiation protection agent or drug of one embodiment of the present invention can be administered before radiation occurs to prevent radiation sickness; it can also be administered after radiation occurs to treat radiation sickness; or it can be administered both before and after radiation occurs.

[0079] The ergothioneine-containing radiation protection agent or drug of one embodiment of the present invention is a hydrogel (or oral gel) containing ergothioneine and sodium hyaluronate, which has good protective effects against radiation gastroenteritis and radiation dermatitis.

[0080] The following description, in conjunction with the accompanying drawings and specific embodiments, further illustrates an embodiment of ergothioneine or a composition thereof according to the present invention.

[0081] Example 1-1

[0082] First, a 100 mM DPPH radical solution was prepared using anhydrous ethanol as the solvent. Then, ergothioneine aqueous solutions (EGT solutions) with concentrations of 2 μg / mL, 7 μg / mL, 15 μg / mL, and 30 μg / mL were prepared. Equal volumes of the DPPH radical solution and each concentration of EGT solution were mixed and reacted in the dark for 30 min. The absorbance of each mixture at 517 nm was then measured using a UV spectrophotometer. The DPPH scavenging results of different concentrations of EGT are shown below. Figure 1 As shown. From Figure 1 It can be seen that the scavenging ability of ergothioneine against DPPH free radicals increases in a dose-dependent manner, and even a low concentration of ergothioneine (15 μg / mL) can achieve a scavenging rate of over 75% against DPPH free radicals.

[0083] Examples 1-2

[0084] First, an ABTS radical solution was prepared by reacting 7 mM ABTS aqueous solution and 2.45 mM potassium persulfate at room temperature in the dark for 16 hours. Then, the mixture was diluted 40–50 times with an appropriate volume of PBS to prepare an ABTS radical working solution. Subsequently, ergothioneine aqueous solutions (EGT solutions) with concentrations of 1 μg / mL, 2 μg / mL, 5 μg / mL, and 10 μg / mL were prepared. Equal volumes of the DPPH radical working solution and each concentration of EGT solution were mixed and reacted in the dark for 10 min. The absorbance of each mixture at 734 nm was then measured using a UV spectrophotometer. The scavenging results of different concentrations of EGT on ABTS radicals are shown below. Figure 2 As shown. From Figure 2 It can be seen that the scavenging ability of ergothioneine against ABTS free radicals increases in a dose-dependent manner, and even a low concentration of ergothioneine (5 μg / mL) can achieve a scavenging rate of nearly 100% against ABTS free radicals.

[0085] Examples 1-3

[0086] First, prepare aqueous solutions of 1 mM NADH, 0.25 mM NBT, and 15 μM PMS, and aqueous solutions of 60 μg / mL and 300 μg / mL ergothioneine. Mix 200 μL PBS, 100 μL NADH, 100 μL NBT, and 100 μL ergothioneine, then add 100 μL PMS to initiate O2· - The reaction was initiated. Immediately after the reaction started, the absorbance of each mixture was measured at 560 nm and recorded for 300 s using a UV-vis spectrophotometer. The scavenging capacity of ergothioneine at different working concentrations for superoxide anions was determined as follows: Figure 3 As shown. From Figure 3 It can be seen that the ergothioneine solution with a concentration of 10 μg / mL exhibits a strong scavenging ability against superoxide anion free radicals.

[0087] Examples 1-4

[0088] First, prepare 4 mM FeSO4 solution, 40 mM H2O2 aqueous solution (pH 4, using acetate buffer as solvent), 1 mM TMB solution (DMSO as solvent), and 100 μg / mL and 600 μg / mL ergothioneine aqueous solutions. Then, mix 150 μL TMB, 150 μL H2O2, and 150 μL ergothioneine aqueous solution, add 150 μL FeSO4 solution to initiate the reaction, and react in the dark for 5 min. The absorption curves of each mixed solution in the 550–750 nm range are measured using a UV-Vis spectrophotometer. The scavenging ability of ergothioneine at different working concentrations against hydroxyl radicals is shown in the results below. Figure 4 As shown. From Figure 4 It can be seen that the ergothioneine solution with a concentration of 150 μg / mL exhibits a strong scavenging ability against hydroxyl radicals.

[0089] Examples 1-5

[0090] The scavenging effect of ergothionein on irradiation-induced intracellular reactive oxygen species was investigated using a reactive oxygen species probe (DCFH-DA). Rat small intestinal crypt epithelial cells (IEC-6) were cultured in confocal culture dishes at a density of 2 × 10⁶ cells / dish. 5 After cell adhesion, complete culture medium containing 50 μg / mL ergothioneine was added and incubated for 3 h. Then, appropriate concentrations of DCFH-DA probe and Hoechst 33342 dye were added to pure DMEM, with 1 mL of the above dye solution added to each dish and the cells were treated at 37°C for 30 min. Immediately after 6 Gy irradiation, fluorescence images were acquired using a laser scanning confocal microscope. For detailed results, please refer to [link to relevant documentation]. Figure 5 . Figure 5 In the text, "Ctrl" represents untreated cells, "EGT" represents cells pre-incubated with ergothioneine, "irradiation" represents irradiated cells, "irradiation + EGT" represents cells pre-incubated with ergothioneine + irradiated, and "positive control" represents cells treated with reactive oxygen species inducing agents.

[0091] like Figure 5 As shown, IEC-6 cells irradiated with X-rays produced a large amount of reactive oxygen species, manifesting as a wide-ranging DCF positive signal. In contrast, cells pre-incubated with ergothioneine did not show a significant DCF positive signal, indicating that ergothioneine can effectively scavenge irradiation-induced reactive oxygen species within cells.

[0092] Examples 1-6

[0093] Immunofluorescence assays were used to investigate the mitigation effect of ergothionein on radiation-induced DNA double-strand breaks. IEC-6 cells were cultured in 24-well plates with attached cell spreaders at a density of 5 × 10⁶ cells per well. 4After cell adhesion, complete culture medium containing 50 μg / mL ergothioneine was added and incubated for 3 h, followed by 6 Gy irradiation. After 3 hours of irradiation, cells were fixed with 4% paraformaldehyde, then 200 μL of pore-forming agent (0.2% Triton 100×, 99.8% PBS) was added to create wells for 10 min, followed by blocking with blocking agent (1% Triton 100×, 5% FBS, 94% PBS) for 1 h. Cells were then treated with phosphorylated histone H2AX antibody and incubated overnight at 4°C. After recovering the primary antibody, cells were washed and treated with fluorescent secondary antibody at 37°C for 1 h. Nuclei were stained with Hoechst 33342 dye and then mounted. Finally, fluorescence images were captured using a laser scanning confocal microscope; see [link to results]. Figure 6 . Figure 6 In the text, "Ctrl" represents untreated cells, "EGT" represents cells pre-incubated with ergothioneine, "irradiated" represents irradiated cells, and "irradiated + EGT" represents cells pre-incubated with ergothioneine and then irradiated.

[0094] like Figure 6 As shown, IEC-6 cells irradiated with X-rays exhibited significant DNA damage in their nuclei, manifested by the abundant appearance of γ-H2AX focal points. In contrast, cells pretreated with ergothioneine showed a significant reduction in γ-H2AX positive signals, indicating that ergothioneine can alleviate irradiation-induced DNA damage.

[0095] Examples 1-7

[0096] The effect of ergothionein on alleviating radiation-induced decrease in IEC-6 cell viability was investigated using the CCK assay. IEC-6 cells were cultured at a density of 4 × 10⁶ cells per well. 3 Cells were cultured at high density in 96-well plates. After cell adhesion, they were incubated for 3 hours in complete medium containing 50 μg / mL ergothioneine, followed by irradiation at 0, 2, 6, 8, and 10 Gy. After 40 hours of irradiation, 100 μL of 10% CCK-8 solution was rapidly added to each well, and the cells were incubated for 1 hour. Finally, the absorbance of each well at 450 nm was measured using a microplate reader. See [link to results] for details. Figure 7 . Figure 7 In the text, "Ctrl" indicates irradiated cells, and "EGT" indicates cells that have been irradiated after being pre-incubated with ergothionein.

[0097] like Figure 7 As shown, the cell viability of both the control group (Ctrl) and the pre-irradiation ergothioneine incubation group (EGT) gradually decreased with increasing irradiation dose. However, pre-irradiation ergothioneine incubation significantly improved the viability of IEC-6 cells at the same radiation dose, indicating that ergothioneine exhibits a beneficial radiation protection effect at the cellular level.

[0098] Example 2

[0099] Study on the intestinal adhesion properties of oral sodium hyaluronate (HA) gel

[0100] First, a 0.5 mg / mL aqueous solution of cyanofluorescein (Cy5) was prepared. Second, an appropriate amount of HA was dissolved in a suitable volume of Cy5 solution, allowing it to swell fully to form a 1% HA-Cy5 gel. Then, seven BALB / c mice were randomly divided into three groups, and administered 200 μL of distilled water (one mouse in the control group), Cy5 aqueous solution, and Cy5-HA hydrogel by gavage, respectively. Mice were sacrificed at 2 h, 6 h, and 12 h after gavage, and gastrointestinal tissues and other organs were collected for in vivo fluorescence imaging analysis.

[0101] like Figure 8 As shown, two hours after gavage, Cy5 fluorescence was almost distributed throughout the entire gastrointestinal tract of mice in both the Cy5 aqueous solution group and the Cy5-HA group. However, the gastrointestinal fluorescence signal in the Cy5 aqueous solution group was significantly weaker than that in the Cy5-HA group. This may be because free Cy5 is more easily absorbed into the bloodstream and redistributed to other organs or excreted from the body. Furthermore, two hours after gavage, the strongest fluorescence signal was observed in the stomach of mice in the Cy5-HA group, while the strongest fluorescence signal was observed in the lower jejunum of the Cy5 aqueous solution group, indicating that HA has bioadhesive properties in the gastrointestinal tract.

[0102] Six hours later, the Cy5 aqueous solution group showed almost no Cy5 distribution in the upper digestive tract, with the colon becoming the main accumulation site for Cy5. In contrast, the Cy5-HA group exhibited significant fluorescent signals in the stomach, duodenum, and jejunum, further reflecting the improvement of small molecule retention in the gastrointestinal tract by HA. Furthermore, when the time was extended to 12 hours, the Cy5-HA group showed a significantly stronger fluorescent signal in the gastrointestinal tract compared to the Cy5 aqueous solution group. These results indicate that gastrointestinal adhesive sodium hyaluronate gel has the ability to enhance the retention of water-soluble small molecules in the gastrointestinal tract, and is expected to further enhance its intestinal protective effect by promoting the intestinal retention of ergothioneine.

[0103] Preparation and characterization of ergothionein-sodium hyaluronate hydrogel

[0104] 1. Preparation: Prepare a 5 mg / mL ergothioneine aqueous solution and sonicate it until fully dissolved. Add an appropriate amount of sodium hyaluronate to the sonicated aqueous solution, stir, and let it stand for 4-5 hours to allow it to swell and degas, obtaining the final ergothioneine-sodium hyaluronate hydrogel. The mass percentage of sodium hyaluronate is 1%.

[0105] 2. Ultraviolet Detection: Ergothioneine-sodium hyaluronate hydrogel, ergothioneine aqueous solution, and sodium hyaluronate hydrogel were diluted 500 times with deionized water. The ultraviolet absorption spectra of the three in the 200-300 nm wavelength range were measured and compared. See [link to results] for details. Figure 9 In the diagram, "EGT-HA" represents ergothioneine-sodium hyaluronate hydrogel, "EGT" represents ergothioneine aqueous solution, and "HA" represents sodium hyaluronate hydrogel.

[0106] like Figure 9 As shown, comparing the UV spectra of EGT-HA and EGT solutions of the same concentration reveals that the introduction of HA did not alter the peak position or absorption intensity of EGT's characteristic absorption peak at 257 nm. This indicates that EGT and HA form an EGT-HA gel through physical interaction.

[0107] 3. Infrared Detection: Ergothioneine-sodium hyaluronate lyophilized hydrogel, a powder mixture of ergothioneine and sodium hyaluronate, pure ergothioneine powder, and pure sodium hyaluronate powder were thoroughly ground. An appropriate amount was then taken and thoroughly ground and mixed with potassium bromide powder. The mixture was then compressed into tablets, and the infrared absorption spectra of each sample were measured and compared. For specific results, please refer to [link to relevant documentation]. Figure 10 .

[0108] like Figure 10 As shown, comparing EGT-HA lyophilized gel, a mixture of EGT and HA powder, pure HA powder, and pure EGT powder reveals that the EGT-HA lyophilized gel and the mixture of the two have almost identical infrared spectra. This indicates that the process of HA fully swelling in the EGT solution to form the EGT-HA gel does not involve a chemical reaction, further demonstrating that only a physical interaction exists between the two.

[0109] 4. Drug Release Behavior: First, EGT aqueous solutions with concentrations of 1, 2, 4, 6, 8, 10, and 12 μg / mL were prepared, and the absorbance of each solution at 257 nm was measured to establish a standard curve. Then, three equal volumes of the above EGT-HA gel were placed into dialysis bags with a diameter cutoff of 1000 Da and sealed with clips. The dialysis bags were sequentially immersed in gastrointestinal simulated buffer solutions containing 100 mL of pH 1.2, pH 6.8, and pH 7.4, and released at 100 rpm and 37 °C for 2, 3, and 19 h, respectively. At the selected time points, 2 mL of supernatant was aspirated from the release medium, and 2 mL of the corresponding buffer solution was immediately added. Finally, the absorbance of each supernatant at 257 nm was measured, the cumulative drug release rate at the sampling points was calculated, and the drug release curve of the EGT-HA gel was plotted. See [link to results] for details. Figure 11 .

[0110] The release of EGT from EGT-HA gel while maintaining its inherent chemical structure is a prerequisite for EGT to be absorbed by its specific receptors and exert its radioprotective effect. For example... Figure 11 As shown, EGT-HA gel can cumulatively release approximately 50% of EGT in simulated gastric fluid after two hours, continue to release approximately 30% of EGT in simulated intestinal fluid after three hours, and achieve complete EGT release after approximately 19 hours in simulated colonic fluid. These results indicate that EGT-HA gel can release EGT in all segments of the gastrointestinal tract, providing a basis for its use as a gastrointestinal radiation shield.

[0111] Protective test of ergothioneine-sodium hyaluronate hydrogel against radiation-induced gastroenteritis

[0112] The protective efficacy of the superior formulation of ergothioneine sodium hyaluronate oral gel against radiation-induced gastroenteritis was evaluated using H&E histopathological staining. Six- to eight-week-old male BALB / c rats were randomly assigned to the following groups: 1) Normal group: administered water by gavage, without irradiation; 2) EGT-HA group: administered ergothioneine sodium hyaluronate oral gel by gavage, without irradiation; 3) Irradiation group: administered water by gavage and irradiated the entire abdomen; 4) Irradiation + EGT-HA group: administered ergothioneine sodium hyaluronate oral gel by gavage and irradiated the entire abdomen. Administration was performed by gavage three times every other day before irradiation and every other day after irradiation until sacrifice on the sixth day post-irradiation. One hour before irradiation, rats were administered water or EGT-HA by gavage, followed by anesthesia with 1% sodium pentobarbital solution, and then irradiated into the entire abdominal cavity at parameters of 6 Gy, 160 kV, and 25 mA. Six days after irradiation, mice were sacrificed, and stomach and small intestinal tissues were collected and fixed in paraformaldehyde fixative for 48 hours before being sent for H&E pathological examination. H&E pathological sections were photographed using an inverted fluorescence microscope; for detailed results, please refer to [link to details]. Figure 12 .

[0113] like Figure 12 As shown, on the sixth day after radiation, the gastric pitted epithelium of mice was severely damaged, and the gastric mucosa showed severe erosion. Gastric cells, including mucus neck cells, parietal cells, and chief cells, were severely missing. In contrast, oral administration of ergothioneine sodium hyaluronate gel could greatly alleviate acute radiation gastritis, significantly reducing the degree of epithelial defects and mucosal erosion.

[0114] Compared to the intact and orderly intestinal villi and crypts in the unirradiated group, the intestinal structure of the irradiated mice was severely disrupted by day six, characterized by extensive villus breakage and loss, crypt loss, and inflammatory infiltration. In contrast, oral administration of ergothioneine sodium hyaluronate gel significantly maintained the integrity and height of the villi in irradiated mice and alleviated inflammatory infiltration. Therefore, oral ergothioneine formulations demonstrated a better protective effect against radiation-induced gastroenteritis.

[0115] Ergothioneine-sodium hyaluronate hydrogel alleviates radiation-induced inflammatory infiltration.

[0116] The efficacy of the optimized ergothioneine-sodium hyaluronate hydrogel in alleviating radiation-induced inflammatory infiltration was evaluated using Ly6G immunofluorescence staining. Animal experimental procedures are detailed in the section "Protective Testing of Ergothioneine-Sodium Hyaluronate Hydrogel for Radiation-Induced Gastroenteritis". Mice were sacrificed on the sixth day after irradiation, and small intestinal tissue was fixed in paraformaldehyde fixative for 48 hours and sent to Seville Biotechnology Co., Ltd. for Ly6G immunofluorescence staining. Immunofluorescently stained sections were photographed using a laser scanning confocal microscope; specific results are available in [link to relevant documentation]. Figure 13 .

[0117] like Figure 13 As shown, on day 6 post-irradiation, significant Ly6G signals (white bright spots) appeared in the intestinal mucosa and submucosa, while the Ly6G signal was much weaker in the irradiation + EGT-HA group. Ly6G is a specific marker of neutrophils. Neutrophils are important white blood cells that can promote the progression of radiation enteritis through respiratory bursts. Radiation enteritis can essentially be considered a tissue inflammation characterized by leukocyte infiltration. Therefore, this result indicates that EGT-HA can alleviate the severity of radiation-induced radiation enteritis.

[0118] Ergothioneine-sodium hyaluronate hydrogel alleviates radiation-induced gut microbiota dysbiosis.

[0119] The efficacy of the optimal formulation of ergothioneine-sodium hyaluronate oral gel in alleviating radiation-induced intestinal flora imbalance was evaluated using 16S rDNA sequencing. The animal experimental procedure is detailed in the section "Protective Testing of Ergothioneine-Sodium Hyaluronate Hydrogel for Radiation-Induced Gastroenteritis". Mice were sacrificed on the sixth day after irradiation. Fecal samples from the cecum and colon of each group of mice were collected under aseptic conditions and placed in sterile EP tubes, then rapidly frozen at -80°C until delivery for analysis. Fecal samples from each group were sent to Ouyi Biotechnology Co., Ltd. for 16S rDNA sequencing while preserved on dry ice. Specific results and discussion are as follows.

[0120] Studies have shown that radiation-induced gut microbiota dysbiosis is one of the important mechanisms promoting the development of radiation enteritis. It can exacerbate radiation enteritis by disrupting intestinal barrier permeability, affecting intestinal cell growth and proliferation, and inducing the release of inflammatory factors. Radiation reduces gut microbiota diversity and significantly alters the community structure of gut microbiota. The α-diversity parameter of gut microbiota can be used to assess intraspecific richness and evenness of distribution. Figure 14A , 14BAs shown, on day 6 post-irradiation, the α-diversity parameters of the gut microbiota in the irradiated mice were all low, including the observed species number (left) and Shannon-Wiener index (right), indicating a decrease in gut microbiota diversity after irradiation. Gavage administration of EGT-HA alleviated this decline, suggesting that EGT-HA has the ability to maintain gut microbiota diversity. Furthermore, EGT-HA can positively regulate radiation-induced gut microbiota dysbiosis.

[0121] like Figure 15 As shown, the abundance of Proteobacteria in the gut microbiota of irradiated mice was significantly increased, while the abundance of Bacteroidetes was significantly decreased. Proteobacteria contains many common pathogenic Gram-negative bacteria, such as Escherichia coli, which are important causes of dysbiosis and intestinal inflammation. Conversely, Bacteroidetes, through fermentation of exogenous fiber, produce beneficial short-chain fatty acids, promoting the development and balance of the immune system and exerting anti-inflammatory and anti-infective effects, playing an important role in maintaining intestinal homeostasis. Oral administration of EGT-HA gel increased the abundance of beneficial Bacteroidetes and decreased the abundance of harmful Proteobacteria, indicating that EGT-HA has a positive remodeling ability against irradiation-induced gut microbiota dysbiosis.

[0122] Protective test of ergothioneine-sodium hyaluronate hydrogel against radiation dermatitis

[0123] Six- to eight-week-old male BALB / c mice were divided into four groups for back hair removal: 1) Normal group: no treatment; 2) Irradiation group: the back of the mice was irradiated with an X-ray tube for 90 seconds (50kV, 75μA), and the upper body was sealed with a lead plate; 3) Irradiation + superoxide dismutase (SOD) ointment group; 4) Irradiation + ergothioneine sodium hyaluronate hydrogel (EGT-HA) group. For groups 3) and 4), hair was applied to the back skin of the mice (covering an area of ​​4 cm²) 1 hour before irradiation. 2 Approximately 0.2g of SOD ointment and EGT-HA hydrogel were evenly applied to the mice. The mice were photographed daily for 21 consecutive days. See [link to results] for details. Figure 16 .

[0124] like Figure 16As shown, on day 5 of irradiation, the irradiation group and the irradiation + SOD ointment group showed obvious tissue erythema, while the irradiation + EGT-HA group showed no obvious abnormalities. On day 15 of irradiation, the irradiation-only group and the irradiation + SOD ointment group reached the peak of radiation dermatitis, with large-area moist desquamation and skin bleeding and ulceration. In contrast, the irradiation + EGT-HA group effectively delayed the onset of inflammation, showing only mild moist desquamation. On day 21 of irradiation, the irradiation-only group showed skin ulceration, bleeding, and crusting, while the irradiation + EGT-HA gel and irradiation + SOD ointment groups showed varying degrees of recovery, with the irradiation + EGT-HA gel group showing less damage and faster recovery. This indicates that EGT-HA skin gel can effectively protect against radiation dermatitis.

[0125] Comparative Example 1

[0126] Preparation of calcium alginate microspheres loaded with ergothionein

[0127] Prepare an aqueous solution of 16 mg / mL ergothioneine and sonicate it until fully dissolved. Add an appropriate amount of sodium alginate and pectin powder to the completely dissolved ergothioneine solution and stir at 300 rpm for 1 h to prepare a fully swollen sol containing 4% sodium alginate and 1% pectin (hereinafter referred to as "A4P1@EGT").

[0128] The sol solution was allowed to stand for 1 hour for degassing. Then, the sol solution was transferred to a syringe and microspheres were prepared using a gas shearing method. The peristaltic pump was driven at a speed of 1 mm / min, the gas pressure was 0.1 MPa, and the crosslinking agent was a 400 mM CaCl2 solution. After spraying, the solution was allowed to cure in the crosslinking agent for 20 minutes. Fresh microspheres were collected by centrifugation and gently washed. These fresh microspheres were then lyophilized to obtain A4P1@EGT lyophilized microspheres. The fresh A4P1@EGT microspheres were characterized using an optical microscope, and the lyophilized microspheres were characterized using a scanning electron microscope. The results are shown in [link to results]. Figure 17 , 18 .

[0129] from Figure 17 It can be seen that the A4P1 calcium-based microspheres are relatively uniform in size and regular in shape, with the particle size of fresh microspheres mostly between 5-20 μm. From... Figure 18 It can be seen that after freeze-drying, the A4P1 calcium-based microspheres form relatively regular spherical shapes with a size of about 500 nm.

[0130] Radiation protection efficacy test of calcium alginate microspheres loaded with ergothionein

[0131] EGT aqueous solutions of 1 μg / ml, 3.125 μg / ml, 6.25 μg / ml, 8 μg / ml, 10 μg / ml, and 12.5 μg / ml were prepared, and the absorbance of each solution at the maximum UV absorption wavelength of ergothioneine (256 nm) was measured using a UV spectrophotometer. A standard curve was plotted with concentration on the x-axis and absorbance on the y-axis. The results are shown in [reference needed]. Figure 19 Take the supernatant solution remaining after centrifugation and dilute it appropriately. Measure the absorbance of the diluted supernatant at 256 nm and calculate the encapsulation efficiency of the microspheres. Encapsulation efficiency = (1 - W) / W f / W t )×100%, of which W f W represents the amount of drug free in the supernatant. t This refers to the total amount of drugs used.

[0132] Fresh A4P1@EGT microspheres were prepared according to the above method. The absorbance of the supernatant was measured using a UV spectrophotometer, and the encapsulation efficiency of the microspheres was calculated. Based on the encapsulation efficiency, an appropriate volume of distilled water was added to resuspend the fresh microspheres, bringing the ergothioneine concentration to 5 mg / mL. Six- to eight-week-old BALB / c male rats were randomly divided into four groups: 1) Normal group: administered water by gavage, without irradiation; 2) A4P1@EGT group: administered A4P1@EGT microspheres by gavage, without irradiation; 3) Irradiated group: administered water by gavage and irradiated the entire abdomen; 4) Irradiated + A4P1@EGT group: administered A4P1@EGT by gavage and irradiated the entire abdomen. Administration was performed by gavage three times every other day before irradiation, and by gavage every other day after irradiation until the mice were sacrificed on the sixth day after irradiation. One hour before irradiation, mice were administered water or A4P1@EGT via gavage, followed by anesthesia with 1% sodium pentobarbital solution, and then irradiated into the entire peritoneum at 6 Gy, 160 kV, and 25 mA. Six days post-irradiation, mice were sacrificed, and small intestinal tissue was fixed in paraformaldehyde fixative for 48 hours and sent for H&E pathological examination. H&E pathological sections were photographed using an inverted fluorescence microscope. Detailed results are available in [link to results]. Figure 20 .

[0133] according to Figure 19 As shown, the standard curve for ergothioneine is y = 0.0593x + 0.0684R. 2 =0.9936, and based on this formula, the encapsulation efficiency of A4P1@EGT is approximately 17%. For example... Figure 20 As shown, compared to the intact intestinal villi and orderly arranged crypts in the normal group, the intestinal structure of irradiated mice was severely damaged on day six, characterized by extensive villus breakage and loss, crypt loss, and inflammatory infiltration. In contrast, oral administration of ergothioneine calcium alginate microspheres only showed a relatively general radiation protection effect, manifested by significant shortening and substantial shedding of small intestinal villi.

[0134] Based on the test results above, it can be seen that the ergothioneine-sodium hyaluronate hydrogel of Example 2 has a good protective effect against radiation-induced gastroenteritis and radiation-induced dermatitis. Furthermore, in terms of protection against radiation-induced gastroenteritis, the ergothioneine-sodium hyaluronate hydrogel of Example 2 is more effective than the ergothioneine-loaded calcium alginate microspheres of Comparative Example 1.

[0135] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention.

Claims

1. The use of ergothioneine compositions in the preparation of medicaments for the prevention or treatment of radiation-related diseases; wherein the radiation-related diseases are radiation-induced gastroenteritis and radiation-induced dermatitis; The ergothioneine composition comprises ergothioneine, sodium hyaluronate, and water; The drug is an ergothioneine-sodium hyaluronate hydrogel prepared using an ergothioneine composition, and the preparation method of the ergothioneine-sodium hyaluronate hydrogel includes: First, prepare an aqueous solution of ergothioneine and sonicate it to dissolve it completely. Add an appropriate amount of sodium hyaluronate to the ultrasonically treated aqueous solution, stir, and let stand for 4-5 hours to allow it to fully swell and degas, thus obtaining the final ergothioneine-sodium hyaluronate hydrogel. In the ergothioneine composition, the ergothioneine content is 0.1-5% by mass, and the sodium hyaluronate content is 0.3-2% by mass.

2. The application according to claim 1, wherein, The ergothioneine composition is a hydrogel.

3. The application according to claim 1, wherein, The radiation sickness is damage induced by exposure to ionizing radiation, the radiation source of which includes one or more of X-ray radiation, gamma-ray radiation, radionuclide radiation, electron radiation, neutron radiation, and proton radiation.

4. A radiation protection agent for the prevention or treatment of radiation gastroenteritis and radiation dermatitis, comprising an ergothioneine composition, said ergothioneine composition comprising ergothioneine, sodium hyaluronate and water; The radiation protection agent is an ergothioneine-sodium hyaluronate hydrogel prepared using an ergothioneine composition. The preparation method of the ergothioneine-sodium hyaluronate hydrogel includes: First, prepare an aqueous solution of ergothioneine and sonicate it to dissolve it completely. Add an appropriate amount of sodium hyaluronate to the ultrasonically treated aqueous solution, stir, and let stand for 4-5 hours to allow it to fully swell and degas, thus obtaining the final ergothioneine-sodium hyaluronate hydrogel. In the ergothioneine composition, the ergothioneine content is 0.1-5% by mass, and the sodium hyaluronate content is 0.3-2% by mass.

5. A medicament for the prevention or treatment of radiation sickness, comprising an ergothioneine composition, said ergothioneine composition comprising ergothioneine, sodium hyaluronate and water; The radiation-related diseases mentioned are radiation gastroenteritis and radiation dermatitis; The drug is an ergothioneine-sodium hyaluronate hydrogel prepared using an ergothioneine composition, and the preparation method of the ergothioneine-sodium hyaluronate hydrogel includes: First, prepare an aqueous solution of ergothioneine and sonicate it to dissolve it completely. Add an appropriate amount of sodium hyaluronate to the ultrasonically treated aqueous solution, stir, and let stand for 4-5 hours to allow it to fully swell and degas, thus obtaining the final ergothioneine-sodium hyaluronate hydrogel. In the ergothioneine composition, the ergothioneine content is 0.1-5% by mass, and the sodium hyaluronate content is 0.3-2% by mass.

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