Application of chlorogenic acid and blue light in the preparation of anti-photoaging products and thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid

Abstract

The present invention belongs to the field of biomedicine technology, and discloses the application of chlorogenic acid and blue light in the preparation of anti-photoaging products, and the application of chlorogenic acid-loaded thermosensitive transdermal delivery hydrogel based on the application, and the application in the preparation of anti-skin photoaging drugs, auxiliary anti-skin photoaging health products, tissue repair or regeneration drugs / materials, as well as a method for inhibiting photoaging, which mainly achieves the inhibition of photoaging based on the mitochondrial photobiological effect of chlorogenic acid. The present invention provides the application of chlorogenic acid and blue light in combination, including the application in the preparation of anti-photoaging products; and the application in the preparation of products that promote the generation of adenosine triphosphate. The thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid provided by the present invention releases chlorogenic acid through long-term thermosensitive transdermal release, combined with the mitochondrial photobiological effect, to promote mitochondrial ATP generation, promote the repair of photoaged cells, achieve the effect of promoting the repair of photoaged skin, and achieve the purpose of treating skin photoaging.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and particularly relates to the application of chlorogenic acid and blue light in the preparation of anti-photoaging products, and a thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid based on the application, and its application in the preparation of anti-skin photoaging drugs, auxiliary anti-skin photoaging health products, tissue repair or regeneration drugs / materials, as well as a method for inhibiting photoaging, which mainly achieves inhibition of photoaging based on the mitochondrial photobiological effect of chlorogenic acid. Background Art

[0002] Population aging is a current challenge facing China, and skin aging is the most important factor in determining how individuals assess their aging. According to statistics, over 80% of photoaging is caused by ultraviolet rays. Skin photoaging can cause wrinkles, hyperpigmentation, increased reactive oxygen species (ROS), and skin sagging, increasing the incidence of aging-related diseases. This can be attributed to factors such as DNA damage, oxidative stress, inflammation, decreased collagen synthesis, and mitochondrial dysfunction. Currently, the mechanisms of skin photoaging are not fully understood.

[0003] The main methods currently used to treat skin photoaging, such as laser therapy, chemical peels, and injectable fillers, have limitations such as significant side effects, long treatment cycles, a recovery period that is not conducive to normal social interaction, and high prices. In response to skin photoaging, there is an increasing amount of research on improving mitochondrial function. Mitochondrial photobiological effects can improve mitochondrial function, reduce oxidative stress, reduce inflammatory responses, increase the number of mitochondria, and promote ATP production, thereby promoting the repair of photoaged cells and being an important treatment for skin photoaging. However, the complex chronic oxidative-inflammatory stress microenvironment of skin photoaging requires active drugs with multiple functional effects. The development of a new treatment method is crucial for the repair of photoaged skin.

[0004] Currently, there is considerable interest both domestically and internationally in the pharmaceutical value of the natural polyphenol chlorogenic acid and its potential to improve mitochondrial function and combat skin photoaging. However, most current natural polyphenolic active substances suffer from chemical instability, limited lipid solubility, poor transdermal penetration, low bioavailability, and susceptibility to esterase degradation, making their practical use difficult.

[0005] Therefore, there is an urgent need to solve the problems of chemical stability of multifunctional vitality drugs and their efficient transdermal delivery, so as to construct hydrogels for transdermal delivery of natural vitality drugs that continuously respond to human physiological temperature, providing a new method for solving skin photoaging. Summary of the Invention

[0006] In order to overcome the above-mentioned shortcomings and deficiencies of the prior art, the primary purpose of the present invention is to provide an application of chlorogenic acid and blue light in the preparation of anti-photoaging products.

[0007] Natural polyphenol chlorogenic acid has excellent biocompatibility, promotes collagen synthesis, resists skin photoaging, anti-oxidation, anti-inflammatory, and the ability to scavenge free radicals. Combining it with blue light and utilizing photobiological effects can achieve excellent photoaging effects.

[0008] Another object of the present invention is to provide a method for preparing a product that promotes adenosine triphosphate (ATP) production by combining chlorogenic acid and blue light. The present invention has found that chlorogenic acid has a mitochondrial photobiological effect under the action of blue light, which can promote ATP production.

[0009] Another object of the present invention is to provide a composite material loaded with chlorogenic acid according to the above application, specifically a thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid.

[0010] The chlorogenic acid-loaded thermosensitive transdermal release hydrogel of the present invention has excellent biocompatibility, the ability to scavenge oxygen free radicals, anti-oxidation, anti-inflammatory, and promote ATP production. It greatly enhances the bioavailability of chlorogenic acid through long-term thermosensitive transdermal release on the skin surface. Combined with the mitochondrial photobiological effect, it can repair the mitochondrial function of fibroblasts from multiple aspects and deep levels, promote mitochondrial ATP production, promote the repair of photoaged cells, achieve the effect of promoting the repair of photoaged skin, and achieve the purpose of treating skin photoaging.

[0011] Another object of the present invention is to provide a method for preparing the composite material.

[0012] Another object of the present invention is to provide the use of the above-mentioned composite material in the preparation of products for preventing / treating anti-photoaging, or in the preparation of skin tissue regeneration drugs, or in the preparation of tissue engineering materials (such as hydrogel dressings, especially medical dressings for resisting skin photoaging).

[0013] Another object of the present invention is to provide a method for inhibiting photoaging, comprising applying chlorogenic acid, or a composition containing chlorogenic acid, or a composite material loaded with chlorogenic acid to the skin, and then irradiating the skin with blue light; or applying a thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid to the skin.

[0014] The purpose of the present invention is achieved through the following solutions:

[0015] In a first aspect, the present invention provides an application of chlorogenic acid and blue light in combination, which is one of the following applications:

[0016] (1) Application in the preparation of anti-photoaging products;

[0017] (2) Application in the preparation of products that promote the production of adenosine triphosphate.

[0018] Furthermore, the product includes at least one of a medicine, a dressing, a device, a test kit, a chip, and an auxiliary health product.

[0019] The application is specifically carried out by applying chlorogenic acid, or a composition containing chlorogenic acid, or a composite material loaded with chlorogenic acid on the skin, and then irradiating with blue light.

[0020] Furthermore, the wavelength of the blue light is preferably 400-480 nm.

[0021] Furthermore, the blue light irradiation may have a dose of 1-800 mJ / cm 2 , preferably 300-500mJ / cm 2 .

[0022] Furthermore, the blue light irradiation may have a dose of 400±50 mJ / cm 2 .

[0023] Furthermore, the chlorogenic acid-loaded composite material preferably has temperature responsiveness, more preferably has temperature responsiveness to the physiological temperature of the human body.

[0024] The present invention has found that chlorogenic acid has a mitochondrial photobiological effect. Specifically, chlorogenic acid can react with reactive oxygen species in mitochondria to generate 440-540nm blue fluorescent light energy (FLE) under a specific wavelength of 400-480nm, thereby activating the mitochondrial respiratory chain and promoting the generation of adenosine triphosphate (ATP) in mitochondria, with the increase reaching 100%.

[0025] Based on the mechanism that chlorogenic acid can combine with the superoxide anion reaction products in cell mitochondria and produce blue fluorescence under blue light irradiation, FLE excitation can promote the mitochondria in cells irradiated by UVA to produce a large amount of ATP through photobiological regulation, reduce mitochondrial membrane potential, and accelerate cell repair.

[0026] The chlorogenic acid with mitochondrial photobiological effects described in the present invention has the abilities of anti-inflammatory, anti-oxidation, promoting collagen synthesis, targeting mitochondria to produce blue fluorescence, promoting ATP production, reducing the expression of aging-related galactosidase, etc., and can promote the repair of photoaged skin. It can be used in the preparation of anti-photoaging products or in the preparation of products that promote the production of adenosine triphosphate.

[0027] In a second aspect, the present invention provides a composite material loaded with chlorogenic acid according to the above application, specifically a thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid, which is prepared from the following raw materials: liposomes encapsulating chlorogenic acid, modified natural polysaccharide materials, modified gelatin and water.

[0028] Furthermore, the above-mentioned composite material can be prepared by conventional methods in the art, such as heating to mix the components evenly and then cooling to form a gel. The heating temperature can be the conventional heating temperature for preparing hydrogels in the art, such as 40-50°C. Cooling is preferably performed at 4°C for 1-3 hours. The composite material of the present invention is based on the cross-linking of modified natural polysaccharide materials and modified gelatin materials to form a stable hydrogel. The cross-linking mechanism of the hydrogel may include anionic and cationic cross-linking, intertwined cross-linking of polymer chains, crystallization cross-linking, dynamic reversible covalent cross-linking, hydrogen bond cross-linking, or a mixture of the above mechanisms.

[0029] Furthermore, the chlorogenic acid-encapsulated liposomes are prepared by conventional methods, such as using phosphatidylcholine and cholesterol via thin film hydration and sonication. Specifically, the method includes dissolving phosphatidylcholine and cholesterol, preparing a thin film, hydrating the film, encapsulating the chlorogenic acid, and sonicating the film to form the liposomes. The mass ratio of phosphatidylcholine to cholesterol can be 1:1 to 10:1. The solvent used for the hydration can be ether solution or water.

[0030] Furthermore, the content of chlorogenic acid in the chlorogenic acid-encapsulated liposome is 0.7-56 mg / mL.

[0031] Furthermore, the liposome has a particle size of 50 to 200 nm and a zeta potential of -15 mV to -30 mV, and has excellent permeability and stability in the skin.

[0032] In the chlorogenic acid-encapsulated liposomes of the present invention, chlorogenic acid is encapsulated inside the liposomes as the released active component to prevent its rapid degradation in the body, thereby overcoming the problems of chlorogenic acid itself, such as limited fat solubility, susceptibility to degradation by esterases, and low transdermal permeability, and improving its bioavailability.

[0033] In the chlorogenic acid-loaded composite material of the present invention, the content of chlorogenic acid is 0.1-100 μM, preferably 1-20 μM.

[0034] In the chlorogenic acid-loaded composite material of the present invention, the content of the modified natural polysaccharide material may be 1-25 wt %, such as 1-15 wt %.

[0035] In the chlorogenic acid-loaded composite material of the present invention, the content of modified gelatin may be 1-20 wt%, such as 1-15 wt%.

[0036] In the chlorogenic acid-loaded composite material of the present invention, the mass ratio of the modified natural polysaccharide material to the modified gelatin can be 0.5:1-2:1.

[0037] Furthermore, the natural polysaccharide material includes at least one of dextran, hyaluronic acid, chitin, and cellulose.

[0038] Furthermore, the above-mentioned modification refers to grafting modification of natural polysaccharide materials or gelatin using the same or different grafting compounds.

[0039] Furthermore, the grafted compound includes at least one of dopamine hydrochloride, caffeic acid, and phenylboric acid.

[0040] Furthermore, the molar ratio of the grafted compound to the natural polysaccharide material is 1:0.5-1:4.

[0041] Furthermore, the mass ratio of the grafted compound to gelatin is 1:0.5-1:10.

[0042] Furthermore, the modification of the natural polysaccharide material may include the following specific steps: firstly oxidizing the natural polysaccharide material with an oxidant, and then reacting the natural polysaccharide material with a grafting compound under the action of an activator to obtain a grafted modified natural polysaccharide material.

[0043] Furthermore, the gelatin is modified by reacting with a grafting compound under the action of an activator.

[0044] Furthermore, the oxidant may include at least one of sodium periodate, chromic acid and its salts (such as potassium dichromate, K2Cr2O7), potassium permanganate, manganese dioxide, nitric acid, etc. The molar ratio of the amount of the oxidant used to the natural polysaccharide material may be 1:0.1-1:4.

[0045] Furthermore, the oxidation of the natural polysaccharide material using an oxidant can be achieved by mixing the oxidant and the natural polysaccharide material in water, terminating the oxidation reaction with a terminator, and separating the oxidized natural polysaccharide material.

[0046] The concentration of the natural polysaccharide material in the oxidation reaction system can be 0.1-10% (w / v). The oxidation reaction time can be 6-12 hours. Conventional terminators, such as ethylene glycol and diethylene glycol, can be used. Conventional terminators can be used, such as a molar ratio of terminator to oxidant of 1:1-4:1. Separation can be performed by conventional methods such as dialysis and freeze-drying.

[0047] Furthermore, the activator may include at least one of N-hydroxysuccinimide / 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC / NHS), triphenylphosphine / diethyl azodicarboxylate (TPP / DEAD), 4-dimethylaminopyridine (DMAP), etc. The molar ratio of the activator to the grafted compound may be 1:0.5-1:4.

[0048] Furthermore, the grafting reaction is carried out in water. Specifically, the oxidized natural polysaccharide material / gelatin, the activator, and the grafting compound are added to water and stirred for reaction. The grafting reaction time can be 3-48 hours.

[0049] The present invention introduces chlorogenic acid-encapsulated liposomes into the field of tissue repair for the first time. The chlorogenic acid-loaded thermosensitive transdermal delivery hydrogel of the present invention is natural, mild, and highly efficient, and has a temperature responsiveness close to the physiological temperature of the human body. When applied to the skin, the hydrogel remains in a semi-gel state at the physiological temperature of the human body, and slowly releases the chlorogenic acid encapsulated by the liposomes, and penetrates into the skin cells through the skin barrier. The transdermal release of chlorogenic acid targets mitochondria, repairs and increases the number of mitochondria, promotes ATP production and cell regeneration and repair, thereby achieving excellent anti-photoaging effects.

[0050] The chlorogenic acid-loaded thermosensitive transdermal delivery hydrogel of the present invention can enhance the transdermal permeability of chlorogenic acid, promote its targeting to mitochondria in skin fibroblasts, and then generate fluorescent light energy (FLE) through photobiological regulation to promote mitochondrial repair, thereby effectively improving skin photoaging.

[0051] The chlorogenic acid-loaded thermosensitive transdermal delivery hydrogel of the present invention can further adjust the chlorogenic acid release rate by adjusting the ratio of the grafted compound to the natural polysaccharide material and gelatin. When the natural polysaccharide material and gelatin content is as low as 1 wt%, the composite material behaves as an ointment while still exhibiting excellent chlorogenic acid release and transdermal permeability.

[0052] The chlorogenic acid-loaded thermosensitive transdermal release hydrogel of the present invention has excellent biocompatibility, the ability to scavenge oxygen free radicals, anti-oxidation, anti-inflammatory, anti-skin photoaging, promote collagen synthesis, and promote ATP generation. It greatly enhances the bioavailability of chlorogenic acid through long-term thermosensitive transdermal release on the skin surface. Combined with the mitochondrial photobiological effect, it can repair the mitochondrial function of fibroblasts from multiple aspects and deep levels, promote mitochondrial ATP generation, promote photoaged cell repair, achieve the effect of promoting photoaged skin repair, and achieve the purpose of treating skin photoaging.

[0053] Compared with existing multifunctional bioactive substances: the chlorogenic acid-encapsulated liposomes of the present invention are simple and easy to implement, have the advantages of wide sources, low price, and the ability to target mitochondria to produce mitochondrial photobiological effects; the chlorogenic acid-loaded thermosensitive transdermal delivery hydrogel of the present invention has good biocompatibility, and combining it with the mitochondrial photobiological effects of chlorogenic acid and applying it in the biomedical field, such as in the preparation of tissue engineering materials, such as hydrogel dressings, especially in the preparation of medical dressings for anti-skin photoaging, and in the preparation of skin tissue regeneration drugs or dressings, has significant advantages.

[0054] In a third aspect, the present invention further provides the use of the above-mentioned chlorogenic acid-loaded composite material in the preparation of products for preventing / treating anti-photoaging, or in the preparation of skin tissue repair / regeneration drugs, or in the preparation of tissue engineering materials (such as hydrogel dressings, especially medical dressings for resisting skin photoaging).

[0055] The chlorogenic acid-loaded thermosensitive sustained-release hydrogel of the present invention is used to prepare a dressing, which can effectively promote collagen synthesis, resist skin photoaging, anti-oxidation, anti-inflammation, and scavenging free radicals, improve the bioavailability of chlorogenic acid, promote its transdermal delivery and long-term release on the skin surface, and promote the repair of photoaged skin tissue.

[0056] In a fourth aspect, the present invention also provides a method for inhibiting photoaging, comprising applying chlorogenic acid, or a composition containing chlorogenic acid, or a composite material loaded with chlorogenic acid to the skin, and then irradiating the skin with blue light; or applying the thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid to the skin.

[0057] The present invention utilizes the mitochondrial photobiological effect of chlorogenic acid to activate and repair mitochondria in tissue cells by stimulating photobiological reactions, thereby increasing the number of mitochondria, promoting mitochondria to produce more ATP, reducing aging-related galactose and regulating immune cells, thereby achieving multiple effects such as anti-inflammatory, anti-oxidation, accelerated clearance of senescent cells, and stimulation of collagen secretion to promote cell regeneration and repair. It is an effective method for treating skin photoaging and has the significant advantages of wide source, low price, safety and environmental protection. BRIEF DESCRIPTION OF THE DRAWINGS

[0058] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0059] Figure 1 This is the UV-visible spectrum of chlorogenic acid liposomes.

[0060] Figure 2 This is a statistical graph showing the effect of chlorogenic acid on the viability of human fibroblasts.

[0061] Figure 3 This is a diagram of the blue fluorescence photobiological effect of chlorogenic acid on mitochondria.

[0062] Figure 4 This is a statistical graph showing the effect of chlorogenic acid on ATP production in human fibroblasts treated with UV.

[0063] Figure 5Statistical graphs of superoxide dismutase activity (A), catalase activity (B) and malondialdehyde content (C) in human fibroblasts after ultraviolet treatment with chlorogenic acid combined with mitochondrial photobiological effect.

[0064] Figure 6 This is the senescence-related galactosidase staining of human fibroblasts after ultraviolet treatment with chlorogenic acid combined with mitochondrial photobiological effect.

[0065] Figure 7 This is a statistical graph showing the effect of chlorogenic acid thermosensitive transdermal delivery hydrogel on human fibroblast viability.

[0066] Figure 8-Figure 9 This is the temperature-dependent rheological diagram of the thermosensitive transdermal delivery hydrogel of chlorogenic acid.

[0067] Figure 10 The transdermal delivery curve of chlorogenic acid thermosensitive transdermal delivery hydrogel (A) and the statistical chart of skin drug retention (B).

[0068] Figure 11 Fluorescence image of chlorogenic acid thermosensitive transdermal delivery hydrogel clearing active oxygen in human fibroblasts.

[0069] Figure 12 This is a picture of the photoaging repair effect on mouse skin.

[0070] Figure 13 Epidermal thickness (A), stratum corneum water content (B) and transepidermal water loss coefficient (C) of mouse skin.

[0071] Among them, *p<0.05, **p<0.01, ***p<0.001. DETAILED DESCRIPTION

[0072] The present invention is further described in detail below with reference to the following examples, but the embodiments of the present invention are not limited thereto. Materials used in the following examples are commercially available unless otherwise specified. Methods described are conventional methods unless otherwise specified. The amounts of each component are expressed in parts by mass, parts by volume, and parts by mole (g, mL, mol).

[0073] The chlorogenic acid-loaded composite material used in the embodiments of the present invention is specifically a chlorogenic acid-loaded thermosensitive transdermal delivery hydrogel, which is prepared from the following raw materials: chlorogenic acid-loaded liposomes, modified natural polysaccharide materials, modified gelatin, and water.

[0074] In a specific embodiment of the present invention, a thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid can be prepared by mixing the components, heating at 40-50°C to achieve uniform mixing, and then cooling at 4°C for 1-3 hours to form a gel. The composite material of the present invention forms a stable hydrogel based on the crosslinking of a modified natural polysaccharide material with a modified gelatin material. The crosslinking mechanism of the hydrogel may include cationic and anionic crosslinking, intertwining crosslinking of polymer chains, crystallization crosslinking, dynamic reversible covalent crosslinking, hydrogen bonding, or a combination of these mechanisms.

[0075] In one embodiment, the content of chlorogenic acid is 0.1-100 μM; in another embodiment, the content of chlorogenic acid is 1-20 μM.

[0076] In one embodiment, the content of the modified natural polysaccharide material is 1-25 wt %; in another embodiment, the content of the modified natural polysaccharide material is 1-15 wt %.

[0077] In one embodiment, the content of the modified gelatin is 1-20 wt %; in another embodiment, the content of the modified natural polysaccharide material is 1-15 wt %.

[0078] In one embodiment, the mass ratio of the modified natural polysaccharide material to the modified gelatin is 0.5:1-2:1; in another embodiment, the mass ratio of the modified natural polysaccharide material to the modified gelatin is 1:1.

[0079] The chlorogenic acid-loaded liposomes used in the embodiments of the present invention can be prepared by conventional methods, specifically by the following steps:

[0080] L-α-phosphatidylcholine and cholesterol are mixed in a mass ratio of 1:1-10:1, and then a chloroform solution is added and magnetically stirred until uniformly dissolved; vacuum rotary evaporation is performed at 30-50°C until the chloroform is completely evaporated to form a uniform film, and then rotary drying is continued for 20-40 minutes; ether is added and stirred at room temperature until the film is completely dissolved, 1-80 mg / mL chlorogenic acid aqueous solution is added with an oil-water ratio of 2:1-5:1, and ultrasonic cell crusher is used for ice bath ultrasonic treatment for 10-20 minutes, vacuum rotary evaporation is performed at 30-50°C until the ether is completely evaporated, hydration is carried out for 1-3 hours, and then ultrasonic treatment is performed in an ice bath with an ultrasonic cell crusher for 10-20 minutes to obtain chlorogenic acid-loaded liposomes, wherein the content of chlorogenic acid in the liposomes is 0.7-56 mg / mL.

[0081] In one embodiment, the liposome has a particle size of 50 to 200 nm and a zeta potential of -15 mV to -30 mV, and has excellent permeability and stability in the skin.

[0082] In one embodiment, 10 μg / mL chlorogenic acid liposomes were prepared with deionized water and subjected to UV-visible spectroscopy. The absorption spectrum was as follows: Figure 1 As shown, it can be seen that its maximum absorption peak appears at 324 nm, A(324 nm)=0.56.

[0083] The modified natural polysaccharide material and modified gelatin used in the embodiments of the present invention are obtained by grafting and modifying with the same or different grafting compounds.

[0084] In one embodiment, the modified natural polysaccharide material is prepared by the following steps: first, 1 mole of an oxidant and 0.1-4 moles of a natural polysaccharide material are added to water and mixed to react for 6-12 hours, 1-4 moles of a terminator (ethylene glycol, diethylene glycol, etc.) are added to terminate the oxidation reaction, and the oxidized natural polysaccharide material is separated; 0.5-4 moles of the oxidized natural polysaccharide material is added to water, and reacted with 1 mole of a grafted compound under the action of 0.5-4 moles of an activator (EDC / NHS, TPP / DEAD or DMAP) for 3-48 hours to obtain a grafted modified natural polysaccharide material.

[0085] In a specific embodiment of the present invention, the oxidant may include at least one of sodium periodate, chromic acid and its salts (such as potassium dichromate, K2Cr2O7), potassium permanganate, manganese dioxide, nitric acid, etc.

[0086] In a specific embodiment of the present invention, the natural polysaccharide material includes at least one of dextran, hyaluronic acid, chitin, and cellulose.

[0087] In a specific embodiment of the present invention, the grafted compound includes at least one of dopamine hydrochloride, caffeic acid, and phenylboronic acid.

[0088] In a specific embodiment of the present invention, the concentration of the natural polysaccharide material in the oxidation reaction system may be 0.1-10% (w / v).

[0089] In a specific embodiment of the present invention, the grafting reaction is carried out in a closed manner under the protection of an inert gas.

[0090] In a specific embodiment of the present invention, the pH of the grafting reaction system is preferably weakly acidic, preferably 4.5-5.5. For example, the pH can be adjusted by conventional acidic reagents such as hydrochloric acid and sodium hydroxide (e.g., 1 M hydrochloric acid and sodium hydroxide).

[0091] In a specific embodiment of the present invention, after the grafting reaction is completed, the grafted modified natural polysaccharide material can be separated by conventional means such as dialysis and freeze drying. The dialysis is preferably performed in a weakly acidic environment.

[0092] In one embodiment, modified gelatin is prepared by adding gelatin to water and reacting it with 1 mole of a grafting compound in the presence of 0.5-4 moles of an activator (EDC / NHS, TPP / DEAD, or DMAP) at 35-55°C for 3-48 hours to obtain a grafted modified natural polysaccharide material. The mass ratio of the grafting compound to gelatin is 1:0.5-1:10.

[0093] In a specific embodiment of the present invention, the concentration of gelatin in the reaction system is 1-20 wt%.

[0094] In a specific embodiment of the present invention, the mass ratio of the grafted compound to gelatin is 1:0.1-1:20.

[0095] In a specific embodiment of the present invention, after the grafting reaction is completed, the grafted modified gelatin material can be separated by conventional means such as dialysis and freeze drying. The dialysis is preferably performed at 37-45°C.

[0096] In one embodiment, the application of chlorogenic acid and blue light in combination is one of the following applications:

[0097] (1) Application in the preparation of anti-photoaging products;

[0098] (2) Application in the preparation of products that promote the production of adenosine triphosphate.

[0099] In a specific embodiment of the present invention, the product includes at least one of a medicine, a dressing, a device, a test kit, a chip, and an auxiliary health product.

[0100] In a specific embodiment of the present invention, the application can be achieved by applying chlorogenic acid, or a composition containing chlorogenic acid, or a composite material loaded with chlorogenic acid on the skin, and then irradiating with blue light.

[0101] In a specific embodiment of the present invention, the application can be achieved by applying chlorogenic acid-encapsulated liposomes, or a composition containing chlorogenic acid, or a composite material loaded with chlorogenic acid on the skin, and then irradiating with blue light.

[0102] In another embodiment, the composite material loaded with chlorogenic acid is used in the preparation of products for preventing / treating anti-photoaging, or in the preparation of skin tissue repair / regeneration drugs, or in the preparation of tissue engineering materials (such as hydrogel dressings, especially medical dressings for resisting skin photoaging).

[0103] In a specific embodiment of the present invention, the drug further comprises one or more pharmaceutically acceptable carriers or excipients. The pharmaceutically acceptable carriers or excipients may contain inert ingredients that do not unduly inhibit the biological activity of the compound. Pharmaceutically acceptable carriers or excipients should be biocompatible, for example, non-toxic, non-inflammatory, non-immunogenic or without other undesirable reactions or side effects when administered to a subject. Standard pharmaceutical formulation techniques can be used.

[0104] In a specific embodiment of the present invention, pharmaceutically acceptable carriers or excipients include, but are not limited to, diluents, binders, surfactants, wetting agents, adsorption carriers, lubricants, fillers, disintegrants, preservatives, etc. These substances are used to help the stability of the formulation, or to help improve the activity or its biological effectiveness, or to produce an acceptable taste or smell in the case of oral administration. The preparations that can be used in such pharmaceutical compositions can be in the form of the original compound itself or optionally in the form of a pharmaceutically acceptable salt thereof. The pharmaceutical composition thus formulated can be administered by any appropriate method known to those skilled in the art as needed.

[0105] In a specific embodiment of the present invention, the diluent includes but is not limited to lactose, sodium chloride, glucose, urea, starch, water.

[0106] In a particular embodiment of the present invention, binders include, but are not limited to, starch, pregelatinized starch, dextrin, maltodextrin, sucrose, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, ethylcellulose, polyvinyl alcohol, polyethylene glycol, polyvinylpyrrolidone, alginic acid and alginates, xanthan gum, hydroxypropyl cellulose and hydroxypropyl methylcellulose.

[0107] In a specific embodiment of the present invention, the surfactant includes but is not limited to polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, stearic acid monoglyceride, and cetyl alcohol.

[0108] In a specific embodiment of the present invention, the humectant includes but is not limited to glycerin and starch.

[0109] In a specific embodiment of the present invention, the adsorption carrier includes, but is not limited to, starch, lactose, bentonite, silica gel, kaolin and bentonite.

[0110] In a specific embodiment of the present invention, lubricants include but are not limited to zinc stearate, glyceryl monostearate, polyethylene glycol, talc, calcium and magnesium stearate, polyethylene glycol, boric acid powder, hydrogenated vegetable oil, sodium stearyl fumarate, polyoxyethylene monostearate, monolauric sucrose ester, sodium lauryl sulfate, magnesium lauryl sulfate, and magnesium lauryl sulfate.

[0111] In a specific embodiment of the present invention, fillers include, but are not limited to, mannitol (granular or powdered), xylitol, sorbitol, maltose, erythrose, microcrystalline cellulose, polymeric sugars, coupling sugars, glucose, lactose, sucrose, dextrin, starch, sodium alginate, laminarin powder, agar powder, calcium carbonate, and sodium bicarbonate.

[0112] In a specific embodiment of the present invention, the disintegrant includes but is not limited to cross-linked vinyl pyrrolidone, sodium carboxymethyl starch, low-substituted hydroxypropyl methyl, cross-linked sodium carboxymethyl cellulose, and soybean polysaccharide.

[0113] In the following implementation, the UV treatment time is calculated based on the required dose and the irradiation intensity of the lamp.

[0114] Example 1: Combination of chlorogenic acid and blue light to promote ATP production

[0115] (1) The chlorogenic acid content in the cell culture medium was adjusted to obtain four culture mediums with different chlorogenic acid contents. The final concentrations of chlorogenic acid were 1, 5, 10, and 20 μM, respectively. A group of cell culture medium without chlorogenic acid was set as a control group for cell culture experiments. The above five groups of culture medium were co-cultured with human fibroblasts. The five groups of cells were placed in a constant temperature incubator at 37°C containing 5% CO2. The cell proliferation was detected using a CCK-8 kit on the 1st and 2nd day of culture. The results are as follows: Figure 2 shown.

[0116] Depend on Figure 2 As shown in the cell viability statistics, chlorogenic acid can promote fibroblast proliferation at certain concentrations. When the chlorogenic acid content in the culture medium reaches 10 μM, the cell viability remains at 100% after 2 days of co-culture, indicating that the chlorogenic acid of the present invention is not cytotoxic at concentrations of 1-10 μM. However, when the chlorogenic acid content in the culture medium reaches 20 μM, the cell viability is only 87% after 1 day of co-culture, and only 83% after 2 days of co-culture, indicating that the chlorogenic acid of the present invention is cytotoxic at a concentration of 20 μM.

[0117] (2) Human fibroblasts treated with UV light were used to evaluate the mitochondrial photobiological effect of chlorogenic acid. Based on the above cell viability results, fibroblasts were treated with 10 μM chlorogenic acid for 1 h and 18 J / cm 2 Human fibroblasts were treated with UVA, and then treated with 10 μM chlorogenic acid for 30 minutes. The culture medium was removed and the cells were stained with JC-1 dye for 30 minutes. The culture medium was replaced with ordinary culture medium. The cells were placed on a laser confocal microscope to observe the targeting of chlorogenic acid to mitochondria and its photobiological effects. The results are shown in the figure. Figure 3 As shown in the figure, chlorogenic acid can target mitochondria and produce fluorescence under blue light irradiation in a confocal microscope.

[0118] (3) Enhanced ATP detection kit was used to detect the changes in ATP in human fibroblasts after treatment. According to the treatment conditions, the cells were divided into UV group, chlorogenic acid group, and chlorogenic acid fluorescence energy group. The control group was treated with 18 J / cm 2 UVA was used to treat cells; 18 J / cm 2 UVA cells were then treated with 10 μM chlorogenic acid for 30 min; the chlorogenic acid fluorescence energy group was treated with 18 J / cm 2 Cells were treated with UVA, then treated with 10 μM chlorogenic acid for 30 min, and then irradiated with 400-480 nm blue light for 10 min at a dose of 400 mJ / cm 2 After the four groups of cells were replaced with ordinary culture medium, they were cultured in a constant temperature incubator at 37℃ with 5% CO2 for 24 hours. The enhanced ATP detection kit was used to detect the changes in ATP in fibroblasts. The results are shown in Figure 4 As shown in the figure, the ATP content of cells in the UV group decreased, while the ATP content of cells in the chlorogenic acid group and the chlorogenic acid fluorescence light energy group increased. In particular, the chlorogenic acid fluorescence light energy group significantly increased the production of ATP in cells, which had a repairing effect on the damage caused by UV irradiation.

[0119] Example 2: Combination of chlorogenic acid and blue light enhances the anti-photoaging effect of human fibroblasts

[0120] Human fibroblasts treated with ultraviolet light were used to evaluate the anti-cell photoaging effect of chlorogenic acid combined with mitochondrial photobiological effects and blue light. 2 Human fibroblasts were treated with UVA, treated with 10μM chlorogenic acid for 30 minutes, and then irradiated with 400-480nm blue light for 10 minutes. The cells were placed in a 37℃ constant temperature incubator containing 5% CO2 for 24 hours, and the culture medium was removed to characterize the cells' resistance to light aging. First, the cells were stained with senescence-associated galactosidase (SA-β-gal) dye for 12 hours, washed with PBS three times, and the cells were photographed under a microscope to observe the expression of the cell senescence marker SA-β-gal. Secondly, the superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) of the cells were quantitatively analyzed. The results are shown in Figure 5-Figure 6 .

[0121] Depend on Figure 5 It can be seen that compared with chlorogenic acid alone, chlorogenic acid combined with mitochondrial photobiological effect and blue light treatment can significantly enhance the superoxide dismutase ( Figure 5 A) and catalase activity ( Figure 5 B), reducing the content of malondialdehyde, a lipid peroxidation product in human fibroblasts ( Figure 5 C) By Figure 6 It can be seen that compared with chlorogenic acid alone, chlorogenic acid combined with mitochondrial photobiological effects and blue light combined treatment can significantly reduce the expression of SA-β-gal in human fibroblasts. The results show that compared with chlorogenic acid alone, chlorogenic acid combined with mitochondrial photobiological effects and blue light combined treatment can significantly enhance the anti-photoaging effect of human fibroblasts.

[0122] Example 3: Preparation of chlorogenic acid-loaded composite material

[0123] (1) Chlorogenic acid-encapsulated liposomes were prepared by thin film hydration ultrasonic method. L-α-phosphatidylcholine and cholesterol were mixed in a mass ratio of 5:1 and added to chloroform solution with magnetic stirring until uniform dissolution. The mixture was vacuum evaporated in a rotary evaporator at 40°C until the chloroform was completely evaporated to form a uniform film. The film was then rotary dried for 30 minutes. Ether was added and stirred at room temperature until the film was completely dissolved. A chlorogenic acid aqueous solution was added with an oil-water ratio of 3.5:1. The mixture was ultrasonically treated in an ice bath for 15 minutes using an ultrasonic cell crusher. The mixture was vacuum evaporated in a rotary evaporator at 40°C until the ether was completely evaporated. The mixture was hydrated for 2 hours and then ultrasonically treated in an ice bath for 15 minutes using an ultrasonic cell crusher (3s on, 3s off) to obtain chlorogenic acid-encapsulated liposomes. The content of chlorogenic acid in the liposomes was 0.7-56 mg / mL.

[0124] (2) Add 5 parts by mass of hyaluronic acid to 500 parts by volume of water, stir thoroughly, add 1-1.5 parts by mass of sodium periodate, react in the dark for 6-12 hours, and then add ethylene glycol to terminate the reaction. The solution after the oxidation reaction is dialyzed under a neutral environment for 72 hours and freeze-dried to obtain oxidized hyaluronic acid (OHA);

[0125] OHA is added to water, followed by 1-2 parts by weight of EDC and 1-2 parts by weight of NHS. The pH is adjusted to 4.6-4.8 with 1M HCl solution and stirred at room temperature for 30 minutes. Dopamine hydrochloride is then added to the solution, and the pH is adjusted to 4.6-4.8 with 1M HCl solution. The reaction is stirred in the dark under nitrogen for 12 hours. The solution is then transferred to a dialysis bag and dialyzed against deionized water at pH 4.6 for 2 days and then against deionized water at pH 5.5 for 1 day, with the water changed three times daily. Finally, the dialysate is freeze-dried to obtain dopamine-grafted oxidized hyaluronic acid (ODA). The molar ratio of the grafted compound to the natural polysaccharide material can be 1:0.5-1:4. For better comparison, the molar ratio of OHA to dopamine hydrochloride used in the gel preparation of the following examples is 1:1.

[0126] (3) Add 10 parts by mass of gelatin (Gel) to 100 parts by volume of water, stir thoroughly at 40°C, add 1-2 parts by mass of EDC and 1-2 parts by mass of NHS, adjust the pH to 5.0-6.0 with 1M HCl solution, stir at room temperature for 30 minutes, add phenylboric acid to the solution, react for 24 hours, then transfer the solution to a dialysis bag and dialyze with deionized water at 40°C for 3 days, changing the water three times a day. Finally, the dialysate is freeze-dried to obtain modified gelatin grafted with phenylboronic acid (GBA). The mass ratio of the grafted compound used to the gelatin is 1:0.5-1:10. For better comparison, the mass ratio of phenylboric acid to gelatin used in the gel preparation of the following example products is 1:5.

[0127] (4) Add chlorogenic acid-encapsulated liposomes and modified gelatin to water, mix them evenly, and then mix them with ODA. After heating at 40-50°C to mix evenly, cool them at 4°C for 1-3 hours to form a gel, thereby obtaining a thermosensitive hydrogel encapsulating chlorogenic acid-encapsulated liposomes. The content of modified gelatin in the hydrogel can be 1-25 wt%; the content of ODA can be 1-20 wt%; and the content of chlorogenic acid can be 0.1-100 μM, which can be adjusted according to the amount of liposomes used.

[0128] (5) Temperature responsiveness rheological property test of thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid: A rotational rheometer was used to perform temperature scanning on hydrogels with a chlorogenic acid content of 10 μM modified gelatin and a final ODA content of 6 wt% to evaluate their temperature responsiveness. The hydrogel sample (10 mm in diameter, 1 mm in thickness) was placed between 20 mm parallel plates and subjected to a temperature scanning test from 0 to 50.5 °C at a constant strain of 1%, a constant frequency of 10 rad / s, and a heating rate of 3 °C / min. The changes in the storage modulus (G') and loss modulus (G") of the hydrogel were recorded to study the temperature responsiveness of the hydrogel; gelatin gel with the same content (12 wt%) was used as a comparison, and the results are shown in FIG. Figure 8 .Depend on Figure 8 As shown in A, the gelation transition temperature of gelatin gel is 31.9°C, while the gelation transition temperature of the thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid of the present invention is 38.6°C ( Figure 8 B), which indicates that it can maintain a semi-gel state and release drugs in a temperature-responsive manner at human physiological temperature.

[0129] Adjust the content of ODA and GBA in the hydrogel. When the content is 1wt% and 1wt% respectively, the gelation transition temperature is 36.2℃( Figure 9 A); when its content is 6wt% and 12wt%, the gelation transition temperature is 41.1℃( Figure 9B) That is, the thermosensitive hydrogel containing chlorogenic acid liposomes of the present invention has similar gelation transition temperatures when the contents of ODA and GBA are similar, and can maintain a semi-gel state at the physiological temperature of the human body to release drugs in a temperature-responsive manner.

[0130] Example 4: Cytotoxicity of chlorogenic acid-loaded composite materials

[0131] The preparation of the composite material loaded with chlorogenic acid was the same as in Example 3. For better comparison, the final contents of modified gelatin and ODA in the following examples were 6 wt %, respectively. The content of chlorogenic acid in the hydrogel was adjusted to obtain five hydrogels with different chlorogenic acid contents. 0.1, 0.5, 1 and 2 μL of 0.7 mg / mL chlorogenic acid liposomes were added to 200 μL of the above hydrogel precursor solution (the final content of modified gelatin and ODA was 6 wt %, respectively) to prepare chlorogenic acid thermosensitive hydrogels with chlorogenic acid contents of 0, 1, 5, 10, and 20 μM (the hydrogel group was 0 μM chlorogenic acid thermosensitive hydrogel). A group of well plates without hydrogels were set up as a control group for cell culture experiments. Transwell chambers were used to co-culture with human fibroblasts. After the cells were seeded in the well plates, the Transwell chambers were covered, and 200 μL of the hydrogel prepared by the precursor solution was placed in the chamber, and 800 μL of culture medium was added. The six groups of cells were co-cultured in a constant temperature incubator at 37°C with 5% CO2 for 1 day, and the cell proliferation was detected using a CCK-8 kit. The results are as follows. Figure 7 shown.

[0132] Depend on Figure 7 From the cell viability statistics, it can be seen that when the content of chlorogenic acid in the hydrogel reaches 20 μM, the cell viability is still 95% after one day of co-culture with the hydrogel and cells, which is higher than the cell activity (87%) when 20 μM chlorogenic acid is directly used. This shows that chlorogenic acid can be encapsulated in the hydrogel to achieve almost no cytotoxicity when the chlorogenic acid concentration is as high as 20 μM. The hydrogel of the present invention can effectively increase the effective use concentration of chlorogenic acid.

[0133] Example 5: In vitro transdermal release test of thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid

[0134] The transdermal permeation of liposomes in liposome-hydrogel was tested using a transdermal diffusion tester. The effective permeation area of ​​the Franz diffusion cell was 1.2 cm 2The receptor was a mixture of 8 mL of PBS / ethanol (pH 6.0, 3:1, v / v), stirred with a magnetic stirrer at 300 rpm in a 37°C water bath. The drug content in the receptor was measured using a UV spectrophotometer. At predetermined time points (0.5, 1, 2, 4, 8, 12, and 24 h), 2 mL of the receptor solution was taken to determine the amount of drug permeating the Bama Xiang pig skin, and an equal volume of PBS was added. After 24 h of transdermal penetration, the Bama Xiang pig skin was washed and clamped. Then, the skin was extracted with water ultrasound for 1 h, the supernatant was collected, and the drug skin retention was determined. The results are shown in Table 1. Figure 10 The chlorogenic acid liposomes were directly applied, and the chlorogenic acid-loaded hydrogels prepared with the same content of modified hyaluronic acid and unmodified gelatin (the note in the figure is the single modified natural polysaccharide chlorogenic acid hydrogel) were compared. The chlorogenic acid content was 10μM in all cases.

[0135] As can be seen from the figure, the effective transdermal concentration of the thermosensitive transdermal delivery hydrogel of chlorogenic acid (chlorogenic acid hydrogel) of the present invention is 18.7 μg / cm 2 , with the best transdermal release effect; compared with chlorogenic acid liposomes and single-modified natural polysaccharide chlorogenic acid hydrogels, the chlorogenic acid thermosensitive transdermal delivery hydrogel of the present invention can promote the transdermal delivery of chlorogenic acid. Moreover, compared with chlorogenic acid liposomes and single-modified natural polysaccharide chlorogenic acid hydrogels, the chlorogenic acid thermosensitive transdermal delivery hydrogel of the present invention can increase the retention of chlorogenic acid in the skin.

[0136] Example 6: Antioxidant performance test of thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid liposomes

[0137] The cell viability oxygen detection kit was used to analyze the effect of chlorogenic acid on the clearance of active oxygen in human fibroblasts. First, 0.5 mL of cell suspension (4 × 10 4 Cells / mL), after culturing for 24 hours, the culture medium was aspirated, and 0.5 mL of culture medium containing H2O2 (100 μM) was added to each well to continue culturing the cells for 3 hours to allow the cells to produce oxidative stress. The culture medium was aspirated, and the cells were treated with 1, 5, 10 and 20 μM chlorogenic acid hydrogel for 2 days, the culture medium was aspirated, the cells were washed once with PBS, DCFH-DA dye solution was added, and the well plate was placed in a cell culture incubator for incubation in the dark for 30 minutes. After washing the cells with PBS, DAPI dye solution was added and stained at room temperature in the dark for 5 minutes. The dye solution was aspirated, the cells were washed with PBS, the PBS was aspirated, and the well plate was placed under a fluorescence microscope for observation and photography. Among them, the negative control group did not add any material and did not receive any treatment (no oxidative stress treatment), and the positive control group cells were directly cultured with normal culture medium after oxidative stress (no treatment). The results are shown in the figure. Figure 11 .

[0138] As shown in the figure, the thermosensitive transdermal delivery hydrogel of chlorogenic acid of the present invention has excellent antioxidant capacity and can effectively remove active oxygen in cells, and the removal effect has a dose-effect relationship.

[0139] Example 7: Thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid liposomes combined with mitochondrial fluorescence regeneration technology for the treatment of skin photoaging

[0140] Thirty female C57 mice (6-8 weeks old) were shaved and irradiated with UV light on the 2nd and 5th day of each week, with 1200 mJ / cm 2 UVA+200mJ / cm 2 Mice were treated with UVB once for 4 weeks to create an animal model of skin photoaging. The experimental group was divided into 5 groups: A: negative control group (normal mice); B: positive control group (no treatment after modeling); C: hydrogel group (thermosensitive hydrogel group with 0 chlorogenic acid, hydrogel specifications: length * width * height = 30mm * 30mm * 1mm, applied for 24 hours); D: chlorogenic acid hydrogel group (chlorogenic acid content is 10μM, hydrogel specifications: length * width * height = 30mm * 30mm * 1mm, applied for 24 hours); E: chlorogenic acid hydrogel + fluorescent light energy group (chlorogenic acid content is 10μM, hydrogel specifications: length * width * height = 30mm * 30mm * 1mm, after 24 hours of application, the hydrogel was removed and blue light irradiation was applied for 10 minutes; the dose was 400mj / cm 2 ). Take photos every day to observe whether the skin on the back of the mice shows signs of desquamation, pigmentation, thickening, leatheriness, edema, pigmentation, etc., and record the time of onset and duration. Before killing the mice after 4 weeks of light exposure, skin thickness measurement and skin barrier function test were performed. The treatment effect is as follows Figure 12 and Figure 13 As shown. Figure 12 It can be seen that the thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid liposomes of the present invention combined with the mitochondrial photobiological effect under blue light irradiation can effectively improve skin photoaging desquamation, erythema, dryness, roughness, pigmentation, etc. Figure 13 It can be seen that on the 28th day, the epidermal thickness, stratum corneum water content, and transepidermal water loss coefficient of mice after treatment with the thermosensitive transdermal delivery hydrogel of chlorogenic acid liposomes combined with the mitochondrial photobiological effect under blue light irradiation were closer to normal skin, and the effect of treating skin photoaging was the best.

[0141] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. The application of chlorogenic acid and blue light in the preparation of anti-photoaging products is characterized by: The blue light irradiation dose is 1-800mJ / cm 2 ; The wavelength of the blue light is 400-480nm; The application is carried out by applying chlorogenic acid or a thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid on the skin, and then implementing blue light irradiation.

2. The use according to claim 1, characterized in that: Application of chlorogenic acid and blue light in the preparation of products that promote the production of adenosine triphosphate.

3. The use according to claim 1 or 2, characterized in that The product includes at least one of medicine, dressing and auxiliary health care product.

4. The use according to claim 1, characterized in that The thermosensitive transdermal delivery hydrogel loaded with chlorogenic acid is prepared from the following raw materials: liposomes loaded with chlorogenic acid, modified natural polysaccharide materials, modified gelatin and water; The chlorogenic acid-loaded thermosensitive transdermal delivery hydrogel is prepared by mixing the components, heating at 40-50° C. to uniformly mix the components, and then cooling at 4° C. for 1-3 hours to form a gel. The content of the chlorogenic acid is 1-20 μM; the content of the modified natural polysaccharide material is 1-15 wt %; the mass ratio of the modified natural polysaccharide material to the modified gelatin is 0.5:1-2:1; The chlorogenic acid-loaded liposomes are prepared by the following steps: La-phosphatidylcholine and cholesterol are mixed in a mass ratio of 1:1-10:1, and then a chloroform solution is added and magnetically stirred until uniformly dissolved; vacuum rotary evaporation is performed at 30-50°C until the chloroform is completely evaporated to form a uniform film, and then rotary drying is continued for 20-40 minutes; ether is added and stirred at room temperature until the film is completely dissolved, 1-80 mg / mL chlorogenic acid aqueous solution is added with an oil-water ratio of 2:1-5:1, and ultrasonic cell crusher is used for ice bath ultrasonic treatment for 10-20 minutes, vacuum rotary evaporation is performed at 30-50°C until the ether is completely evaporated, hydration is performed for 1-3 hours, and then ultrasonic cell crusher is used for ice bath ultrasonic treatment for 10-20 minutes to obtain chlorogenic acid-loaded liposomes; The modified natural polysaccharide material is prepared by the following steps: first, adding 1 mole of sodium periodate and 0.1-4 moles of hyaluronic acid to water, mixing and reacting for 6-12 hours, adding 1-4 moles of ethylene glycol to terminate the oxidation reaction, and separating to obtain oxidized hyaluronic acid; adding 0.5-4 moles of oxidized hyaluronic acid to water, reacting with 1 mole of dopamine hydrochloride in the presence of 0.5-4 moles of an activator, EDC / NHS, for 3-48 hours to obtain oxidized hyaluronic acid grafted with dopamine; The modified gelatin is prepared by the following steps: adding gelatin to water, reacting with 1 mole of grafting compound at 35-55° C. for 3-48 hours under the action of 0.5-4 mole of activator EDC / NHS to obtain grafted modified gelatin; The mass ratio of the grafted compound used to gelatin is 1:0.5-1:10; The grafted compound is phenylboronic acid.

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