An iron death inhibitor ophthalmic delivery system based on cyclodextrin inclusion and liposome drug loading and a preparation method and application thereof

Abstract

This invention discloses an ophthalmic delivery system for ferroptosis inhibitors based on cyclodextrin inclusion and liposome loading, its preparation method, and its application, belonging to the field of biomedical technology. Addressing the poor water solubility of ferroptosis inhibitors, this invention first utilizes hydrophilic cyclodextrin to encapsulate them into water-soluble inclusion complexes, significantly improving their solubility and stability. Then, the inclusion complex is used as the inner aqueous phase and encapsulated in liposome vesicles composed of phospholipids and cholesterol, forming a dual drug delivery system of "inclusion complex-liposome". This delivery system combines the solubilizing and stabilizing properties of cyclodextrin with the bioadhesive, sustained-release, and permeation-enhancing advantages of liposomes, effectively prolonging the drug's residence time on the ocular surface, promoting transcorneal absorption, and targeting inflammatory lesions on the ocular surface, efficiently inhibiting ferroptosis, thereby achieving significant treatment for ocular surface inflammatory diseases such as dry eye and keratoconjunctivitis.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to an ophthalmic delivery system for ferroptosis inhibitors based on cyclodextrin inclusion and liposome loading, its preparation method and application. Background Technology

[0002] Ocular surface inflammation is a common ophthalmic pathological process affecting the cornea, conjunctiva, eyelid margin, and lacrimal glands, encompassing various diseases such as dry eye syndrome, allergic conjunctivitis, and infectious keratoconjunctivitis. Its typical characteristics include immune cell infiltration, elevated levels of pro-inflammatory cytokines (such as IL-1β, IL-6, and TNF-α) and matrix metalloproteinases (MMPs), ocular surface epithelial damage, and oxidative stress. Currently, first-line drugs for the clinical treatment of ocular surface inflammation mainly include corticosteroids (such as fluorometholone, dexamethasone, and prednisolone acetate) and nonsteroidal anti-inflammatory drugs (NSAIDs) (such as diclofenac sodium, bromfenac sodium, and pranoprofen). However, long-term use of corticosteroids may cause serious side effects such as increased intraocular pressure (steroid-induced glaucoma), cataracts, and secondary infections; NSAIDs have limited anti-inflammatory efficacy and may cause corneal epithelial damage. Therefore, the development of novel treatment strategies with novel mechanisms of action and high safety is urgently needed.

[0003] Recent studies have revealed that ferroptosis—an iron-dependent programmed cell death mechanism characterized by the accumulation of lipid peroxides—plays a crucial role in the occurrence and development of ocular surface inflammation. Various stressors (such as hyperosmolarity, oxidative stress, and stimulation by inflammatory factors) induce ferroptosis in corneal epithelial cells, leading to epithelial barrier disruption. The "damage" signals released by cells further recruit inflammatory cells such as neutrophils, exacerbating the local inflammatory cascade and forming a vicious cycle of "cell damage-inflammation." Therefore, intervening in this initial stage—inhibiting ferroptosis—potentially offers a promising new therapeutic approach by blocking the inflammatory process at its source.

[0004] Ferroplasmosis inhibitors, such as Ferrostatin-1 (Fer-1), can effectively scavenge lipid free radicals and inhibit the lipid peroxidation chain reaction, demonstrating strong ocular surface protection potential in preclinical models. However, small molecule inhibitors such as Fer-1 face significant drug development bottlenecks: their poor water solubility makes it difficult to formulate highly effective ocular preparations, severely limiting their clinical translation and application.

[0005] Cyclodextrin inclusion technology is a classic strategy for improving the solubility of hydrophobic drugs, but its effect on improving ocular surface retention and permeation is limited. Liposomes, as a mature nanodelivery carrier, possess good biocompatibility, bioadhesion, and sustained-release properties, effectively prolonging the residence time of drugs on the corneal surface and potentially promoting drug penetration through fusion with biological membranes. However, simply loading hydrophobic Fer-1 into the liposomal lipid bilayer presents problems such as low drug loading capacity and easy drug leakage.

[0006] Therefore, there is an urgent need for an innovative delivery strategy that can synergistically address multiple challenges such as the solubility and stability of ferroptosis inhibitors. Summary of the Invention

[0007] This invention aims to overcome the shortcomings of existing technologies and provide an ocular delivery system for ferroptosis inhibitors based on cyclodextrin inclusion and liposome loading, along with its preparation method and applications. This system is highly efficient, stable, and exhibits good ocular surface adaptability. The delivery system combines the solubilizing and stabilizing properties of cyclodextrin with the bioadhesive, sustained-release, and permeation-enhancing advantages of liposomes. It effectively prolongs the drug's residence time on the ocular surface, promotes transcorneal absorption, and targets inflammatory lesions on the ocular surface, efficiently inhibiting ferroptosis and thus achieving significant therapeutic effects on ocular surface inflammatory diseases such as dry eye and keratoconjunctivitis.

[0008] To achieve the above objectives, the present invention adopts the following technical solution:

[0009] An ocular delivery system for treating ocular surface inflammatory diseases based on cyclodextrin inclusion and liposome loading of a ferroptosis inhibitor, the delivery system comprising: (a) An inclusion complex formed by ferroptosis inhibitor and cyclodextrin; (b) The inclusion complex was loaded into the internal aqueous phase of the liposome vesicle.

[0010] The core of this invention lies in constructing a composite nanostructure of "cyclodextrin inclusion complex-liposome".

[0011] Specifically, this invention first utilizes the host-guest interaction between hydrophilic cyclodextrin and ferroptosis inhibitors in an aqueous phase to form a soluble inclusion complex. This step fundamentally transforms water-insoluble drug molecules into a soluble state, forming a water-soluble inclusion complex, significantly improving its solubility, and potentially enhancing the chemical stability of the drug through inclusion. Subsequently, the aqueous solution containing the drug (ferroptosis inhibitor) is used as the inner aqueous phase, and during the preparation of liposomes, it is encapsulated within the internal aqueous chambers (vesicles) of liposomes composed of phospholipids and cholesterol, forming an "inclusion complex-liposome" dual drug delivery system. In this structure, the phospholipid bilayer of the liposome acts as an external barrier and protective layer, providing sustained release, leakage prevention, and bioadhesion functions; while the encapsulated cyclodextrin-drug inclusion complex ensures that the drug exists in a dissolved state, achieving high drug loading and stability.

[0012] Preferably, the ferroptosis inhibitor includes at least one of Ferrostatin-1 (Fer-1), SRS16-86, UAMC-3203, and Liproxstatin-1.

[0013] Preferably, the cyclodextrin is selected from β-cyclodextrin (β-CD), hydroxypropyl-β-cyclodextrin (HP-β-CD), and hydroxyethyl-β-cyclodextrin. β At least one of the following: cyclodextrin (HE-β-CD), sulfobutyl-β-cyclodextrin (SBE-β-CD), methyl-β-cyclodextrin (M-β-CD), and glucose-β-cyclodextrin (Glucose-β-CD).

[0014] Preferably, the preparation of the cyclodextrin inclusion complex is carried out in a buffer solution with pH 4.0 to 7.5 (such as phosphate buffer, borate buffer, Tris-HCl buffer, etc.), preferably pH 6.5 to 7.5.

[0015] Preferably, a stabilizer is added during the preparation of the cyclodextrin inclusion complex. The stabilizer is selected from at least one of polyvinyl alcohol (PVA), disodium ethylenediaminetetraacetate (EDTA-2Na), poloxamer 188, and Tween 80, and its addition amount is 0.01 to 0.5% of the total weight of the system, preferably 0.1 to 0.3%.

[0016] Preferably, the preparation temperature of the cyclodextrin inclusion complex is 25-50 °C.

[0017] Preferably, the preparation of the cyclodextrin inclusion complex is carried out under stirring at a speed of 150 to 500 r / min.

[0018] Preferably, the inclusion time of the cyclodextrin inclusion complex is 2 to 6 hours.

[0019] Preferably, the molar ratio of the cyclodextrin to the ferroptosis inhibitor is 1:1 to 10:1, and more preferably 1:1 to 5:1.

[0020] Preferably, the liposomes are composed of phospholipids and cholesterol.

[0021] Preferably, the phospholipid is selected from at least one of hydrogenated soybean phospholipid, egg yolk lecithin, dipalmitoylphosphatidylcholine, and distearate phosphatidylcholine.

[0022] Preferably, the mass ratio of phospholipid to cholesterol is 1:1 to 10:1, and more preferably 1:1 to 5:1.

[0023] Preferably, the particle size range of the inclusion complex is 1–30 nm, and more preferably 5–15 nm.

[0024] The method for preparing the above delivery system includes the following steps: (a) The ferroptosis inhibitor was dissolved in an organic solvent to obtain a ferroptosis inhibitor solution; cyclodextrin was dissolved in a buffer solution to obtain a cyclodextrin solution; the ferroptosis inhibitor solution and the cyclodextrin solution were mixed and reacted to prepare a cyclodextrin-ferroptosis inhibitor inclusion complex solution. (b) Phospholipids and cholesterol are dissolved in an organic solvent and then rotary evaporated to form a lipid film; (c) Using the inclusion complex solution obtained in step (a) as the inner aqueous phase, hydrate the lipid film obtained in step (b) to prepare liposomes loaded with inclusion complex; (d) The liposomes obtained in step (c) are subjected to ultrasonic treatment to finally obtain nanoliposomes with a particle size range of 50 to 300 nm, preferably 50 to 150 nm.

[0025] Preferably, in step (a), the organic solvent is selected from ethanol and dimethyl sulfoxide.

[0026] Preferably, in step (a), the concentration of the ferroptosis inhibitor solution is 1 to 10 mg / mL.

[0027] Preferably, in step (a), the concentration of the cyclodextrin solution is 5 to 10 mg / mL.

[0028] Preferably, in step (a), the reaction is carried out under stirring at a speed of 150 to 500 r / min, preferably 300 r / min.

[0029] Preferably, in step (a), the reaction is carried out at 25-50°C for 2-6 h, then cooled to 10-25°C and stirred for 1-4 h, preferably at 40°C for 2 h, then cooled to room temperature and stirred for 2 h.

[0030] Preferably, in step (a), when a stabilizer is used, the stabilizer is dissolved in a buffer solution.

[0031] Preferably, in step (b), the organic solvent is selected from one or more of methanol, ethanol, chloroform, and dichloromethane, preferably a single solvent of methanol, ethanol, chloroform, and dichloromethane, or a mixed solvent of methanol and chloroform, or ethanol and chloroform, wherein the volume ratio of methanol to chloroform in the mixed solvent of methanol and chloroform is 1:1 to 4:1, and the volume ratio of ethanol to chloroform in the mixed solvent of ethanol and chloroform is 1:1 to 4:1.

[0032] Preferably, in step (b), the concentration of cholesterol in the organic solvent is 0.5 to 5 mg / mL.

[0033] Preferably, in step (b), the temperature of the rotary evaporation is 30 to 60°C, and more preferably 40°C.

[0034] Preferably, in step (c), the hydration conditions are: temperature 20-50°C and time 30-60 min, preferably 50°C and 30 min.

[0035] Preferably, in step (d), the conditions for ultrasonic treatment are: power 100-300 W, time 3-10 min, preferably ultrasonic for 2-10 s, pause for 2-10 s, and more preferably: power 150 W, time 5 min, preferably ultrasonic for 5 s, pause for 10 s.

[0036] Preferably, in step (d), the ultrasonic treatment is followed by filtration through a filter membrane to obtain nanoliposomes; wherein the pore size of the filter membrane is 0.1 to 0.5 μm, preferably 0.22 μm.

[0037] Preferably, the particle size of the nanoliposomes is in the range of 50 to 300 nm, and more preferably 50 to 150 nm.

[0038] A pharmaceutical composition comprising a delivery system as described above and a pharmaceutically acceptable ophthalmic excipient, said ophthalmic excipient comprising at least one of an isotonic adjuster (such as sodium chloride, boric acid, glycerin, etc.), a pH adjuster (such as a phosphate buffer system, a borate buffer system, an acetate buffer system, etc.), a thickener (such as hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, hyaluronic acid, polyvinyl alcohol, carbomer, etc.), and a preservative (such as benzalkonium chloride, chlorobutanol, ethylparaben, etc.).

[0039] The use of the above-described delivery system or pharmaceutical composition in the preparation of medicaments for the prevention and / or treatment of ocular surface inflammatory diseases. The ocular surface inflammatory diseases are selected from dry eye syndrome and related inflammation, keratoconjunctivitis, ocular surface damage caused by chemical burns, autoimmune ocular surface diseases, or ocular surface inflammatory diseases related to ferroptosis (dry eye syndrome, corneal damage, etc.).

[0040] The delivery system of the present invention can be easily formulated into any dosage form suitable for topical administration of ocular surface (such as eye drops, ointments, ophthalmic gels, etc.), and is suitable for long-term, frequent ocular administration, providing a novel, mechanism-driven treatment option for the clinical treatment of refractory ocular surface inflammation.

[0041] Compared with the prior art, the present invention has the following significant advantages: (1) Dual solubilization and protection: Cyclodextrin inclusion solves the hydrophobicity problem at the drug molecule level, while liposomes provide a nanoscale dispersion and protective environment; (2) Synergistic permeation and sustained release: The bioadhesiveness of liposomes prolongs the residence time of the formulation on the ocular surface, and its interaction with the corneal epithelium may promote drug permeation, while the presence of inclusion complexes ensures the stable storage and sustained release of the drug inside the liposomes; (3) Improve treatment efficiency and safety: By improving local bioavailability, potential systemic side effects can be reduced; (4) Wide range of applications: Based on the core mechanism of ferroptosis inhibition, it has potential therapeutic effects on ocular surface inflammation caused by a variety of etiologies (immunity, infection, physical and chemical damage). Attached Figure Description

[0042] Figure 1 Particle size distribution of Fer-1 / SBE-β-CD inclusion complex; Figure 2 Particle size distribution of Lip@Fer-1 / SBE-β-CD liposomes; Figure 3 Stability evaluation of Lip@Fer-1 / SBE-β-CD liposomes: (A) particle size, PDI value and (B) Zeta potential of liposomes within 28 days; Figure 4Safety evaluation of Lip@Fer-1 / SBE-β-CD liposome application; Figure 5 Evaluation of the antiferroptosis and anti-inflammatory activity of Lip@Fer-1 / SBE-β-CD liposomes at the cellular level; (A) Cell viability assay to evaluate the effect of Lip@Fer-1 / SBE-β-CD liposomes in rescuing ferroptosis; (B) Detection of intracellular MDA content in cells treated with Lip@Fer-1 / SBE-β-CD liposomes; (C) Secretion of cellular inflammatory factors IL-6 and TNF-α. Detailed Implementation

[0043] The following specific embodiments are for further explanation of the content of the present invention and should not be construed as limiting the present invention in any way.

[0044] Example 1 Preparation of Ferrostatin-1 / Sulfobutyl-β-cyclodextrin (Fer-1 / SBE-β-CD) inclusion complex: Fer-1 (13.1 mg) and SBE-β-CD (molar ratio 1:2, approximately 130 mg) were weighed. Fer-1 was dissolved in 5 mL of ethanol to obtain a Fer-1 ethanol solution, and SBE-β-CD was dissolved in 20 mL of phosphate buffer solution (pH 7.4) containing 0.2% Tween 80 to obtain an SBE-β-CD aqueous solution. The temperature of the SBE-β-CD aqueous solution was set to 40°C and the stirring speed was set to 300 r / min. The Fer-1 ethanol solution was slowly (dropwise) added to the SBE-β-CD aqueous solution, and the mixture was stirred for 2 h. The mixture was then cooled to room temperature and stirred for another 2 h to obtain a Fer-1 / SBE-β-CD inclusion complex solution. The particle size of the Fer-1 / SBE-β-CD inclusion complex was determined to be 11.2 ± 1.5 nm by dynamic light scattering. Figure 1 As shown.

[0045] Example 2 Preparation of liposomes loaded with Fer-1 / SBE-β-CD inclusion complex (Lip@Fer-1 / SBE-β-CD): Hydrogenated soybean lecithin (HSPC, 60 mg) (purchased from Beijing Innocare Technology Co., Ltd.) and cholesterol (Cholesterol, 20 mg) (purchased from Beijing Innocare Technology Co., Ltd.) were weighed and dissolved in 10 mL of ethanol / chloroform (2:1, v / v) mixed solvent to obtain a lipid solution. This solution was placed in a round-bottom flask and rotary evaporated in a 40°C water bath until a uniform thin film formed on the flask wall. The solution was then vacuum dried (60°C, 2 h) to completely remove the organic solvent. The Fer-1 / SBE-β-CD inclusion complex solution obtained in Example 1 was preheated to 50°C, and 10 mL was added to the flask containing the lipid film. The solution was rotary hydrated in a 50°C water bath for 30 min to obtain a liposome suspension. The liposome suspension was sonicated (150 W, 5 s sonication, 10 s pause, total duration 5 min), and finally filtered through a 0.22 μm sterile filter membrane to obtain Lip@Fer-1 / SBE-β-CD liposomes.

[0046] The average particle size of the liposomes obtained in Example 2 was determined to be 118.3 ± 5.2 nm. Figure 2 As shown, the polydispersity index (PDI) was 0.08 ± 0.01, the zeta potential was -8.5 ± 1.2 mV, and the encapsulation efficiency of Fer-1 was 85.7 ± 3.5%.

[0047] Example 3 Stability evaluation of Lip@Fer-1 / SBE-β-CD liposomes: On days 1, 3, 7, 14, 21 and 28 after the preparation of Lip@Fer-1 / SBE-β-CD liposomes in Example 2, the particle size, polydispersity index (PDI) and zeta potential of the samples were measured to examine the long-term stability of the liposomes.

[0048] The results are as follows Figure 3 As shown, the particle size, potential, and PDI remained stable over 28 days without significant changes, indicating that the prepared liposomes Lip@Fer-1 / SBE-β-CD have good long-term stability.

[0049] Example 4 Biocompatibility evaluation of Lip@Fer-1 / SBE-β-CD application: The safety of Lip@Fer-1 / SBE-β-CD was evaluated using the human corneal epithelial cell line (HCE-T). Experimental groups were set up as follows: (1) Background group: Contains no cells, only culture medium is added, used for background subtraction; (2) Blank control group: The cells were added to the culture medium and were not treated in any way. The cell viability was considered to be 100%. (3) Free drug group (Ferrostatin-1): Cells were incubated in a medium containing 5 μM Ferrostatin-1; (4) Liposome group (Lip@Fer-1 / SBE-β-CD): Cells were incubated in a culture medium containing Lip@Fer-1 / SBE-β-CD liposomes (5 μM Ferrostatin-1) prepared in Example 2.

[0050] 100 μL of each sample was added to a 96-well plate, with four replicates per group. The plates were placed in a cell culture incubator and incubated at 37°C and 5% CO2 for 24 h. Finally, 10 μL of CCK-8 working solution (purchased from Jiangsu Kaiji Biotechnology Co., Ltd.) was added to each well under dark conditions, and the 96-well plates were incubated at 37°C and 5% CO2 for 2 h. The absorbance (Optical Density, OD) of each well was measured at 450 nm using a microplate reader, and cell viability was calculated.

[0051] Figure 4 The results showed that the cell survival rate of the liposome group (Lip@Fer-1 / SBE-β-CD) was over 90%, which was not significantly different from the control group, indicating that the constructed Lip@Fer-1 / SBE-β-CD drug delivery system has good biocompatibility.

[0052] Example 5 Evaluation of ferroptosis and anti-inflammatory activity at the cellular level: The experimental groups are as follows: (1) Control group: Cells were cultured normally without any treatment; (2) Erastin model group: Cells were induced by the ferroptosis inducer Erastin (10 μM) to establish a ferroptosis cell model; (3) Ferrostatin-1 treatment group: First, ferroptosis model was induced by adding Erastin (10 μM) as an inducer of ferroptosis, and then the free drug Ferrostatin-1 (5 μM) was used alone for treatment; (4) Lip@Fer-1 / SBE-β-CD treatment group: First, ferroptosis model was induced by adding the ferroptosis inducer Erastin (10 μM), and then Lip@Fer-1 / SBE-β-CD liposomes (5 μM Ferrostatin-1) prepared in Example 2 were used for treatment.

[0053] Cells in the logarithmic growth phase were selected and subjected to a 1×10⁻⁶ ionization assay. 4Cells were seeded at a density of [number] cells / well in 96-well plates and cultured overnight at 37°C with 5% CO2. After complete cell adhesion, HCE-T cell ferroptosis models were constructed by inducing cell death in all groups except the control group with Erastin (10 μM) for 24 h. Subsequently, prepared cell culture medium containing drugs or drug-loaded liposomes was added to each group according to their respective groups. The control and model groups received only an equal volume (100 μL / well) of cell culture medium, and incubation continued for 24 h. After incubation, the plates were removed, and cell viability was detected by the CCK-8 assay, intracellular malondialdehyde content was detected by the thiobarbituric acid reaction assay, and the secretion levels of typical inflammatory factors IL-6 and TNF-α were detected by ELISA.

[0054] Figure 5 The results showed that, compared with the model group, both the free drug treatment group (Ferrostatin-1) (p<0.01) and the liposome treatment group (Lip@Fer-1 / SBE-β-CD) (p<0.001) significantly improved cell viability. Furthermore, at the same Fer-1 concentration, the Lip@Fer-1 / SBE-β-CD group showed significantly stronger protective effects than the Ferrostatin-1 group (p<0.05). Further analysis using malondialdehyde (MDA), a lipid peroxidation product, and ELISA to detect the expression of inflammatory factors IL-6 and TNF-α confirmed that Lip@Fer-1 / SBE-β-CD more effectively inhibited ferroptosis-related oxidative damage and downstream inflammatory responses.

Claims

1. An ocular delivery system of ferroptosis inhibitor based on cyclodextrin inclusion and liposome drug loading, characterized in that, The delivery system includes: (a) An inclusion complex formed by ferroptosis inhibitor and cyclodextrin; (b) The inclusion complex was loaded into the internal aqueous phase of the liposome vesicle.

2. The drug delivery system of claim 1, wherein, The ferroptosis inhibitor is selected from at least one of Ferrostatin-1, SRS16-86, UAMC-3203, and Liproxstatin-1; The cyclodextrin is selected from the group consisting of β-cyclodextrin, hydroxypropyl-β-cyclodextrin, hydroxyethyl β cyclodextrin, sulfobutyl-β-cyclodextrin, methyl-β-cyclodextrin, glucosyl-β-cyclodextrin; and the molar ratio of the cyclodextrin to the ferroptosis inhibitor is 1:1 ~ 10:

1.

3. The delivery system according to claim 1, characterized in that, The preparation of the cyclodextrin inclusion complex was carried out in a buffer solution with a pH of 4.0 to 7.

5. A stabilizer was also added during the preparation process. The stabilizer was selected from at least one of polyvinyl alcohol, disodium ethylenediaminetetraacetate, poloxamer 188, and Tween 80. The amount of stabilizer added was 0.01 to 0.5% of the total weight of the system.

4. The delivery system according to claim 1, characterized in that, The preparation conditions for the cyclodextrin inclusion complex are: temperature 25 ~ 50 ℃, stirring speed 150 ~ 500 r / min, and inclusion time 2 ~ 6 h.

5. The delivery system according to claim 1, characterized in that, The liposomes are composed of phospholipids and cholesterol; the phospholipids are selected from at least one of hydrogenated soybean phospholipids, egg yolk lecithin, dipalmitoylphosphatidylcholine, and distearate phosphatidylcholine; the mass ratio of the phospholipids to cholesterol is 1:1 to 10:

1.

6. A method for preparing a drug delivery system as described in any one of claims 1 to 5, characterized in that, The preparation steps include the following: (a) The ferroptosis inhibitor was dissolved in an organic solvent to obtain a ferroptosis inhibitor solution; cyclodextrin was dissolved in a buffer solution to obtain a cyclodextrin solution; the ferroptosis inhibitor solution and the cyclodextrin solution were mixed and reacted to prepare an inclusion complex solution; (b) Phospholipids and cholesterol are dissolved in an organic solvent and then rotary evaporated to form a lipid film; (c) Using the inclusion complex solution obtained in step (a) as the inner aqueous phase, hydrate the lipid film obtained in step (b) to prepare liposomes loaded with inclusion complex; (d) The liposomes obtained in step (c) are subjected to ultrasonic treatment to obtain nanoliposomes.

7. The preparation method according to claim 6, characterized in that, In step (a), the organic solvent is selected from ethanol and dimethyl sulfoxide, the concentration of the ferroptosis inhibitor solution is 1 to 10 mg / mL, and the concentration of the cyclodextrin solution is 5 to 10 mg / mL; when a stabilizer is used, the stabilizer is dissolved in a buffer solution; In step (b), the organic solvent is selected from one or more of methanol, ethanol, chloroform, and dichloromethane, and the concentration of cholesterol in the organic solvent is 0.5–5 mg / mL; the rotary evaporation temperature is 30–60°C. In step (c), the hydration conditions are: temperature 20–50°C and time 30–60 min; In step (d), the conditions for ultrasonic treatment are: power 100-300 W and time 3-10 min. After ultrasonic treatment, the ultrasonic treatment is further filtered through a filter membrane with a pore size of 0.22 μm.

8. A pharmaceutical composition, characterized in that, The ophthalmic excipient comprises a delivery system as described in any one of claims 1 to 5 and a pharmaceutically acceptable ophthalmic excipient, preferably comprising at least one of an isotonic adjuster, a pH adjuster, a thickener, and a preservative.

9. The use of the delivery system according to any one of claims 1 to 5 or the pharmaceutical composition according to claim 8 in the preparation of a medicament for the prevention and / or treatment of ocular surface inflammatory diseases.

10. The application according to claim 9, characterized in that, The ocular surface inflammatory diseases are selected from dry eye syndrome and related inflammation, keratoconjunctivitis, ocular surface damage caused by chemical burns, autoimmune ocular surface diseases, or ocular surface inflammatory diseases related to ferroptosis.

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