Application of PDL1 antibody in corneal neovascularization disease

Abstract

The present invention relates to the use of anti-PDL1 antibodies in the treatment of corneal neovascular diseases. The present invention constructs an animal model of corneal neovascularization induced by alkali burns, and performs slit lamp examination to evaluate corneal neovascularization and corneal vascular tissue immunofluorescence staining after topical application of anti-PDL1 antibodies. It is found that the corneal neovascularization of mice in the anti-PDL1 antibody treatment group is significantly reduced, indicating that topical application of anti-PDL1 antibodies can inhibit corneal neovascularization. It is confirmed that anti-PDL1 antibodies can significantly reduce PDL1 hi CXCR2 low The infiltration of neutrophil subsets inhibits the secretion of a series of inflammatory factors and vascular growth factors, thereby inhibiting pathological neovascularization. The present invention provides experimental basis and theoretical basis for the treatment of corneal neovascular diseases with anti-PDL1 antibodies and the expansion of clinical applications of anti-PDL1 antibodies, revealing that anti-PDL1 antibodies have clear clinical application prospects in the field of neovascular diseases.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the use of anti-PDL1 antibodies in the preparation of drugs for treating corneal neovascularization diseases. Background Art

[0002] Corneal neovascularization (CNV) is a common pathological feature of many ocular surface diseases, and its formation is an important cause of blindness caused by corneal diseases. Under normal physiological conditions, the cornea is avascular tissue and remains transparent. However, in cases of infection, trauma, inflammation, autoimmune diseases, corneal transplant rejection, etc., the blood vessels at the corneal limbus sprout and proliferate, forming pathological corneal neovascularization. At present, the main clinical treatments for corneal neovascularization are steroid hormones, nonsteroidal, immunosuppressive and anti-VEGF drugs. In severe cases, ocular surface reconstruction surgery is required. However, these treatments have limited efficacy and have adverse side effects such as increased intraocular pressure, cataracts, and glaucoma, and are not suitable for long-term use. Therefore, it is necessary to further explore the mechanism of corneal neovascularization and develop anti-neovascular therapy with clear targets for the pro-inflammatory and pro-neovascular cell types in the local pathological microenvironment.

[0003] Programmed death ligand 1 (PDL1) is overexpressed in a variety of malignant tumor cells. It binds to programmed death receptor 1 (PD1), a common immunosuppressive member on the surface of T cells, and plays a vital role in downregulating immune system activity and improving immune tolerance. Anti-PDL1 / PD1 therapy is a major breakthrough in the field of tumor immunotherapy research in recent years. Many anti-PDL1 / PD1 drugs have been approved by the FDA for anti-tumor treatment. With the in-depth study of the PD1 / PDL1 pathway, it has been found that it is also involved in a variety of autoimmune diseases (systemic lupus erythematosus, type 1 diabetes), Alzheimer's disease, pulmonary fibrosis, etc. In addition, some studies have shown that anti-PD1 / PDL1 is related to angiogenesis. In a mouse lung cancer model, anti-PDL1 antibodies can upregulate angiogenesis inhibitors CXCL10 / 11 and inhibit angiogenesis in lung cancer. In a mouse hindlimb ischemia model, inhibition of PD1 can upregulate IFN-γ to impair the proliferation and migration of vascular endothelial cells. However, in the field of anti-inflammatory neovascularization, especially in the treatment of corneal neovascular diseases, no studies have reported that anti-PDL1 antibodies inhibit corneal neovascularization. Summary of the invention

[0004] The purpose of the present invention is to solve the problems existing in the prior art in the treatment of corneal neovascularization diseases, thereby providing the use of anti-PDL1 antibodies in the preparation of drugs for treating corneal neovascularization diseases. Anti-PDL1 antibodies are used as active ingredients to effectively inhibit the growth of PDL1. hi CXCR2 lowThe pro-angiogenic neutrophil subpopulation with PDL1 as a marker infiltrates the cornea, reducing its secretion of pro-inflammatory factors and vascular growth factors, thereby significantly inhibiting local inflammation and pathological neovascularization. The application of anti-PDL1 antibodies can reduce corneal inflammation, has anti-inflammatory and anti-angiogenic effects, and provides a practical theoretical basis for the clinical treatment of corneal neovascular diseases.

[0005] In order to solve the above technical problems, the present invention is implemented through the following technical solutions.

[0006] The first aspect of the present invention provides the use of an anti-PDL1 antibody in the preparation of a drug for treating corneal neovascular diseases and / or improving physiological indicators related to corneal neovascular diseases.

[0007] Preferably, the physiological index related to corneal neovascular disease is selected from one or more of the area of ​​corneal neovascularization, the score of corneal neovascularization, the number of infiltrating corneal immune cells, the type of infiltrating corneal immune cells, and the secretion level of inflammatory neovascularization factors. Specifically, the anti-PDL1 antibody can reduce the area of ​​corneal neovascularization, reduce the score of corneal neovascularization, reduce the number of infiltrating corneal immune cells, and reduce the secretion level of inflammatory neovascularization factors.

[0008] Preferably, the infiltrating corneal immune cells are selected from neutrophils; most preferably, the infiltrating corneal immune cells are selected from PDL1 hi CXCR2 low Neutrophil markers.

[0009] Preferably, the inflammatory angiogenic factor is selected from one or more of Csf3, Il1a, Il1b, Il12a, Il23a, Tgfb1, Cxcl1, Cxcl2, Cxcl3, Ccl3, Ccl4, Mmp3, and Vegfa;.

[0010] A second aspect of the present invention provides a pharmaceutical composition for treating corneal neovascular diseases and / or improving physiological indicators related to corneal neovascular diseases, comprising an anti-PDL1 antibody and a phosphate buffer, wherein the concentration of the anti-PDL1 antibody is 6-8 mg / mL.

[0011] Preferably, the physiological index related to corneal neovascular disease is selected from one or more of the area of ​​corneal neovascularization, the score of corneal neovascularization, the number of infiltrating corneal immune cells, the type of infiltrating corneal immune cells, and the secretion level of inflammatory neovascularization factors. Specifically, the anti-PDL1 antibody can reduce the area of ​​corneal neovascularization, reduce the score of corneal neovascularization, reduce the number of infiltrating corneal immune cells, and reduce the secretion level of inflammatory neovascularization factors.

[0012] Preferably, the infiltrating corneal immune cells are selected from neutrophils; most preferably, the infiltrating corneal immune cells are selected from PDL1 hi CXCR2 low Neutrophil markers.

[0013] Preferably, the inflammatory angiogenic factor is selected from one or more of Csf3, Il1a, Il1b, Il12a, Il23a, Tgfb1, Cxcl1, Cxcl2, Cxcl3, Ccl3, Ccl4, Mmp3, and Vegfa.

[0014] The third aspect of the present invention provides a pharmaceutical preparation for treating corneal neovascular diseases and / or improving physiological indicators related to corneal neovascular diseases, comprising an anti-PDL1 antibody, a phosphate buffer and a pharmaceutically acceptable excipient, wherein the concentration of the anti-PDL1 antibody is 6-8 mg / mL.

[0015] Preferably, the physiological index related to corneal neovascular disease is selected from one or more of the area of ​​corneal neovascularization, the score of corneal neovascularization, the number of infiltrating corneal immune cells, the type of infiltrating corneal immune cells, and the secretion level of inflammatory neovascularization factors. Specifically, the anti-PDL1 antibody can reduce the area of ​​corneal neovascularization, reduce the score of corneal neovascularization, reduce the number of infiltrating corneal immune cells, and reduce the secretion level of inflammatory neovascularization factors.

[0016] Preferably, the infiltrating corneal immune cells are selected from neutrophils; most preferably, the infiltrating corneal immune cells are selected from PDL1 hi CXCR2 low Neutrophil markers.

[0017] Preferably, the inflammatory angiogenic factor is selected from one or more of Csf3, Il1a, Il1b, Il12a, Il23a, Tgfb1, Cxcl1, Cxcl2, Cxcl3, Ccl3, Ccl4, Mmp3, and Vegfa.

[0018] Preferably, the dosage form of the pharmaceutical preparation is selected from one or more of a solution, a sol, an emulsion, and a suspension.

[0019] Preferably, the dosage form of the pharmaceutical preparation is selected from one or more of a solution, a sol, an emulsion, and a suspension.

[0020] Preferably, the dosage form of the pharmaceutical preparation is selected from a solution.

[0021] Preferably, the pharmaceutically acceptable excipient is selected from one or more of a solvent, a solubilizer, a surfactant, an antibacterial agent, an antioxidant, and a chelating agent.

[0022] It should be noted that, unless otherwise specified, the "anti-PDL1 antibody" or similar expressions in the context of the present invention refer to antibody types that can specifically interact with programmed death ligand 1 (PDL1), including but not limited to monoclonal antibodies, polyclonal antibodies, etc., which can be obtained commercially or prepared by themselves. In order to fully display the relevant experiments and technical solutions, and to facilitate the understanding of those skilled in the art, the commercially available anti-PDL1 antibody (BioXCell InVivoPlus anti-mouse PD-L1 (B7-H1), item number BP0101) is used in the context of the present invention, but the antibody used is not used to limit the technical solution and protection scope of the present invention, and all antibodies that can specifically interact with PDL1 should be understood as the scope of the "anti-PDL1 antibody" mentioned in the present invention.

[0023] Compared with the prior art, the present invention has the following technical effects:

[0024] (1) Through a large number of studies, the present invention has fully confirmed the therapeutic effect of anti-PDL1 on corneal neovascularization. Among them, anti-PDL1 antibodies mainly clear PDL1 infiltrates after corneal damage. hi CXCR2 low Neutrophil subsets inhibit the levels of local Vegfa, Il-1b and other factors, improve the corneal inflammatory microenvironment, and reduce the area of ​​pathological neovascularization. Anti-PDL1 antibodies have the effect of inhibiting pathological neovascularization, improving corneal function, and opening up new treatment ideas for the clinical treatment of corneal neovascularization diseases.

[0025] (2) The present invention conducts in-depth research on the mechanism of anti-PDL1 antibodies in the treatment of corneal neovascularization diseases and clarifies that anti-PDL1 antibodies can inhibit corneal PDL1 hi CXCR2 low The number of neutrophil subpopulation infiltrations can be reduced, and the release of pro-inflammatory factors and angiogenic factors can be inhibited. The application of anti-PDL1 antibodies can improve the corneal inflammatory microenvironment caused by these cells, providing a practical theoretical basis for the subsequent treatment and drug development of corneal neovascular diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Schematic diagram of the slit lamp and the statistical graph of the neovascularization score for the therapeutic effect of anti-PDL1 antibody on corneal neovascularization after alkali burn in mice.

[0027] Figure 2This is a statistical diagram of corneal neovascularization and neovascularization area after CD31 staining after treatment of corneal neovascularization in mice with alkali burns using anti-PDL1 antibody.

[0028] Figure 3 After anti-PDL1 treatment, the mouse cornea was infiltrated with PDL1 hi CXCR2 low Representative diagram of neutrophil subset infiltration.

[0029] Figure 4 After anti-PDL1 treatment, the mouse cornea expressed inflammatory factors (Il1b) and angiogenic factors (Vegfa) levels.

[0030] Figure 5 Single-cell transcriptome sequencing of the cornea after alkali burn showed the mRNA expression levels of Cd274 (PDL1), Cxcr2, Csf3, Il1a, Il1b, Il12a, Il23a, Tgfb1, Cxcl1, Cxcl2, Cxcl3, Mmp3, and Vegfa in various subsets of infiltrating mouse neutrophils.

[0031] Figure 6 After subconjunctival injection of IgG and anti-PDL1 antibody, transcriptome sequencing of the alkali-burned cornea was performed to determine the mRNA expression levels of Csf3, Il1a, Il1b, Il12a, Il23a, Tgfb1, Cxcl1, Cxcl2, Cxcl3, Ccl3, Ccl4, Mmp3, and Vegfa in the mouse cornea. DETAILED DESCRIPTION

[0032] In order to make the purpose, technical scheme and effect of the present invention clearer and more specific, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0033] Unless otherwise specified, the reagents, consumables and instruments used in the present invention were all commercially available. Animal ethics and all experimental operations were in compliance with the Regulations on the Administration of Laboratory Animals of the State Science and Technology Commission. The experimental methods used in the present invention, such as immunohistochemistry, cell experiments, animal experiments, etc., are all conventional methods and techniques in the art.

[0034] Representative results from repeated biological experiments are presented in the context figures, and data are presented as mean ± SEM as specified in the figures. All in vitro experiments were repeated at least three times, and animal experiments were repeated twice. Data were analyzed using GraphPad Prism 9.0 or SPSS 22.0 software. Conventional medical statistical methods such as t-test, chi-square test, and analysis of variance were used to compare the mean differences between two or more groups.p A value < 0.05 was considered a significant difference.

[0035] Example 1 Construction of disease model

[0036] Alkali burns are a common type of chemical injury to the eye. Alkaline substances have strong penetration and destructiveness to tissues, which often cause complications such as corneal ulcers and perforations, and the formation of pathological neovascularization, which seriously affect vision. In animal models, the corneal neovascularization model induced by alkali burns is an inflammatory model with strong sensitivity, good reproducibility, and consistent response. It is simple to make and easy to observe the effects, providing an ideal platform for the mechanism and treatment of inflammatory corneal neovascularization.

[0037] The present invention uses C57BL / 6J-Irf8- / - mice to construct an alkali burn corneal neovascularization model. The specific steps are as follows:

[0038] (1) Cut a 2 mm diameter filter paper disc and soak it in 0.5 mol / L NaOH solution for later use;

[0039] (2) 8-12 week old C57BL / 6J-Irf8- / - mice were randomly divided into three groups, with 10 mice in each group;

[0040] (3) Group 1 (C57BL / 6J-Irf8- / - blank group) was a blank group without intervention; Group 2 (C57BL / 6J-Irf8- / - alkali burn-induced CNV model group + IgG), a filter paper soaked in alkali solution was attached to the front of the cornea for 30 seconds, then rinsed with a large amount of saline for 30 seconds to form a central disc-shaped burn area of ​​the cornea, and then IgG (BioXCell InVivoPlus rat IgG2bIsotype control, catalog number BP0090) 5uL was injected subconjunctivally; Group 3 (C57BL / 6J-Irf8- / - alkali burn-induced CNV model group + anti-PDL1 antibody), a filter paper soaked in alkali solution was attached to the front of the cornea for 30 seconds, then rinsed with a large amount of saline for 30 seconds to form a central disc-shaped burn area of ​​the cornea, and then 5uL of anti-PDL1 antibody was injected subconjunctivally;

[0041] (4) The modeling day was D0. IgG or anti-PDL1 antibody was repeatedly injected on D3 and D6, respectively. Samples were collected on D7 to evaluate the treatment effect.

[0042] Example 2 Slit lamp examination to evaluate corneal alkali burn neovascularization and scoring

[0043] (1) On the 7th day after the corneal alkali burn modeling in Example 1, ocular surface photography of the mice in each group was performed using a slit lamp (BX-900). The mice were anesthetized by intraperitoneal injection of 0.5% sodium pentobarbital (0.1 mL / 10 g);

[0044] (2) Fix the mouse and expose the cornea for photography under a 25X lens;

[0045] (3) The degree of corneal neovascularization in mice was scored as follows: no neovascularization: 0 points; neovascularization at the limbus: 1 point; neovascularization reaching the center of the cornea: 2 points; neovascularization coiled in the middle of the cornea: 3 points.

[0046] Test results such as Figure 1 The results showed that the cornea of ​​the blank group mice was transparent, with only vascular rings visible at the limbus; a large number of pathological new blood vessels were seen at the limbus in the IgG model group, invading the central cornea; and the area of ​​pathological new blood vessels in the PDL1 treatment group was significantly reduced. This shows that anti-PDL1 antibodies can effectively inhibit the formation of pathological new blood vessels in the cornea.

[0047] Example 3 Immunofluorescence staining of CD31 on corneal flat mounts to show the area of ​​new blood vessels

[0048] (1) On the 7th day after the alkali burn modeling in Example 1, the mice in each group were killed by cervical dislocation, and the eyeballs were obtained and fixed in 20 times the volume of 4% PFA for 60 minutes;

[0049] (2) Fix the eyeball with toothed forceps under a dissecting microscope, remove the optic nerve first, remove the connective tissue and muscles of the eyeball wall, cut the sclera, remove the iris, lens, retina and other tissues, and obtain a complete cornea;

[0050] (3) The cornea was blocked in 0.5% Triton X-100 / PBS + 5% BSA at room temperature for 1 hour or at 4°C overnight;

[0051] (4) Freshly prepare the primary antibody for immunofluorescence (CD31): dilute it in 5% BSA at a ratio of 1:100, place the cornea in the antibody, and incubate it at 4°C overnight. During this period, gently shake the EP tube every few hours to ensure full contact with the antibody;

[0052] (5) Remove the cornea and incubate with secondary antibody at room temperature for 1-2 hours, keeping away from light; wash with PBST 6 times, 10 minutes each time;

[0053] (6) Transfer the cornea to a glass slide and remove impurities with toothless forceps under a anatomical stereomicroscope. Make four radial cuts with the corneal epithelium facing upwards. Add a small amount of anti-fluorescence quenching sealing agent to seal the slide. Observe and photograph under a fluorescence microscope or confocal microscope.

[0054] The results are as follows Figure 2The results showed that the cornea of ​​the blank mice was transparent, with only vascular rings visible at the limbus; a large number of pathological new blood vessels were seen at the limbus of the IgG model group, invading the center of the cornea; the area of ​​pathological new blood vessels in the cornea of ​​the PDL1 treatment group was significantly reduced, *** p <0.001. The above results further prove that anti-PDL1 antibody can effectively reduce the formation of corneal pathological neovascularization.

[0055] Example 4 Immunofluorescence detection of infiltrated PDL1 in corneal frozen sections hi CXCR2 low Changes in the levels of immune cells and secreted cytokines

[0056] (1) On the 7th day after the alkali burn modeling in Example 1, the mice in each group were killed by cervical dislocation, and the eyeballs were obtained and fixed in 20 times the volume of 4% PFA for 60 minutes;

[0057] (2) Dehydrate in 10%-30% gradient sucrose, and after the tissue sinks to the bottom, embed it in OCT; slice 8-10 μm sections with a slicer and attach them to a slide;

[0058] (3) The sections were blocked in 0.5% Triton X-100 / PBS + 5% BSA at room temperature for 1 hour or at 4°C overnight;

[0059] (4) Freshly prepared primary antibodies (PDL1 / CXCR2 and Vegfa / Il1b) were diluted 1:100 in 5% BSA, and the sections were incubated in the antibodies at 4°C overnight.

[0060] (5) Remove the cornea and incubate with secondary antibody at room temperature for 1-2 hours, keeping away from light; wash with PBST 6 times, 10 minutes each time;

[0061] (6) Seal the slides with mounting medium and photograph them using a confocal microscope.

[0062] Test results such as Figure 3-4 The results showed that after the treatment group was treated with anti-PDL1 antibody, the infiltrated PDL1 hi CXCR2 low Cytoreduction,** p <0.01. The levels of local corneal Vegfa and Il1b factors decreased, indicating that the application of anti-PDL1 antibodies can reduce corneal infiltration of PDL1 hi CXCR2 low cells, reducing the corneal inflammatory environment.

[0063] Example 5 Effect of PDL1 Antibody on Secretion Levels of Inflammatory Angiogenic Factors in Mouse Cornea

[0064] (1) On the 7th day after the corneal alkali burn modeling in Example 1, the mice in each group were killed by cervical dislocation, the eyeballs were obtained, the corneas were separated, and digested in a mixed enzyme of collagenase and DNA at 37°C in a water bath for 45 minutes;

[0065] (2) Terminate digestion with 10% FBS, repeatedly blow until no clumps of tissue can be seen with the naked eye, and filter the cells with a 70 μm filter;

[0066] (3) When the cell viability is calculated to be ≥80%, the concentration of the cell suspension is adjusted to 106 / ml and the cells are prepared into a single-cell system using the 10X Genomics platform;

[0067] (4) Perform reverse transcription, emulsion breaking, cDNA library construction, and sequencing;

[0068] (5) The raw sequencing data were aligned to the mouse reference genome, exported through Cell Ranger, and analyzed using Seurat.

[0069] Test results such as Figure 5 The results showed that the neutrophils infiltrating the cornea after alkali burn were mainly divided into five subgroups, among which the N0 subgroup was PDL1 hi CXCR2 low In cell subsets, N0 upregulated the mRNA levels of inflammatory angiogenic factors Csf3, Il1a, Il1b, Il12a, Il23a, Tgfb1, Cxcl1, Cxcl2, Cxcl3, Mmp3, and Vegfa.

[0070] Furthermore, corneal transcriptome sequencing showed the secretion level of inflammatory angiogenic factors in the mouse cornea after subconjunctival injection of IgG and anti-PDL1 antibody. The specific steps are as follows:

[0071] (1) On D7 after the alkaline burn modeling and subconjunctival IgG and anti-PDL1 antibody injection in Example 1, the mice in each group were killed by cervical dislocation, the eyeballs were obtained, the corneas were separated, and every 4 corneas were directly placed in 1 ml of pre-cooled Trizol and repeatedly blown until the tissue was completely dissolved;

[0072] (2) After RNA was extracted using the Trizol method, agarose gel electrophoresis was used to analyze RNA integrity and the presence of DNA contamination;

[0073] (3) Total RNA was constructed using the Illumina NEBNext® Ultra™ RNA Library Prep Kit, followed by preliminary quantification using Qubit2.0 Fluorometer, library testing using Agilent 2100 bioanalyzer, and accurate quantification using q-RT RCR, before Illumina sequencing.

[0074] (4) The sequence data were indexed with a reference genome using HISAT2 v2.0.5, and paired-end clean reads were aligned to the reference genome using HISAT2 v2.0.5;

[0075] (5) Feature Counts is used to calculate the number of reads mapped to each gene, calculate the FPKM of each gene based on the length of the gene, and calculate the number of reads mapped to the gene.

[0076] Test results such as Figure 6 The results showed that after subconjunctival injection of anti-PDL1 antibody, the mRNA expression levels of Csf3, Il1a, Il1b, Il12a, Il23a, Tgfb1, Cxcl1, Cxcl2, Cxcl3, Ccl3, Ccl4, Mmp3, and Vegfa in the cornea of ​​alkali-burned mice decreased compared with those injected with IgG. This further indicates that the application of anti-PDL1 antibody can reduce the corneal infiltration of neutrophils, reduce the corneal inflammatory environment, and inhibit the formation of new blood vessels.

[0077] In summary, the present invention confirms through slit lamp ocular surface photography, corneal immunofluorescence staining and other experiments that subconjunctival injection of anti-PDL1 antibodies can reduce PDL1 hi CXCR2 low Cells infiltrate the cornea, significantly reduce the release of local inflammatory factors and angiogenic factors, improve the corneal inflammatory microenvironment, and reduce corneal neovascularization. The present invention provides experimental evidence and theoretical basis for the potential clinical application of anti-PDL1 antibodies in the treatment of corneal neovascular diseases and anti-PDL1 antibodies as neovascularization inhibitors. Anti-PDL1 antibodies have clear clinical application prospects in the preparation of pharmaceutical preparations for the treatment of corneal neovascular diseases.

[0078] The above specific implementation method part specifically introduces the analytical method involved in the present invention. It should be noted that the above introduction is only to help those skilled in the art better understand the method and ideas of the present invention, rather than limiting the relevant content. Without departing from the principle of the present invention, those skilled in the art may also make appropriate adjustments or modifications to the present invention, and the above adjustments and modifications shall also fall within the scope of protection of the present invention.

Claims

1. Use of an anti-PDL1 antibody in the preparation of a drug for treating corneal neovascular diseases and / or improving physiological indicators related to corneal neovascular diseases, characterized in that: The physiological indicators related to corneal neovascularization disease are selected from one or more of the area of ​​corneal neovascularization, the score of corneal neovascularization, the number of infiltrating corneal immune cells, the type of infiltrating corneal immune cells, and the secretion level of inflammatory neovascularization factors.

2. The use according to claim 1, characterized in that: The infiltrating corneal immune cells are selected from PDL1 hi CXCR2 low Neutrophil markers.

3. The use according to claim 1, characterized in that: The inflammatory angiogenesis-promoting factors are selected from one or more of Csf3, Il1a, Il1b, Il12a, Il23a, Tgfb1, Cxcl1, Cxcl2, Cxcl3, Ccl3, Ccl4, Mmp3, and Vegfa.

4. A pharmaceutical composition for treating corneal neovascularization and / or improving physiological indicators related to corneal neovascularization, characterized in that: It comprises anti-PDL1 antibody and phosphate buffer, wherein the concentration of the anti-PDL1 antibody is 6-8 mg / mL; the physiological indicators related to corneal neovascularization disease are selected from one or more of the area of ​​corneal neovascularization, the score of corneal neovascularization, the number of infiltrating corneal immune cells, the type of infiltrating corneal immune cells, and the secretion level of inflammatory pro-angiogenic factors.

5. The pharmaceutical composition according to claim 4, characterized in that The infiltrating corneal immune cells are selected from PDL1 hi CXCR2 low Neutrophil markers.

6. The pharmaceutical composition according to claim 4, characterized in that The inflammatory angiogenesis-promoting factors are selected from one or more of Csf3, Il1a, Il1b, Il12a, Il23a, Tgfb1, Cxcl1, Cxcl2, Cxcl3, Ccl3, Ccl4, Mmp3, and Vegfa.

7. A pharmaceutical preparation for treating corneal neovascularization and / or improving physiological indicators related to corneal neovascularization, characterized in that: The invention comprises an anti-PDL1 antibody, a phosphate buffer and a pharmaceutically acceptable excipient, wherein the concentration of the anti-PDL1 antibody is 6-8 mg / mL; and the physiological indicators related to corneal neovascularization disease are selected from one or more of the area of ​​corneal neovascularization, the score of corneal neovascularization, the number of infiltrating corneal immune cells, the type of infiltrating corneal immune cells, and the secretion level of inflammatory pro-angiogenic factors.

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