Application of anti-interferon-gamma neutralizing antibodies in the preparation of drugs for the prevention and / or treatment of dry eye syndrome
By inhibiting interferon-γ with anti-interferon-γ neutralizing antibodies and protecting lacrimal gland cells, this approach solves the treatment challenge of dry eye syndrome associated with Sjögren's syndrome, achieving corneal epithelial cell protection and tear secretion restoration, and providing a new targeted treatment strategy.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- BEIJING TONGREN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
- Filing Date
- 2026-04-03
- Publication Date
- 2026-06-30
AI Technical Summary
Existing treatments for dry eye have limitations, especially for dry eye associated with Sjögren's syndrome. The pathological mechanisms of existing drugs on the lacrimal glands are not clear, resulting in limited therapeutic effects. Furthermore, long-term use of artificial tears and corticosteroids has side effects.
By using an anti-interferon-γ neutralizing antibody and administering it via systemic injection, the release of interferon-γ is inhibited, lacrimal gland solid cells are protected, ferroptosis and immune cell infiltration are reduced, the JAK-STAT pathway is regulated, and a targeted therapy strategy is provided.
It significantly improves dry eye symptoms, reduces corneal defects, and restores tear secretion, providing a new and effective treatment for dry eye syndrome.
Smart Images
Figure CN122297665A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of biomedical technology, and in particular to the application of anti-interferon-γ neutralizing antibodies in the preparation of drugs for the prevention and / or treatment of dry eye syndrome. Background Technology
[0002] Sjögren's syndrome (SLS) is a systemic autoimmune disease characterized by pro-inflammatory lymphocytic infiltration in exocrine glands, primarily the lacrimal and salivary glands, leading to glandular dysfunction. The lacrimal glands secrete the lipid layer of the tear film, a fluid crucial for maintaining ocular surface health and optimal light refraction. In SLS, autoimmune attacks damage the lacrimal gland parenchyma, resulting in severe tear-deficiency dry eye; due to its complex pathophysiological mechanisms and high incidence, this disease has received increasing attention. Compared to non-Sjögren's syndrome-associated dry eye, SLS-associated dry eye often presents with more severe signs and a higher risk of vision-threatening complications, stemming from the specific damaging effects of its systemic autoimmune characteristics. Although the lacrimal glands are considered the primary target of Sjögren's syndrome, the exact mechanisms leading to dysfunction of its secretory parenchyma and subsequent functional failure are not fully elucidated, thus requiring a more detailed pathological atlas to describe its comprehensive pathological changes.
[0003] Apoptosis has long been considered the dominant mode of cell death in Sjögren's syndrome. However, recent studies have revealed ferroptosis—a unique form of regulated cell death triggered by iron-dependent lipid peroxidation—as a novel pathogenic factor in a range of autoimmune and inflammatory diseases. The mechanism of ferroptosis is driven by the accumulation of lipid peroxides, and the antioxidant enzyme glutathione peroxidase 4 is crucial in preventing its occurrence. While its role in other diseases is being explored, the involvement of ferroptosis in the progressive destruction of the lacrimal gland in the context of Sjögren's syndrome remains to be elucidated.
[0004] To date, immune cell infiltration is considered a major aspect of lacrimal gland involvement in Sjögren's syndrome. The pathogenic role of T cells, particularly helper T cell subsets, in Sjögren's syndrome is well-established, and their inflammatory cytokines are known to mediate tissue damage. The Janus kinase / signal transducer and activator of transcription pathway (JAK-STAT pathway) is an important downstream signaling cascade of key cytokines and participates in driving autoimmune pathology.
[0005] While current treatments for dry eye are diverse and have achieved some success, several limitations remain. Artificial tears require frequent, long-term use, placing high demands on patient compliance and safe storage. Anti-inflammatory drugs have relatively limited indications, and classic corticosteroids are unsuitable for long-term, high-dose use due to potential side effects. Therefore, in-depth research into the pathological mechanisms of the lacrimal glands in Sjögren's syndrome-related dry eye and the development of targeted therapies for specific pathways hold promise for a multi-dimensional and synergistic approach to alleviating the progression of Sjögren's syndrome-related dry eye. Summary of the Invention
[0006] To address the technical problems existing in the prior art, this invention provides the application of an anti-interferon-γ neutralizing antibody in the preparation of a drug for the prevention and / or treatment of dry eye syndrome. The technical solution is as follows:
[0007] Application of anti-interferon-γ neutralizing antibodies in the preparation of drugs for the prevention and / or treatment of dry eye syndrome.
[0008] Optionally, the drug is administered via injection.
[0009] Optionally, the dry eye condition includes dry eye associated with Sjögren's syndrome.
[0010] Optionally, the amount of the anti-interferon-γ neutralizing antibody is 2-10 mg / kg.
[0011] And / or, the dosage of the interferon-γ neutralizing antibody is 5-6 mg / kg.
[0012] A pharmaceutical composition for the prevention and / or treatment of dry eye syndrome, the pharmaceutical composition comprising: an anti-interferon-γ neutralizing antibody or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients.
[0013] Optionally, the drug is administered via injection.
[0014] Optionally, the dry eye condition includes dry eye associated with Sjögren's syndrome.
[0015] Optionally, the amount of the anti-interferon-γ neutralizing antibody is 2-10 mg / kg.
[0016] And / or, the dosage of the interferon-γ neutralizing antibody is 5-6 mg / kg.
[0017] Optionally, the pharmaceutical composition further includes: a protective agent against dry eye damage.
[0018] Optionally, the anti-dry eye damage protective drug includes: an anti-inflammatory agent.
[0019] The beneficial effects of the technical solutions provided in the embodiments of the present invention include at least the following:
[0020] This invention marks the first discovery of the application of interferon-γ neutralizing antibodies in the treatment of dry eye, achieving significant technical results. By inhibiting the release of interferon-γ into the lacrimal gland, interferon-γ neutralizing antibodies effectively protect lacrimal gland solid cells and alleviate ferroptosis in lacrimal gland duct cells. In an MRL (myotrophic lateral sclerosis) model of Sjögren's syndrome, injection of interferon-γ neutralizing antibodies significantly improved dry eye symptoms, including reduced corneal defects and increased tear secretion. Interferon-γ neutralizing antibodies provide a novel targeting strategy for the treatment of Sjögren's syndrome-related dry eye and may become a potent and effective drug for the clinical treatment of dry eye in the future. Attached Figure Description
[0021] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0022] Figure 1 This is a diagram showing the results of corneal lissamine green staining and tear secretion provided in Example 1 of the present invention;
[0023] Figure 2 This is a TUNEL staining result image of the lacrimal gland provided in Embodiment 2 of the present invention; wherein, DAPI labels the cell nucleus, TUNEL labels apoptotic cells, CYP2F2 labels the luminal duct cells, Merge is a fluorescence merged image, and Enlarged is a local magnified image;
[0024] Figure 3 This is an immunofluorescence result image of the lacrimal gland provided in Example 3 of the present invention; wherein, DAPI labels the cell nucleus, CD4 labels possible CD4+ T cells, CYP2F2 labels the luminal duct cells, and Merge is a fluorescence merged image;
[0025] Figure 4 This is a flow cytometry result of cervical lymph nodes provided in Embodiment 4 of the present invention. Detailed Implementation
[0026] The technical solution of the present invention will now be described with reference to the accompanying drawings.
[0027] In embodiments of the present invention, words such as "exemplarily," "for example," etc., are used to indicate that something is an example, illustration, or description. Any embodiment or design described as "exemplary" in the present invention should not be construed as being more preferred or advantageous than other embodiments or designs. Specifically, the use of the word "exemplary" is intended to present the concept in a concrete manner. Furthermore, in embodiments of the present invention, the meaning expressed by "and / or" can be both, or either one.
[0028] In this invention, interferon-γ (IFN-γ) is an important subtype of the interferon family and a classic pro-inflammatory factor, mainly secreted by Th1 cells.
[0029] The purpose of this invention is to overcome the shortcomings of the prior art and provide an application of an anti-interferon-γ neutralizing antibody in the prevention and / or treatment of dry eye associated with Sjögren's syndrome.
[0030] To achieve the above objectives, the technical solution adopted by the present invention is as follows:
[0031] In a first aspect, the present invention provides the use of an anti-interferon-γ neutralizing antibody in the preparation of a drug for the prevention and / or treatment of dry eye syndrome associated with Sjögren's syndrome.
[0032] This invention focuses on lacrimal gland damage within the context of Sjögren's syndrome-associated dry eye, centering on immune cells and functional solid cells within the lacrimal gland. Previous studies often treated the solid cells of the lacrimal gland as a single entity, neglecting the fact that the lacrimal gland actually comprises a series of cell types throughout tear secretion, including tear-secreting acini, luminal ducts involved in secretion and transport, basal ducts responsible for ductal proliferation, and myoepithelial cells that compress and promote tear secretion. Previous articles have insufficiently elucidated the clearly damaged cell types in Sjögren's syndrome and have limited understanding of the interactions between immune cells and solid cells. In Sjögren's syndrome, we have discovered the crucial role of luminal duct cells, which exhibit significant ferroptosis and a close association with Th1 cells and their secreted IFN-γ. Interferon-γ, a classic pro-inflammatory cytokine primarily secreted by Th1 cells, is found to be a significant factor in damaging downstream lacrimal gland solid cells in Sjögren's syndrome-associated dry eye. Our research further elucidates the mechanism by which IFN-γ blockade alleviates lacrimal gland damage, providing new insights for understanding the pathways involved and developing multi-target, multi-drug synergistic strategies.
[0033] Through extensive experimental research, the inventors have discovered for the first time that interferon-γ neutralizing antibodies, administered via systemic injection, can effectively treat Sjögren's syndrome-related dry eye. These antibodies significantly inhibit corneal epithelial damage in a Sjögren's syndrome model, protect lacrimal duct cells, and reduce lacrimal JAK-STAT pathway activation and barbed wire formation. This invention experimentally verifies that interferon-γ neutralizing antibodies can effectively control the occurrence and development of Sjögren's syndrome-related dry eye, providing new theoretical support for the clinical application of interferon-γ neutralizing antibodies in its treatment.
[0034] As a preferred embodiment of the first aspect, the effective mouse dose of the anti-interferon-γ neutralizing antibody is 50 mg / kg, which translates to approximately 5.6 mg / kg for human use, and is preferably administered via systemic injection. Because the anti-interferon-γ neutralizing antibody has a highly effective inhibitory effect on interferon-γ, a lower effective dose can be used to achieve its therapeutic or preventative effect on dry eye syndrome.
[0035] In a second aspect, the present invention provides a pharmaceutical composition for the prevention and / or treatment of dry eye syndrome associated with Sjögren's syndrome, the pharmaceutical composition comprising: an anti-interferon-γ neutralizing antibody or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
[0036] Animal experiments showed that in the Sjögren's syndrome model mice of this invention, the experimental group injected with 50 mg / kg of interferon-γ neutralizing antibody, compared with the control group, effectively protected corneal epithelial cells from the abnormalities in the quality and quantity of tear secretion caused by lacrimal gland inflammation, reduced corneal defects, and restored tear secretion. These results demonstrate its effective ocular surface protection effect.
[0037] As a preferred embodiment of the second aspect, when administered to animals, the effective dose of the interferon-γ neutralizing antibody is 50 mg / kg, preferably administered by systemic injection. The human dose of the interferon-γ neutralizing antibody is 2-10 mg / kg, preferably 5-6 mg / kg, and more preferably 5.6 mg / kg.
[0038] To make the technical problems, technical solutions and advantages of the present invention clearer, a detailed description will be given below in conjunction with the accompanying drawings and specific embodiments.
[0039] Unless otherwise specified, the experimental methods described in the following embodiments are conventional experimental methods well known to those skilled in the art, and are performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Where specific conditions are not specified in the experimental methods, they are generally operated under conventional conditions.
[0040] Unless otherwise specified, all materials and reagents described in the following examples are commercially available.
[0041] Example 1 Animal Experiment—Corneal Riesling with Lissamine Green and Tear Secretion Detection
[0042] 1. Animal drug administration and procedures
[0043] 1.1 The experimental animals were 6-8 week old female balb / c and MRL / lpr mice (purchased from Wuhan Shulaibao Biotechnology Co., Ltd.). The experiment was divided into: negative control group (balb / c (purchased from Wuhan Shulaibao Biotechnology Co., Ltd.), model control group (MRL / lpr mice (purchased from Wuhan Shulaibao Biotechnology Co., Ltd.), SS group), and experimental group (SS + 50mg / kg anti-interferon γ neutralizing antibody treatment group, SS + anti-IFNγ group).
[0044] 1.2 Interferon-γ neutralizing antibody was administered via intraperitoneal injection: Interferon-γ neutralizing antibody (purchased from Selleck (catalog number A2152)) was dissolved in 1×PBS solution. The initial dose per animal was 500 μg in 100 μL, followed by 250 μg every two days for a total of 1 mg. The control group received an equal volume of isotype antibody solution (purchased from Selleck (catalog number A2119)) via intraperitoneal injection.
[0045] 1.3 Mice were stained with lissamine green on the cornea at a specified date after drug administration, and the corneal epithelial staining was observed under a microscope. Tear volume was measured using phenol red cotton thread in the SS group and the SS+anti-IFNγ group. For specific procedures on lissamine green staining of the cornea and measurement of tear volume, please refer to ZENG H, YANG X, LIAO K, et al. Circadian disruptionreduces MUC4 expression via the clock molecule BMAL1 during dry eye development [J]. Experimental & Molecular Medicine, 2024, 56(7): 1655-66.
[0046] Experimental results:
[0047] Experimental results are as follows Figure 1 As shown.
[0048] from Figure 1 As shown in the middle left figure, the model control group showed a significant decrease in tear secretion compared to the negative control group; tear secretion was somewhat alleviated in the experimental group.
[0049] from Figure 1 As shown in the middle right figure, the ocular surface of the negative control group showed almost no corneal punctation; the model control group showed obvious positive staining of lissamine green, indicating significant corneal epithelial defects and obvious dry eye symptoms; the corneal epithelial defects in the experimental group were alleviated.
[0050] The above animal experiments show that, compared with the model control group, the small molecule compound eye drops in the experimental group of mice with Sjögren's syndrome can effectively prevent damage to corneal epithelial cells and restore tear secretion.
[0051] Example 2 Animal Experiment – TUNEL Staining of Lacrimal Gland
[0052] 1. Animal drug administration and procedures
[0053] 1.1 The experimental animals were 6-8 week old female balb / c and MRL / lpr mice. The experiment was divided into: negative control group (balb / c), model control group (SS, SS group), and experimental group (SS + 50mg / kg anti-IFNγ neutralizing antibody treatment group, SS + anti-IFNγ group).
[0054] 1.2 Interferon-γ neutralizing antibody was administered via intraperitoneal injection: The interferon-γ neutralizing antibody was dissolved in 1×PBS solution. The initial dose for each animal was 500 μg in 100 μL, followed by 250 μg every two days for a total of 1 mg. The control group received an equal volume of isotype antibody solution (purchased from Selleck (catalog number A2119)) via intraperitoneal injection.
[0055] 1.3 Mice in each group were euthanized by cervical dislocation on the designated date. The lacrimal glands were dissected, fixed, and embedded, followed by TUNEL staining. For specific procedures, please refer to CAO X, DI G, BAI Y, et al. Aquaporin5 DeficiencyAggravates ROS / NLRP3 Inflammasome-Mediated Pyroptosis in the Lacrimal Glands[J]. Invest Ophthalmol Vis Sci, 2023, 64(1): 4.
[0056] Experimental results:
[0057] Experimental results are as follows Figure 2 As shown.
[0058] from Figure 2 As can be seen, there was almost no FITC positive staining signal in the negative control group CYP2F2 positive cells; a large number of FITC positive staining signals appeared in the model control group CYP2F2 positive cells, indicating cell death; and the FITC positive staining signal was reduced in the experimental group CYP2F2 positive cells.
[0059] TUNEL staining in SS model mice indicated increased lacrimal duct cell death. However, in the experimental group of SS model mice treated with interferon-γ neutralizing antibodies, compared to the model control group, the interferon-γ neutralizing antibodies effectively alleviated lacrimal gland cell death.
[0060] Example 3 Animal Experiment—Immunofluorescence Staining of Lacrimal Gland
[0061] 1. Animal drug administration and procedures
[0062] 1.1 The experimental animals were 6-8 week old female MRL / lpr mice. The experiment was divided into two groups: model control group (SS, SS group) and experimental group (SS + 50mg / kg anti-interferon γ neutralizing antibody treatment group, SS + anti-IFNγ group).
[0063] 1.2 Interferon-γ neutralizing antibody was administered via intraperitoneal injection: The interferon-γ neutralizing antibody was dissolved in 1×PBS solution. The initial dose for each animal was 500 μg in 100 μL, followed by 250 μg every two days for a total of 1 mg. The control group received an equal volume of isotype antibody solution (purchased from Selleck (catalog number A2119)) via intraperitoneal injection.
[0064] 1.3 Mice in each group were euthanized by cervical dislocation on the designated date. The lacrimal glands were dissected, fixed, and embedded, followed by immunofluorescence staining. For specific procedures, please refer to YANG X, ZUO X, ZENG H, et al. IFN-gammaFacilitates Corneal Epithelial Cell Pyroptosis Through the JAK2 / STAT1 Pathway in Dry Eye [J]. Invest Ophthalmol Vis Sci, 2023, 64(3): 34.
[0065] Experimental results:
[0066] Experimental results are as follows Figure 3 As shown.
[0067] from Figure 3 As can be seen, in the model control group, there were obvious CD4 positive cells infiltrating around CYP2F2 positive ductal cells in the lacrimal gland, indicating that there may be obvious Th1 cell infiltration around the lacrimal gland solid cells and interaction between them. In the experimental group, after treatment with anti-interferon-γ neutralizing antibody, the infiltration of CD4 positive cells was significantly reduced, suggesting its alleviating effect on lacrimal gland solid cell damage.
[0068] The above animal experiments show that, compared with the model control group, the experimental group of SS model mice treated with anti-interferon-γ neutralizing antibody can effectively alleviate the infiltration of lacrimal gland immune cells, indicating that it also has a therapeutic effect on the local lacrimal gland.
[0069] Example 4: Flow Cytometry Results of Cervical Lymph Nodes
[0070] 1. Animal drug administration and procedures
[0071] 1.1 The experimental animals were 6-8 week old female MRL / lpr mice. The experiment was divided into two groups: model control group (SS, SS group) and experimental group (SS + 50mg / kg anti-interferon γ neutralizing antibody treatment group, SS + anti-IFNγ group).
[0072] 1.2 Interferon-γ neutralizing antibody was administered via intraperitoneal injection: The interferon-γ neutralizing antibody was dissolved in 1×PBS solution. The initial dose for each animal was 500 μg in 100 μL, followed by 250 μg every two days for a total of 1 mg. The control group received an equal volume of isotype antibody solution (purchased from Selleck (catalog number A2119)) via intraperitoneal injection.
[0073] 1.3 Mice in each group were euthanized by cervical dislocation on the designated date, and cervical lymph nodes were collected for flow cytometry analysis. For specific procedures, please refer to LIU X, SU Y, HUANG Z, et al. Sleep loss potentiates Th17-cellpathogenicity and promotes autoimmune uveitis [J]. Clin Transl Med, 2023, 13(5): e1250.
[0074] Experimental results:
[0075] Experimental results are as follows Figure 4 As shown.
[0076] from Figure 4 The left-middle figure shows that the proportion of CD4+IFN-γ+Th1 cells (Q2) was downregulated after treatment with interferon-γ neutralizing antibodies. Figure 4 As can be seen from the right-middle figure, the therapeutic effect is significant.
[0077] The above animal experiments show that, compared with the model control group, the experimental group treated with anti-interferon-γ neutralizing antibodies in SS model mice can effectively improve immune infiltration in the draining cervical lymph nodes. Furthermore, the systemic effect of the neutralizing antibodies can alleviate the local immune activation in the eye.
[0078] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.
Claims
1. Application of anti-interferon-γ neutralizing antibodies in the preparation of drugs for the prevention and / or treatment of dry eye syndrome.
2. The application according to claim 1, characterized in that, The drug is administered via injection.
3. The application according to claim 1 or 2, characterized in that, The dry eye condition mentioned includes dry eye associated with Sjögren's syndrome.
4. The application according to claim 3, characterized in that, The dosage of the anti-interferon-γ neutralizing antibody is 2-10 mg / kg; And / or, the dosage of the interferon-γ neutralizing antibody is 5-6 mg / kg.
5. A pharmaceutical composition for the prevention and / or treatment of dry eye syndrome, characterized in that, The pharmaceutical composition comprises: an anti-interferon-γ neutralizing antibody or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients.
6. The pharmaceutical composition according to claim 5, characterized in that, The drug is administered via injection.
7. The pharmaceutical composition according to claim 5, characterized in that, The dry eye condition mentioned includes dry eye associated with Sjögren's syndrome.
8. The pharmaceutical composition according to claim 7, characterized in that, The dosage of the anti-interferon-γ neutralizing antibody is 2-10 mg / kg; And / or, the dosage of the interferon-γ neutralizing antibody is 5-6 mg / kg.
9. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutical composition also includes: a protective agent against dry eye damage.
10. The pharmaceutical composition according to claim 9, characterized in that, The anti-dry eye damage protective drugs include: anti-inflammatory agents.