Application of LGK974 in the preparation of drugs for the prevention and / or treatment of dry eye syndrome

Abstract

This invention discloses the application of LGK974 in the preparation of drugs for the prevention and / or treatment of dry eye syndrome, relating to the field of biomedical technology. The application of LGK974 in the preparation of drugs for the prevention and / or treatment of dry eye syndrome. This invention is the first to discover the application of the Wnt-β-catenin pathway inhibitor LGK974 in the treatment of dry eye syndrome. LGK974, by inhibiting the Wnt-β-catenin pathway, can effectively protect meibomian gland acini and ducts, restoring normal lipid metabolism and secretion. In a NOD (Normally Ophthalmic Disorder) Sjögren's syndrome model, oral administration of 200-400 μg / kg / day of LGK974 significantly improved dry eye symptoms, including reduced corneal defects and increased tear secretion, alleviating morphological and functional damage to the meibomian glands. LGK974 provides a new targeted strategy for the treatment of dry eye syndrome, especially Sjögren's syndrome-related dry eye syndrome and meibomian gland dysfunction-related dry eye syndrome, and may become a potent and effective drug for the clinical treatment of dry eye in the future.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the application of LGK974 in the preparation of drugs for the prevention and / or treatment of dry eye syndrome. Background Technology

[0002] Dry eye is a common disease in the field of ocular surface disorders, with core symptoms including dryness, foreign body sensation, and decreased vision. It is characterized by high incidence and a significant clinical burden. Among these, dry eye related to Sjögren's syndrome has received close attention. Sjögren's syndrome is a systemic immune disease affecting exocrine glands such as the salivary and lacrimal glands. Dry eye caused by Sjögren's syndrome is characterized by severe symptoms, prominent signs, and poor treatment outcomes, making it a current challenge in clinical treatment. Previous views held that the core of dry eye was an abnormality in the quality or quantity of tear secretion, with inflammation as the underlying cause. Long-term chronic inflammation of the cornea leads to oxidative stress and cell death, damaging the integrity of the corneal epithelium. Therefore, the etiology of dry eye is complex and diverse, involving various components of the ocular surface. Damage to the lacrimal glands causes insufficient tear secretion or changes in tear protein composition; conjunctival damage causes impaired mucin secretion by goblet cells; and damage to the meibomian glands causes lipid layer defects and accelerated tear evaporation, ultimately leading to corneal damage. Abnormal tear secretion caused by lacrimal gland immune infiltration in relation to Sjögren's syndrome has received widespread attention. However, recent studies have also pointed out that factors such as the conjunctiva and meibomian glands may be involved in its pathogenesis. Further understanding will help to fully depict the pathogenesis and find targeted drug targets.

[0003] Current clinical treatment for dry eye primarily revolves around artificial tear replacement, anti-inflammatory therapy, and anti-infective therapy. Artificial tears are the most commonly used first-line medication in clinical practice, mainly used to replenish the ocular surface fluid and prevent further corneal damage caused by dryness. On the other hand, with a deeper understanding of the disease mechanisms, the inflammatory response is considered the underlying cause of dry eye, and anti-inflammatory drugs such as immunosuppressants like cyclosporine, corticosteroids, and nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used clinically to treat dry eye. Antibacterial drugs such as metronidazole are also used to treat dry eye associated with Demodex mite infestation.

[0004] While existing treatments offer a wide range of options and have achieved some success, limitations remain. Artificial tears require long-term, frequent use, posing challenges to patient adherence and the safe storage of the medication. Anti-inflammatory drugs have relatively narrow indications, and classic anti-inflammatory drugs like corticosteroids are unsuitable for large-volume, long-term use due to their side effects. Furthermore, these interventions primarily alleviate dry eye by controlling tear production or corneal inflammation, neglecting the interaction between meibomian gland dysfunction and dry eye. The meibomian glands are important lipid-secreting glands on the ocular surface; lipid coating helps prevent excessive tear evaporation. Reduced meibomian gland volume, acinar atrophy, or ductal keratinization are considered closely related to dry eye. Identifying the meibomian gland pathological mechanisms of Sjögren's syndrome-related dry eye and developing therapeutic drugs targeting specific pathways would be beneficial for multi-faceted relief of Sjögren's syndrome-related dry eye. Summary of the Invention

[0005] To address the technical problems existing in the prior art, this invention provides an application of LGK974 in the preparation of drugs for the prevention and / or treatment of dry eye syndrome. The technical solution is as follows:

[0006] The use of LGK974 in the preparation of drugs for the prevention and / or treatment of dry eye syndrome.

[0007] Optionally, the dry eye syndrome includes: Sjögren's syndrome-associated dry eye syndrome and / or meibomian gland dysfunction-related dry eye syndrome.

[0008] Optionally, the drug is administered orally.

[0009] Optionally, the dosage of LGK974 is 200-400 μg / kg / day.

[0010] A pharmaceutical composition for the prevention and / or treatment of dry eye syndrome, the pharmaceutical composition comprising an effective dose of LGK974 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0011] Optionally, the dry eye syndrome includes: Sjögren's syndrome-associated dry eye syndrome and / or meibomian gland dysfunction-related dry eye syndrome.

[0012] Optionally, the pharmaceutical composition further includes a protective agent against dry eye damage;

[0013] And / or, the anti-dry eye damage protective drug includes: an anti-inflammatory agent.

[0014] Optionally, the dosage of LGK974 is 200-400 μg / kg / day.

[0015] Optionally, the pharmaceutical composition may be in the form of one or more tablets or capsules;

[0016] And / or, the tablets include: dispersible tablets.

[0017] The beneficial effects of the technical solutions provided in the embodiments of the present invention include at least the following:

[0018] This invention discloses for the first time the application of LGK974 in the preparation of drugs for treating dry eye, particularly Sjögren's syndrome-related dry eye and meibomian gland dysfunction-related dry eye, preferably administered orally at a typical dose of 200-400 μg / kg / day. Experiments showed that, compared with the control group of Sjögren's syndrome models, the treated mice exhibited reduced corneal epithelial defects and improved tear secretion. Furthermore, LGK974 effectively alleviated meibomian gland dysfunction closely related to dry eye and significantly reduced meibomian gland lipid metabolism. This discovery provides a novel drug target and treatment strategy for the treatment of dry eye, especially Sjögren's syndrome-related dry eye and meibomian gland dysfunction-related dry eye, and has potential for clinical application.

[0019] This invention marks the first discovery of the application of the Wnt-β-catenin pathway inhibitor LGK974 in the treatment of dry eye syndrome, achieving significant technical results. LGK974, by inhibiting the Wnt-β-catenin pathway, effectively protects meibomian gland acini and ducts, restoring normal lipid metabolism and secretion. In a NOD (Normally Ophthalmic Disorder) Sjögren's syndrome model, oral administration of 200-400 μg / kg / day of LGK974 significantly improved dry eye symptoms, including reduced corneal defects and increased tear secretion, alleviating morphological and functional damage to the meibomian glands. LGK974 provides a novel targeted strategy for the treatment of dry eye syndrome, particularly Sjögren's syndrome-related dry eye and meibomian gland dysfunction-related dry eye, and may become a potent and effective drug for the clinical treatment of dry eye in the future. Attached Figure Description

[0020] 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.

[0021] Figure 1 This is a diagram of corneal lissamine green staining and tear secretion results provided in Example 1 of the present invention, wherein CT represents the control group, SS represents the model control group, and SS+LGK974 represents the experimental group;

[0022] Figure 2 This is an image showing the results of meibomian gland oil red staining provided in Example 2 of the present invention;

[0023] Figure 3This is a gross photograph of the meibomian glands provided in Embodiment 3 of the present invention, wherein CT represents the control group, SS represents the model control group, and LGK974 represents the experimental group;

[0024] Figure 4 This is an image of the meibomian gland immunofluorescence and LipidTOX staining results provided in Example 4 of the present invention. In this image, DAPI is used to label the cell nucleus, and LipidTOX is used to label the lipid content in the cells. The darker the staining, the higher the lipid content. KPT1 is used to label the pathological keratin ducts in the meibomian gland. The last column represents the combined fluorescence of the above images. Detailed Implementation

[0025] The technical solution of the present invention will now be described with reference to the accompanying drawings.

[0026] 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.

[0027] In this invention, LGK974 refers to a specific Wnt-β-catenin pathway inhibitor that can inhibit the release of upstream Wnt protein and reduce downstream cascade activation. Specific information about LGK974 is as follows:

[0028] CAS No.: 1243244-14-5;

[0029] Molecular formula: C 23 H 20 N6O;

[0030] Molecular weight: 396.44.

[0031] The purpose of this invention is to overcome the shortcomings of the prior art and provide a Wnt-β-catenin pathway inhibitor LGK974 and its application, which has the efficacy of treating dry eye associated with Sjögren's syndrome and meibomian gland dysfunction.

[0032] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0033] In a first aspect, the present invention provides the use of the small molecule compound LGK974 in the preparation of a medicament for the prevention and / or treatment of dry eye syndrome.

[0034] This invention is the first to discover that the Wnt-β-catenin pathway is involved in the pathogenesis of Sjögren's syndrome-related dry eye and meibomian gland dysfunction, including factors such as weakened acinar lipid metabolism and ductal keratinization. LGK974 exerts a protective effect on meibomian glands by blocking this pathway.

[0035] Through extensive experimental research, the inventors have discovered for the first time that LGK974, administered orally, can effectively treat dry eye syndrome, particularly Sjögren's syndrome-related dry eye and meibomian gland dysfunction-related dry eye. For example, LGK974 can significantly inhibit corneal epithelial damage in a Sjögren's syndrome model, protect the normal morphology and function of the meibomian glands, and alleviate conjunctival aging phenotypes. This invention experimentally verifies that LGK974 can effectively control the occurrence and development of Sjögren's syndrome-related dry eye and meibomian gland dysfunction, providing new theoretical support for the clinical application of LGK974 in its treatment.

[0036] As a preferred embodiment of the first aspect, animal experiments have shown that in Sjögren's syndrome model mice, compared with the control group, the experimental group orally administered 2.5 mg / kg / d LGK974 effectively protected corneal epithelial cells from the effects of weakened meibomian gland lipid metabolism and duct keratinization, reduced corneal defects, and restored tear secretion. These results demonstrate the powerful ocular surface protective effect of LGK974 in vivo.

[0037] As a preferred embodiment of the first aspect, the effective dose of LGK974 is 200-400 μg / kg / day (roughly equivalent to 277.5 μg / kg / day for a 60kg adult), preferably administered orally. Because LGK974 has a highly effective inhibitory effect on the Wnt-β-catenin pathway, a lower effective dose can be used to achieve its therapeutic or preventative effect on dry eye syndrome.

[0038] In a second aspect, the present invention provides a pharmaceutical composition for the prevention and / or treatment of dry eye syndrome. The pharmaceutical composition comprises LGK974 as described in the first aspect, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0039] As a preferred embodiment of the second aspect, the pharmaceutical composition is a pharmaceutical composition for the prevention and / or treatment of Sjögren's syndrome-associated dry eye and meibomian gland dysfunction dry eye.

[0040] As a preferred embodiment of the second aspect, the pharmaceutical composition further includes other protective agents against dry eye damage, such as anti-inflammatory agents.

[0041] As a preferred embodiment of the second aspect, the pharmaceutical composition is formulated in one or more of the following forms: tablets and capsules.

[0042] The tablets include dispersible tablets.

[0043] 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.

[0044] 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.

[0045] Unless otherwise specified, all materials and reagents described in the following examples are commercially available.

[0046] Example 1: Animal Experiment – ​​Corneal Riesling with Lissamine Green and Tear Secretion Detection

[0047] 1. Animal drug administration and procedures

[0048] 1.1 The experimental animals were 6-8 week old female balb / c and NOD / ShiLtj mice (purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.). The experiment was divided into: negative control group (balb / c (purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.)), model control group (NOD / ShiLtj, SS group), and experimental group (NOD / ShiLtj + 2.5mg / kg / d LGK974 treatment group, SS + LGK974 group).

[0049] 1.2 LGK974 was administered via gavage: LGK974 (purchased from Selleck) was dissolved in corn oil, and each animal was given a gavage dose of 5 mg / kg, 100 μL, every two days for 10 consecutive days. The control group was given an equal amount of corn oil via gavage.

[0050] 1.3 Mice treated with the drug underwent corneal irisamine green staining on a specified date, and the corneal epithelial staining was observed under a microscope. Tear volume in the DED group and the DED+LGK974 group was measured using phenol red cotton thread. For specific procedures, please refer to ZENG H, YANG X, LIAO K, et al. Circadian disruption reduces MUC4 expression via the clock molecule BMAL1 during dry eye development [J]. Experimental & Molecular Medicine, 2024, 56(7): 1655-66.

[0051] Experimental results:

[0052] Experimental results are as follows Figure 1 As shown.

[0053] from Figure 1 As shown in the middle left figure, the tear volume measurement results show that LGK974 can significantly increase tear secretion in the SS model.

[0054] from Figure 1 As shown in the middle right figure, CT represents the negative control group, which received no experimental treatment; the SS group consisted of mice with Sjögren's syndrome, exhibiting significant corneal epithelial defects. The corneal defects in the SS+LGK974 group were alleviated.

[0055] The animal experiment in Example 1 showed that, compared with the model control group, the small molecule compound effectively prevented damage to corneal epithelial cells and restored tear secretion in the experimental group of mice with Sjögren's syndrome who were given small molecule compound eye drops.

[0056] Example 2 Animal Experiment – ​​Detection of Meibomian Gland Oil Red Staining

[0057] 1. Animal drug administration and procedures

[0058] 1.1 The experimental animals were 6-8 week old female balb / c and NOD / ShiLtj mice (purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.). The experiment was divided into: negative control group (balb / c (purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.)), model control group (NOD / ShiLtj, SS group), and experimental group (NOD / ShiLtj + 2.5mg / kg / d LGK974 treatment group, SS + LGK974 group).

[0059] 1.2 LGK974 was administered via gavage: LGK974 was dissolved in corn oil, and each animal was given a gavage dose of 5 mg / kg, 100 μL, every two days for 10 consecutive days. The control group was given an equal amount of corn oil via gavage.

[0060] 1.3 Mice in each group were euthanized by cervical dislocation on the designated date. After dissection of the eyelids, they were fixed and embedded, and then subjected to Oil Red staining for detection. For specific procedures, please refer to TU M, LIU R, XUE J, et al. Urban Particulate MatterTriggers Meibomian Gland Dysfunction [J]. Invest Ophthalmol Vis Sci, 2024, 65(2): 8.

[0061] Experimental results:

[0062] Experimental results are as follows Figure 2 As shown.

[0063] from Figure 2 As shown in the middle left figure, lipid embolism formation occurred in the SS model, and the lipid blockage of the duct was alleviated after LGK974 was administered.

[0064] from Figure 2 As shown in the middle right figure, the SS model showed insufficient lipid secretion in the acinar cells, and the lipid metabolism and lipid storage in the acinar cells were improved after LGK974 administration.

[0065] In SS model mice, oil red staining of the meibomian glands revealed reduced acinar lipid secretion and ductal lipid blockage. However, in the experimental group of SS model mice administered LGK974 via gavage, compared to the model control group, LGK974 effectively restored normal meibomian gland acinar lipid secretion and ductal lipid excretion.

[0066] Example 3 Animal Experiment – ​​Gross Photography of the Meibomian Glands

[0067] 1. Animal drug administration and procedures

[0068] 1.1 The experimental animals were 6-8 week old female balb / c and NOD / ShiLtj mice (purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.). The experiment was divided into: negative control group (balb / c (purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.)), model control group (NOD / ShiLtj, SS group), and experimental group (NOD / ShiLtj + 2.5mg / kg / d LGK974 treatment group, SS + LGK974 group).

[0069] 1.2 LGK974 was administered via gavage: LGK974 was dissolved in corn oil, and each animal was given a gavage dose of 5 mg / kg, 100 μL, every two days for 10 consecutive days. The control group was given an equal amount of corn oil via gavage.

[0070] 1.3 Mice in each group were euthanized by cervical dislocation on the designated date. After dissecting the eyelids, photographs were taken under a gross microscope, and the gland area was counted. For specific procedures, please refer to YANG X, ZHONG X, LIN H, et al. Deletion of Fgfr2 in Ductal Basal Epithelium With Tamoxifen Induces Obstructive Meibomian Gland Dysfunction [J]. Invest Ophthalmol Vis Sci, 2024, 65(13): 36.

[0071] Experimental results:

[0072] Experimental results are as follows Figure 3As shown.

[0073] from Figure 3 As can be seen in the left and middle images, the SS model shows a significant loss and reduction in the proportion of meibomian glands, including those in the upper and lower eyelids, while the LGK974 model partially restores the proportion of meibomian glands.

[0074] from Figure 3 As shown in the middle right figure, the loss of meibomian glands in the upper and lower eyelids and the therapeutic effect of LGK974 were both significant in the SS model.

[0075] The SS model mice exhibited significant glandular loss in both the upper and lower eyelids.

[0076] Animal experiments in Example 3 showed that, compared with the model control group, LGK974 effectively improved the morphology of meibomian glands in the experimental group of SS model mice that were administered LGK974 by gavage.

[0077] Example 4: Immunofluorescence and LipidTOX staining of meibomian glands

[0078] 1. Animal drug administration and procedures

[0079] 1.1 The experimental animals were 6-8 week old female balb / c and NOD / ShiLtj mice (purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.). The experiment was divided into: negative control group (balb / c (purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.)), model control group (NOD / ShiLtj, SS group), and experimental group (NOD / ShiLtj + 2.5mg / kg / d LGK974 treatment group, SS + LGK974 group).

[0080] 1.2 LGK974 was administered via gavage: LGK974 was dissolved in corn oil, and each animal was given a gavage dose of 5 mg / kg, 100 μL, every two days for 10 consecutive days. The control group was given an equal amount of corn oil via gavage.

[0081] 1.3 Mice in each group were euthanized by cervical dislocation on the designated date. After dissection of the eyelids, they were fixed and embedded, and then subjected to immunofluorescence and LipidTOX staining. For specific procedures, please refer to GUO Y, ZHANG H, ZHAO Z, et al. Hyperglycemia Induces Meibomian Gland Dysfunction [J]. Invest Ophthalmol VisSci, 2022, 63(1): 30.

[0082] Experimental results:

[0083] Experimental results are as follows Figure 4 As shown.

[0084] from Figure 4 It can be seen that the proportion of KRT1-labeled keratinized ducts was upregulated in SS, and lipid blockage in the ducts affected the normal discharge of meibomian gland lipids. LGK974 alleviated the degree of duct keratinization and reduced the degree of lipid embolism formation.

[0085] In SS model mice, the meibomian glands exhibited significant increases in ductal keratinization, lipid plug formation, decreased acini ratio, and reduced lipid secretion in both the upper and lower eyelids.

[0086] Animal experiments in Example 4 showed that, compared with the model control group, LGK974 effectively alleviated ductal keratosis and lipid plug formation, improved lipid secretion of acini, and restored normal acini ratio in the experimental group of SS model mice that were administered LGK974 by gavage.

[0087] 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 LGK974 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 dry eye syndromes include: Sjögren's syndrome-associated dry eye syndrome and / or meibomian gland dysfunction dry eye syndrome.

3. The application according to claim 1, characterized in that, The drug is administered orally.

4. The application according to any one of claims 1-3, characterized in that, The dosage of LGK974 is 200-400 μg / kg / day.

5. A pharmaceutical composition for the prevention and / or treatment of dry eye syndrome, characterized in that, The pharmaceutical composition comprises an effective dose of LGK974 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 5, characterized in that, The dry eye syndromes include: Sjögren's syndrome-associated dry eye syndrome and / or meibomian gland dysfunction dry eye syndrome.

7. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutical composition also includes a protective agent against dry eye damage; And / or, the anti-dry eye damage protective drug includes: an anti-inflammatory agent.

8. The pharmaceutical composition according to claim 5, characterized in that, The dosage of LGK974 is 200-400 μg / kg / day.

9. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutical composition may be in one or more of the following forms: tablets and capsules; And / or, the tablets include: dispersible tablets.

Images