Use of rosiglitazone in the preparation of a medicament for preventing and / or treating dry eye

Abstract

This invention discloses the application of rosiglitazone in the preparation of drugs for the prevention and / or treatment of dry eye, relating to the field of biomedical technology. The application of rosiglitazone in the preparation of drugs for the prevention and / or treatment of dry eye is a first-time discovery of its application in the treatment of dry eye, particularly in the preparation of drugs for treating dry eye associated with circadian rhythm disorders. Rosiglitazone increases the number of conjunctival keratinocytes by inhibiting the ACSL4-related pathway. Simultaneously, the expression level of GPX4 in conjunctival tissue increases, and ferroptosis is inhibited. In a time-lapse model, oral administration of 0.243 mg / kg / day of rosiglitazone inhibited inflammatory activation caused by circadian rhythm disorders, significantly improving dry eye symptoms, including reduced corneal defects and mucin deficiency. Rosiglitazone provides a novel targeting strategy for the treatment of dry eye associated with circadian rhythm disorders 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 use of rosiglitazone in the preparation of drugs for the prevention and / or treatment of dry eye syndrome. Background Technology

[0002] Dry eye is a common ocular surface disease affecting visual quality and quality of life. In recent years, lifestyle-related dry eye has received increasing attention. Circadian rhythm disruption and sleep-wake disorders are common types of sleep disorders, frequently seen in those experiencing jet lag and those working night shifts. Clinical studies have found that night shift workers have increased tear film instability and a higher risk of dry eye compared to healthy individuals. However, the pathogenesis, clinical manifestations, and effective interventions of circadian rhythm-related dry eye remain unclear. The pathogenesis of circadian rhythm-related dry eye differs from that of ordinary dry eye, primarily influenced by circadian rhythm factors such as sleep, eating, and light exposure. It regulates ocular surface tissue damage through molecular biological clock regulation, leading to dry eye. Therefore, treatment should focus on the molecular biological clock and its downstream regulatory pathways.

[0003] While the roles of the cornea and lacrimal glands in the pathogenesis of dry eye associated with circadian rhythm disruption have received widespread attention, the role of the conjunctiva in the pathogenesis of dry eye remains incompletely understood. The conjunctival epithelium, composed of basal cells, mucin-secreting keratinocytes, and goblet cells, not only serves as a physical barrier of the ocular surface but also maintains tear film stability through mucin secretion. In dry eye, conjunctival epithelial dysfunction manifests as squamous metaplasia, loss of functional keratinocytes, and release of inflammatory cytokines, directly leading to mucin deficiency and a vicious cycle that continuously exacerbates ocular surface inflammation. Therefore, a deeper understanding of the cellular and molecular changes within the conjunctival epithelium is crucial for elucidating the pathogenesis of dry eye and identifying new therapeutic targets.

[0004] The current clinical treatment system for dry eye syndrome still relies on three main pillars: artificial tear replacement, anti-inflammatory intervention, and anti-infective therapy. Artificial tears, as a first-line commonly used medication, primarily function to replenish the aqueous layer of the ocular surface, preventing further corneal damage caused by dryness. Meanwhile, with a deeper understanding of the pathogenesis of dry eye, the inflammatory response has been recognized as a fundamental pathological element of the disease, leading to the widespread clinical use of anti-inflammatory drugs such as immunosuppressants like cyclosporine, corticosteroids, and nonsteroidal anti-inflammatory drugs (NSAIDs). Furthermore, for dry eye associated with Demodex mite infestation, antibiotics such as metronidazole are also included in the treatment strategy.

[0005] While existing treatments are diverse and effective, they still have many limitations. Artificial tears require frequent, long-term use, posing challenges to patient compliance and the safety of drug preservation; anti-inflammatory drugs have narrow indications, and corticosteroids are not suitable for long-term, high-dose use due to significant side effects. More importantly, the aforementioned intervention strategies mostly focus on tear volume replenishment or corneal inflammation relief, neglecting the initiating role of the conjunctiva in the early activation of dry eye inflammation. Therefore, there is an urgent need to develop novel therapeutic targets for dry eye associated with circadian rhythm disorders, thereby providing more targeted intervention options for this type of dry eye. Summary of the Invention

[0006] To address the technical problems existing in the prior art, this invention provides the application of rosiglitazone in the preparation of drugs for the prevention and / or treatment of dry eye syndrome. The technical solution is as follows: The use of rosiglitazone in the preparation of drugs for the prevention and / or treatment of dry eye syndrome.

[0007] Optionally, the dry eye syndrome is circadian rhythm disorder-related dry eye syndrome.

[0008] Alternatively, the rosiglitazone may be administered orally.

[0009] Optionally, the dosage of rosiglitazone in the drug is 0.243 mg / kg / day.

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

[0011] Optionally, the dry eye syndrome is circadian rhythm disorder-related dry eye syndrome.

[0012] Alternatively, the rosiglitazone may be administered orally.

[0013] Optionally, the dosage of rosiglitazone in the drug is 0.243 mg / kg / day.

[0014] Optionally, the pharmaceutical composition may be in one or more of the following forms: tablets, capsules, and dispersible tablets.

[0015] Optionally, the pharmaceutical composition further includes: an anti-inflammatory agent; And / or, the anti-inflammatory agent includes one or more of glucocorticoids and nonsteroidal anti-inflammatory drugs.

[0016] The beneficial effects of the technical solutions provided in the embodiments of the present invention include at least the following: This invention discloses for the first time the application of the ACSL4-specific inhibitor rosiglitazone in the preparation of a drug for treating dry eye associated with circadian rhythm disorders, preferably administered orally at a typical dose of 3 mg / kg / day. Experiments showed that, compared with the control group in a jet lag model, the treated mouse group exhibited significant relief of corneal epithelial defects and mucin deficiency. This discovery provides a novel drug target and treatment strategy for the treatment of circadian rhythm disorder-related dry eye, demonstrating potential for clinical application.

[0017] This invention marks the first discovery of the application of the ACSL4-specific inhibitor rosiglitazone in the treatment of dry eye, achieving significant technical effects. In particular, this invention discloses for the first time the application of rosiglitazone in the preparation of drugs for treating dry eye associated with circadian rhythm disorders, preferably administered orally at a typical dose of 0.243 mg / kg / day. Rosiglitazone, by inhibiting the ACSL4-related pathway, increases the number of conjunctival keratinocytes. Simultaneously, the expression level of GPX4 in conjunctival tissue increases, and ferroptosis is inhibited. In a time-lapse model, oral administration of 0.243 mg / kg / day of rosiglitazone inhibits inflammatory activation caused by circadian rhythm disorders, significantly improving dry eye symptoms, including reduced corneal defects and mucin deficiency. Rosiglitazone provides a novel targeting strategy for the treatment of dry eye associated with circadian rhythm disorders and may become a potent and effective drug for the clinical treatment of dry eye in the future. Attached Figure Description

[0018] 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.

[0019] Figure 1 This is a schematic diagram of the results of sodium fluorescein staining and risperidone green staining of the ocular surface provided in Example 1 of the present invention; Figure 2 This is a schematic diagram of the conjunctival keratinocyte staining results provided in Embodiment 2 of the present invention; Figure 3 This is a schematic diagram of the protein immunoblotting results of conjunctival glutathione peroxidase 4 provided in Example 3 of the present invention; Figure 4 This is a schematic diagram of the staining results of conjunctival inflammatory factor S100 calcium-binding protein A9 provided in Example 4 of the present invention. Detailed Implementation

[0020] The technical solution of the present invention will now be described with reference to the accompanying drawings.

[0021] 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.

[0022] In this invention, rosiglitazone: CAS No.: 122320-73-4, molecular formula: C 18 H 19 N3O3S, molecular weight: 357.43.

[0023] The purpose of this invention is to overcome the shortcomings of the prior art and provide rosiglitazone and its applications, which has the effect of treating dry eye associated with circadian rhythm disorders.

[0024] To achieve the above objectives, the technical solution adopted by the present invention is as follows: In a first aspect, the present invention provides a small molecule compound, rosiglitazone.

[0025] Rosiglitazone is a specific ACSL4 inhibitor that directly binds to ACSL4 and inhibits its enzymatic activity, thereby suppressing the activation of the downstream ferroptosis pathway. The ACSL4-ferroptosis pathway involves factors such as abnormal death of conjunctival basal cells and lack of conjunctival keratinocytes in the pathogenesis of dry eye associated with circadian rhythm disorders.

[0026] Through extensive experimental research, the inventors have discovered for the first time that rosiglitazone protects the ocular surface by blocking this pathway. Rosiglitazone, administered orally, is effective in treating dry eye associated with circadian rhythm disorders. Rosiglitazone significantly inhibits ferroptosis of conjunctival basal cells induced by circadian rhythm disorders in a jet lag model, ultimately alleviating corneal epithelial defects and mucin deficiency. This invention experimentally verifies that rosiglitazone can effectively control the occurrence and development of dry eye associated with circadian rhythm disorders, providing new theoretical support for the clinical application of rosiglitazone in its treatment.

[0027] In a preferred embodiment of the first aspect, the effective dosage of rosiglitazone in the small molecule compound composition is 3 mg / kg / day, preferably administered orally. Because rosiglitazone has a highly effective inhibitory effect on the ACSL4-related pathway, a lower effective dose can be used to exert its therapeutic or preventative effect on dry eye syndrome.

[0028] In a second aspect, the present invention provides a pharmaceutical composition for the prevention and / or treatment of circadian rhythm disorder-related dry eye, said pharmaceutical composition comprising the small molecule compound composition described in the first aspect, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0029] 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.

[0030] As a preferred embodiment of the second aspect, the pharmaceutical composition is formulated in one or more of the following forms: tablets, capsules, and dispersible tablets.

[0031] Thirdly, the present invention provides the use of the small molecule compound composition described in the first aspect or the pharmaceutical composition described in the second aspect in the preparation of a medicament for the prevention and / or treatment of dry eye syndrome.

[0032] Animal experiments showed that in the jet lag model mice used in this invention, rosiglitazone, administered orally at 3 mg / kg / day, increased the number of conjunctival keratinocytes compared to the control group. Simultaneously, GPX4 expression in the conjunctival tissue increased, and ferroptosis was inhibited. Further analysis revealed that rosiglitazone could inhibit inflammatory activation caused by circadian rhythm disruption, ultimately alleviating corneal epithelial defects and mucin deficiency. These results demonstrate the potent ocular surface protective effect of rosiglitazone in vivo.

[0033] As a preferred embodiment of the third aspect, when administering the drug to animals, the effective dose of rosiglitazone in the small molecule compound composition is 0.243 mg / kg / day (calculated from the mouse dose according to common methods in the art), preferably administered orally.

[0034] 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.

[0035] 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.

[0036] Unless otherwise specified, all materials and reagents described in the following examples are commercially available.

[0037] Example 1 Animal Experiment - Ocular Surface Fluorescein Sodium Staining and Rieslingamine Green Staining

[0038] (1) Method

[0039] 1. Animal drug administration and procedures

[0040] 1.1 The experimental animals were 6-8 week old female C57BL / 6j mice (purchased from Guangdong Yaokang Biotechnology Co., Ltd.). The experiment was divided into: a negative control group (Ctr group), a model control group (time lag modeling + drug solvent, JL + Vehicle group), and an experimental group (time lag modeling + 3 mg / kg / d rosiglitazone treatment group, JL + ROSI group). The specific steps for constructing the model control group (time lag modeling + drug solvent) are as follows: To establish a circadian rhythm baseline, all animals underwent circadian rhythm synchronization training before the experiment. Mice were placed in a constant light-dark cycle environment. Light was turned on at 8:00 AM (time factor 0, ZT0) daily to simulate the daytime environment, and turned off at 8:00 PM (time factor 12, ZT12) to simulate the dark environment. The chronic time lag model was established using a cross-time zone flight simulation paradigm, with an 8-hour photoperiod phase shift every 72 hours. The specific operational procedure is as follows: After circadian rhythm synchronization training, the light start time is advanced by 8 hours to 0:00, and the light-dark cycle is set as light period 00:00-12:00 and dark period 12:00-24:00. After maintaining this for 3 days, the light start time is advanced again by 8 hours to 16:00, forming a new light-dark cycle (16:00-04:00 light / 04:00-16:00 darkness), which lasts for 3 days. Then, the light start time is advanced again by 8 hours, and so on, for a total of 30 days of intervention.

[0041] 1.2 Rosiglitazone was administered via gavage: Rosiglitazone was dissolved in corn oil, and each animal was given a gavage dose of 3 mg / kg, 100 μL, once daily for 30 consecutive days. The model control group was given an equal amount of corn oil via gavage.

[0042] 1.3 Mice were stained with sodium fluorescein and lissamine green on the ocular surface at designated dates after drug administration. The specific steps were as follows: On days 7, 15, and 30 after modeling, mice were induced into general anesthesia by intraperitoneal injection of 1% pentobarbital (50 mg / kg). A sodium fluorescein test strip was moistened with sterile saline, and the lower conjunctival sac of the mouse was gently touched. The eyelid was gently moved to allow the sodium fluorescein solution to distribute naturally, followed by rinsing with saline to remove residual dye. An animal slit-lamp microscope imaging system was used, with the light source adjusted to cobalt blue light, to observe and photograph the corneal epithelial staining of each mouse. The corneal stained area was quantitatively calculated using ImageJ software, i.e., corneal defect area ratio = sodium fluorescein-stained positive area / total corneal area × 100%. After sodium fluorescein staining, 1% lissamine green staining solution prepared in PBS was dripped into the lower conjunctival sac of the mouse. The eyelid was gently moved to allow the lissamine green solution to distribute naturally, followed by rinsing with saline to remove residual dye. Using an animal slit-lamp microscope imaging system, the light source was adjusted to white light to observe and photograph the corneal lissamine green staining of each mouse. The corneal lissamine green staining was scored according to a previously reported scoring system, dividing the cornea into upper, middle, and lower sections. Each section was scored from 0 to 3 points based on the size of the stained area, with a total score range of 0 to 9 points.

[0043] (2) Experimental results

[0044] Experimental results are as follows Figure 1 As shown. From Figure 1 As can be seen, the model control group exhibited significant corneal epithelial defects and mucin deficiency, which worsened with increasing modeling time. Animal experiments showed that, compared to the model control group, the experimental group treated with small molecule compounds in the jet lag model mice effectively prevented corneal epithelial cell damage and mucin deficiency.

[0045] Example 2 Animal Experiment - Detection of Conjunctival Keratinocytes by Staining

[0046] (1) Method

[0047] 1. Animal drug administration and procedures

[0048] 1.1 The experimental animals were 6-8 week old female C57BL / 6j mice. The experiment was divided into: negative control group (Ctr group), model control group (time difference modeling + drug solvent, JL + Vehicle group), and experimental group (time difference modeling + 3 mg / kg / d rosiglitazone treatment group, JL + ROSI group).

[0049] 1.2 Rosiglitazone was administered via gavage: Rosiglitazone was dissolved in corn oil, and each animal was given a gavage dose of 3 mg / kg, 100 μL, once daily for 30 consecutive days. The model control group was given an equal amount of corn oil via gavage.

[0050] 1.3 Mice in each group were euthanized by cervical dislocation on the designated date. After dissection of the eyeballs and eyelids, they were fixed and embedded, followed by immunofluorescence staining detection. The specific steps are as follows: After dewaxing the paraffin sections of the eyeballs, antigen retrieval was performed sequentially, followed by blocking, incubation with the target primary antibody overnight, washing, and incubation with fluorescent secondary antibody. The samples were then observed and photographed under a fluorescence microscope.

[0051] (2) Experimental results

[0052] Experimental results are as follows Figure 2 As shown. From Figure 2 The results showed a significant reduction in MUC4-labeled conjunctival keratinocytes in the jet lag model mice. However, in the experimental group treated with rosiglitazone via gavage, compared to the model control group, rosiglitazone effectively restored the number of conjunctival keratinocytes.

[0053] Example 3 Animal Experiment - Schematic diagram of Western blot results for conjunctival GPX4

[0054] (1) Method

[0055] 1. Animal drug administration and procedures

[0056] 1.1 The experimental animals were 6-8 week old female C57BL / 6j mice. The experiment was divided into: negative control group (Ctr group), model control group (time difference modeling + drug solvent, JL + Vehicle group), and experimental group (time difference modeling + 3 mg / kg / d rosiglitazone treatment group, JL + ROSI group).

[0057] 1.2 Rosiglitazone was administered via gavage: Rosiglitazone was dissolved in corn oil, and each animal was given a gavage dose of 3 mg / kg, 100 μL, once daily for 30 consecutive days. The model control group was given an equal amount of corn oil via gavage.

[0058] 1.3 Mice in each group were euthanized by cervical dislocation at the designated date, and conjunctival tissue was dissected and subjected to Western blot analysis.

[0059] (2) Experimental results

[0060] Experimental results are as follows Figure 3 As shown. From Figure 3 The results showed that the expression of GPX4 in the conjunctiva of jet lag model mice was significantly reduced, indicating the occurrence of ferroptosis. In vivo experiments demonstrated that, compared with the control group, rosiglitazone effectively restored the expression level of GPX4 in the conjunctiva of the jet lag model mice administered via gavage, thus inhibiting ferroptosis.

[0061] Example 4 Animal Experiment - Staining of Conjunctival Inflammatory Factor S100A9

[0062] (1) Method

[0063] 1. Animal drug administration and procedures

[0064] 1.1 The experimental animals were 6-8 week old female C57BL / 6j mice. The experiment was divided into: negative control group (Ctr group), model control group (time difference modeling + drug solvent, JL + Vehicle group), and experimental group (time difference modeling + 3 mg / kg / d rosiglitazone treatment group, JL + ROSI group).

[0065] 1.2 Rosiglitazone was administered via gavage: Rosiglitazone was dissolved in corn oil, and each animal was given a gavage dose of 3 mg / kg, 100 μL, once daily for 30 consecutive days. The model control group was given an equal amount of corn oil via gavage.

[0066] 1.3 Mice in each group were euthanized by cervical dislocation on the designated date. After dissecting the eyeballs and eyelids, they were fixed and embedded. Immunofluorescence staining was then performed. The specific steps were as follows: after dewaxing the paraffin sections of the eyeballs, antigen retrieval was performed sequentially, followed by blocking, incubation with the target primary antibody overnight, washing, and incubation with the fluorescent secondary antibody. The samples were then observed and photographed under a fluorescence microscope.

[0067] (2) Experimental results

[0068] Experimental results are as follows Figure 4 As shown. From Figure 4 The results showed that the expression of the inflammatory cytokine S100A9 in the conjunctiva of jet lag model mice was significantly increased. In vivo experiments demonstrated that, compared with the control group, rosiglitazone administered by gavage to the jet lag model mice effectively reduced S100A9 levels and alleviated ocular surface inflammation.

[0069] 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. Use of rosiglitazone 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 syndrome mentioned is dry eye syndrome related to circadian rhythm disorder.

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

4. The application according to claim 1, characterized in that, The dosage of rosiglitazone in the drug is 0.243 mg / 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 rosiglitazone and a pharmaceutically acceptable carrier thereof.

6. The pharmaceutical composition according to claim 5, characterized in that, The dry eye syndrome mentioned is dry eye syndrome related to circadian rhythm disorder.

7. The pharmaceutical composition according to claim 5, characterized in that, Rosiglitazone is administered orally.

8. The pharmaceutical composition according to claim 5, characterized in that, The dosage of rosiglitazone in the drug is 0.243 mg / kg / day.

9. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutical composition is available in one or more of the following forms: tablets, capsules, and dispersible tablets.

10. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutical composition further includes: an anti-inflammatory agent; And / or, the anti-inflammatory agent includes one or more of glucocorticoids and nonsteroidal anti-inflammatory drugs.

Images