Application of meflunidone in preparation of medicine for treating posterior cataract

Abstract

The invention discloses an application of meflunidone in preparation of a medicine for treating posterior cataract. The meflunidone is applied to preparation of the medicine for treating the posterior cataract, a new treatment strategy is provided for the field of ophthalmology medical treatment, and the meflunidone has far-reaching influences on improvement of visual health and life quality of patients. Through scientific and rigorous experimental verification and clinical application, the invention shows the potential of the meflunidone in the aspect of preventing and treating PCO, and the meflunidone is applied to preparation of the medicine for treating posterior cataract to bring a new hope for cataract patients.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and in particular relates to an application of meflunisal in the preparation of a drug for treating post-cataract. Background Art

[0002] Cataract is the leading cause of blindness in the world. The global prevalence of cataract is 17.2%. It is the main cause of blindness in patients over 50 years old, and seriously endangers people's physical and mental health and quality of life. At present, there is no specific drug treatment for cataract, and surgery is still the most effective treatment. Posterior capsularopacification (PCO) is the main reason affecting the long-term effect of patients after surgery. It has been reported that the incidence of PCO in adults 10 years after cataract surgery can reach 40%, and the incidence of PCO in infants and young children 1 year after cataract surgery is almost 100%. The most commonly used treatment for PCO is Nd:YAG laser posterior capsulotomy, but laser treatment also has complications. Complications such as corneal endothelial damage, anterior chamber flare, anterior chamber hemorrhage, iritis, increased intraocular pressure, intraocular lens damage, macular edema, etc. can be seen after surgery. In severe cases, retinal holes and retinal detachment may even occur. After multiple laser treatments or even surgeries, PCO still recurs in children after cataract surgery, which undoubtedly increases the treatment risk and the economic burden of the children's families. Therefore, prevention of PCO remains a hot topic in the field of cataract research, especially for pediatric cataract surgery.

[0003] During cataract surgery, the lens fibers and the central anterior lens capsule with lens epithelial cells (LECs) attached are removed, leaving the lens bag to fix the intraocular lens (IOL) implanted in the eye. However, surgery cannot completely remove all LECs. The remaining LECs proliferate, migrate, and undergo epithelial-mesenchymal transition (EMT), that is, they transform into myofibroblasts to produce fibrotic PCO. As the disease progresses, LECs differentiate into abnormal lens fibers that form Soemmering rings around the bag or dense Elschnig pearls on the surface of the posterior capsule. The occurrence of different types of PCO seriously impairs the visual quality of cataract patients after surgery.

[0004] Under typical physiological conditions, the levels of various antioxidants in the lens are significantly higher than those in the aqueous humor. However, after cataract surgery, the levels of inflammation-related proteins, biomarkers of oxidative stress, and cytokines in the aqueous humor are significantly increased. This environment exposes the remaining LECs to continuous oxidative stress, prompting their tight junctions to dissolve and initiate the EMT process. EMT can be activated by the inflammatory response after cataract surgery, with damaged ocular tissue releasing chemokines to attract immune system cells to clear damaged tissue and promote tissue repair. In the wound healing response, multiple cytokines such as tumor necrosis factor alpha (TNF-α), interleukin 1beta (IL-1β), and prostaglandins play a key role in regulating tissue responses. The wound healing response also shapes cellular behavior, leading to increased expression of EMT markers such as α-smooth muscle actin (α-SMA), vimentin, fibronectin (FN), and collagen type I (Col I), while decreased expression of the epithelial marker E-cadherin (E-Cad), thereby promoting the occurrence of PCO.

[0005] In view of the pathogenesis of PCO, current research on the prevention of PCO mainly focuses on reducing residual LECs or regulating their cell behavior. Given the potential impact of PCO on patients' vision and the limitations of existing treatments, it is particularly urgent and necessary to develop a new, non-invasive, simple, efficient and universal treatment method.

[0006] Mefunidone (MFD) is a new anti-fibrotic drug developed by the applicant team. Previous studies have found that MFD has an inhibitory effect on inflammation, oxidative stress and cell apoptosis, thereby delaying renal and pulmonary fibrosis. However, it has not yet been studied and applied in the treatment of secondary cataracts. Summary of the invention

[0007] The technical problem to be solved by the present invention is to overcome the deficiencies and defects mentioned in the above background technology and provide a use of meflunisal in the preparation of a drug for treating post-cataract.

[0008] In order to solve the above technical problems, the technical solution proposed by the present invention is:

[0009] Disclosed is a use of 1-(4-((3-(4-methylpiperazin-1-yl)propyl)amino)benzyl)-5-(trifluoromethyl)pyridin-2(1H)-one (referred to as meflunisal) in the preparation of a drug for treating after-cataracts.

[0010] In the above application, preferably, the drug further comprises a pharmaceutically acceptable carrier and / or excipient.

[0011] Compared with the prior art, the present invention has the following beneficial effects:

[0012] (1) The present invention finds that meflunisal can reduce inflammation and oxidative stress response after cataract surgery, inhibit EMT process and play a role in treating PCO, and can be used to prepare drugs for treating secondary cataracts.

[0013] (2) The present invention uses meflunisal in the preparation of a drug for treating post-cataracts. Direct intracameral injection of a drug solution containing meflunisal (MFD) after cataract surgery can inhibit the proliferation, migration and EMT of lens epithelial cells, thereby effectively treating PCO. The drug can significantly reduce the incidence of PCO after cataract surgery, providing patients with an option to avoid or at least postpone laser treatment.

[0014] (3) Through the application of the present invention, adult cataract patients can expect to enjoy a significantly improved quality of life after surgery, avoiding the need for further vision loss and repeated treatment caused by PCO; for children with cataract, the application of the present invention provides an early intervention method to ensure the permeability of the visual pathway, which is of great significance for promoting the normal development of children's visual function.

[0015] In summary, the present invention uses meflunisal in the preparation of drugs for treating secondary cataracts, which not only provides a new treatment strategy for the field of ophthalmology, but also has a profound impact on improving patients' visual health and quality of life. Through scientific and rigorous experimental verification and clinical application, the present invention demonstrates the potential of meflunisal in the prevention and treatment of PCO, and uses it in the preparation of drugs for treating secondary cataracts, bringing new hope to cataract patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Schematic diagram of mouse extracapsular lens removal surgery in an embodiment of the present invention;

[0017] Figure 2 The expression changes of α-SMA on the lens capsule of mice after extracapsular lens extraction combined with intracameral injection of meflunizone in the embodiment of the present invention;

[0018] Figure 3 The expression changes of FN on the lens capsule of mice after extracapsular lens extraction combined with intracameral injection of meflunizone in the embodiment of the present invention;

[0019] Figure 4The changes in the expression of genes related to EMT and epithelial cells in the capsule of mice after extracapsular lens extraction combined with intracameral injection of meflunizone in the embodiment of the present invention;

[0020] Figure 5 This is a representative result of HE staining of the capsular bag after extracapsular lens extraction combined with intracameral injection of meflunizone in mice in the embodiment of the present invention;

[0021] Figure 6 The present invention shows the changes in the expression of inflammatory and oxidative stress markers in the capsular bag of mice after extracapsular lens extraction combined with intracameral injection of meflunizone. DETAILED DESCRIPTION

[0022] In order to facilitate the understanding of the present invention, the present invention will be described more comprehensively and meticulously below in conjunction with the accompanying drawings and preferred embodiments of the present invention, but the protection scope of the present invention is not limited to the following specific embodiments.

[0023] Unless otherwise defined, all professional terms used below have the same meanings as those generally understood by those skilled in the art. The professional terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the scope of protection of the present invention.

[0024] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0025] Embodiment 1:

[0026] A use of meflunisal in preparing a drug for treating after-cataract.

[0027] In the in vivo experiment, in order to simulate the intraocular microenvironment after cataract surgery, mice underwent extracapsular lens extraction (ECLE). Immediately after surgery, meflunisal solution was administered by intracameral injection. After evaluating the therapeutic effect, corneal and retinal toxicity, the appropriate intracameral injection concentration for mice was determined to be 50mmol / L. After intracameral injection of meflunisal, it was found that the proliferation and migration of lens epithelial cells in the anterior capsule were significantly reduced, the number of lens epithelial cells on the posterior capsule was significantly reduced, and no obvious thickening of the fibrous membrane was formed on the posterior capsule. At the same time, meflunisal inhibited the expression of fibrosis, inflammation, and oxidative stress markers, and inhibited the decreased expression of the epithelial marker E-cad, indicating that meflunisal has a therapeutic effect on PCO.

[0028] The specific test process includes:

[0029] 1. Experimental method: A mouse extracapsular lens extraction model was constructed to observe the efficacy of meflunisal in the treatment of PCO. Figure 1 As shown in the figure, 8-week-old male C57BL / 6 mice were fixed on the operating table and divided into the following two groups: ECLE combined with intracameral injection of PBS solution group (control group) and ECLE combined with intracameral injection of MFD solution group (experimental group). Three mice were repeated for each experiment, and the experiment was repeated three times independently. The eyeball was fixed with toothed forceps, and a 15-degree ophthalmic scalpel was used to make a corneal incision and cut the anterior capsule. The lens nucleus and cortex were separated and separated into the anterior chamber, delivered from the corneal incision, and the residual lens cortex was removed. The corneal incision was sutured with 10-0 sutures for 2 stitches. 31G insulin was used to inject 5μl of PBS solution (right eye) and 50mmol / L of meflunisal solution (left eye). After the operation, tobramycin dexamethasone eye ointment was applied to the eye, and it was properly placed in a cage box, paying attention to warmth and breathing, and waiting for its anesthesia recovery.

[0030] Lens capsular bag separation: 24h, 48h, and 5d after surgery, the eye reactions of mice were observed under a slit lamp microscope, and the occurrence of endophthalmitis was closely monitored. Lens capsular bags were removed 5 days after surgery, with 6-8 capsular bags as a group. A 15-degree ophthalmic scalpel was used to make a limbal incision, and the cornea was cut along the incision. The capsular bag tissue was clamped with sterile toothless forceps and placed in a 1.5mL sterile EP tube for subsequent protein immunoblotting (Western blot, WB) and real-time fluorescence quantitative PCR (Real-time fluorescence quantitative PCR, RT-qPCR) detection.

[0031] Frozen section and paraffin section sampling: Anesthetize the mouse. After anesthesia, cut the upper and lower eyelids of the mouse, cut the muscle tissue and optic nerve along the periphery of the eyeball, and be careful not to damage the posterior pole of the eyeball to ensure the integrity of the eyeball. After the eyeball is completely removed, rinse it in clean saline to remove residual blood and prepare frozen sections for immunofluorescence staining (IF) or paraffin sections for hematoxylin-eosin staining (HE).

[0032] 2. Experimental results

[0033] 2.1 Immunofluorescence staining results

[0034] After ECLE, the EMT markers α-SMA and FN increased significantly, but intracameral injection of meflunisal significantly inhibited the upregulation of EMT markers after ECLE (see Figure 2-Figure 3 ), which indicated that meflunisal could significantly inhibit the EMT process of LECs after ECLE (n=3).

[0035] 2.2WB and RT-qPCR results

[0036] RT-qPCR and WB were used to quantitatively verify the effect of intracameral injection of meflunisal on the expression of EMT-related markers in the lens capsular bag of mice after ECLE surgery, and the lens capsular bags of mice were collected 5 days after ECLE surgery for detection.

[0037] RT-qPCR and Western blot results are shown in Figure 4 As shown in the figure, 5 days after surgery, the EMT markers α-SMA, FN, Col I, and Vimentin in the lens capsule of the ECLE treatment group were significantly increased, and E-Cad was significantly decreased. However, meflunisal treatment inhibited the expression of EMT markers α-SMA, FN, Col I, and Vimentin, and restored the expression of epithelial marker E-Cad. This result is consistent with the results of immunofluorescence staining, and further confirmed at the mRNA and protein levels that meflunisal has an inhibitory effect on the EMT process of lens epithelial cells in mice after ECLE surgery (n=3).

[0038] The lens capsule of mice was collected 24 hours after ECLE surgery for detection. The results of RT-qPCR and Western blot are shown in Figure 6 As shown in (A, B, and D), inflammatory markers S100 calcium-binding protein A9 (S100A9), granulocyte colony-stimulating factor (G-CSF), chemokine ligand 1 (CXCL1), and cyclooxygenase-2 (COX-2) in the lens capsular bag of the ECLE-treated group were significantly increased 24 hours after surgery, while meflunisal treatment inhibited the expression of inflammatory markers (n=3).

[0039] 2.3HE Results

[0040] like Figure 5 As shown, HE staining of the eyeball reveals the appearance of the capsular bag after ECLE, in which the lens cortex and nucleus were removed to simulate the intraocular environment after cataract surgery. Immediately after ECLE, residual LECs were observed to be located in the anterior capsule and equator, while the posterior capsule remained transparent. By day 5 after surgery, LECs proliferated significantly and began to migrate to the equator and posterior capsule regions. The number of cells in the central region of the posterior capsule increased significantly, accompanied by some cell aggregation. The use of meflunisal reduced the severity of PCO after ECLE in mice (n=3).

[0041] 2.4 Malondialdehyde (MDA) measurement results

[0042] MDA is a compound generated during lipid peroxidation and is widely recognized as a key indicator for detecting lipid peroxidation levels in cells and tissues. The results of multi-time point analysis of MDA levels showed that the MDA content in the capsule increased significantly 24 hours after ECLE surgery and reached a peak level. Figure 6As shown in C, the experimental data also revealed that after intracameral injection of meflunisal, the expression level of MDA was significantly reduced (n=3).

[0043] The above test results show that meflunisal can effectively treat the occurrence of secondary cataract induced by extracapsular lens extraction in mice. This shows that the application of meflunisal in the preparation of drugs for treating secondary cataract can treat secondary cataract.

Claims

1. A use of meflunisal in the preparation of a drug for treating after-cataract.

2. The use according to claim 1, characterized in that The medicine further comprises a pharmaceutically acceptable carrier and / or excipient.

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