Application of fenofibrate in treatment of Fuchs corneal endothelial dystrophy

Abstract

The invention relates to the technical field of biological medicine, in particular to application of fenofibrate in treatment of Fuchs corneal endothelial dystrophy. By using the fenofibrate medicine, the thickness of the cornea posterior elastic layer in the FECD pathogenesis process can be effectively reduced, the reduction of the cornea endothelial cell density, the cell damage and the like are reduced, and finally the decompensation of the cornea endothelial cells is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to application of fenofibrate in treating Fuchs corneal endothelial dystrophy. Background Art

[0002] Fuchs' endothelial corneal dystrophy (FECD), also known as guttate cornea, is an age-related complex genetic disease characterized by the formation of extracellular matrix deposits and apoptosis of corneal endothelial cells (CECs).

[0003] FECD is clinically divided into early-onset and late-onset types. Among them, early-onset FECD is relatively rare and is caused by a point mutation in the pathogenic gene Col8a2. Late-onset FECD has a higher incidence rate and is more common in people aged 50-60 years. It is a degenerative disease caused by the combined effects of gene mutation and environmental factors. The incidence rate in women is 3-4 times that in men, and the condition in women is more serious. Late-onset FECD is clinically characterized by corneal guttae, thickening of the corneal posterior elastic layer, and progressive loss of corneal endothelial cells.

[0004] The formation of corneal globule warts on the Descemet's membrane (DM) is the earliest and most typical pathological feature of FECD. CECs are attached to the Descemet's membrane, which is composed of two layers. The layer close to the corneal stroma is formed during the fetal period and is called the Anterior Banded Layer or Fetal Layer (ABL). The layer close to the endothelium is the Posterior Nonbanded Layer (PNBL) formed by the secretion of CECs, which thickens year by year with age. The thickness of the Descemet's membrane in FECD patients is about 4 times that of normal people. The Posterior Nonbanded Layer becomes thinner or disappears and is replaced by the abnormally formed collagenous banded layer (CBL). Some patients also form a loose fibrous layer under the corneal endothelium, and the corneal globule warts are attached to the collagenous banded layer or fibrous layer.

[0005] Human corneal endothelial cells are non-regenerative cells. When the density of corneal endothelial cells is less than 500 / mm 2Symptoms such as corneal edema, painful subepithelial bullae, corneal opacity, etc. may occur, and even lead to blindness in severe cases. Late-onset FECD is the main disease that causes corneal endothelial dysfunction. The traditional method is corneal endothelial transplantation. In recent years, it has been found that the corneal endothelium in the area of ​​corneal drop-shaped warts in the center of the cornea can be torn off and removed, and then the local application of ripasudil can promote corneal endothelial migration, but it cannot inhibit corneal endothelial damage. This is also the only effective drug currently used in clinical treatment of FECD.

[0006] Thickening of the Descemet's membrane and the formation of corneal guttate are the primary links in the pathogenesis of FECD and are the key factors leading to subsequent corneal endothelial damage, apoptosis, and decompensation. Currently, no studies have reported effective means to inhibit abnormal proliferation of the Descemet's membrane, nor are there any drugs that treat FECD from the perspective of lipid metabolism. Summary of the invention

[0007] The present invention found that abnormal corneal endothelial energy metabolism in FECD patients is correlated with extracellular matrix remodeling. By regulating the lipid metabolism of the corneal endothelium, the thickening of the corneal Descemet's membrane can be effectively inhibited, thereby protecting the corneal endothelial cells. Based on this, the present invention uses the regulation of the lipid metabolism process of the corneal endothelium as a target for treating corneal endothelial fibrosis and thickening of the Descemet's membrane, thereby reducing the trend of thickening of the Descemet's membrane, and reversing or reducing corneal endothelial apoptosis and shedding, thereby treating FECD from the source.

[0008] Therefore, the present invention provides the use of fenofibrate in treating Fuchs corneal endothelial dystrophy, specifically, by using fenofibrate to act on the PPARa target to regulate the lipid metabolism of the corneal endothelium, thereby treating corneal endothelial fibrosis and thickening of the Descemet's membrane in Fuchs corneal endothelial dystrophy. Experiments show that oral fenofibrate can inhibit the thickening of the Descemet's membrane after corneal endothelial decompensation, can significantly reduce corneal endothelial damage, and inhibit corneal endothelial apoptosis, thereby treating Fuchs corneal endothelial dystrophy at the source.

[0009] In order to achieve the above-mentioned object of the invention, the technical solution provided by the present invention is as follows:

[0010] The present invention provides application of fenofibrate in treating Fuchs corneal endothelial dystrophy.

[0011] In the present invention, the Fuchs corneal endothelial dystrophy is a late-onset disease.

[0012] In the present invention, the application method is oral administration of fenofibrate.

[0013] In the present invention, the application method is to prepare eye drops containing fenofibrate.

[0014] In the present invention, the fenofibrate reduces the thickening of the corneal Descemet's membrane and / or reduces the apoptosis and shedding of the corneal endothelium by regulating the lipid metabolism of the corneal endothelial cells.

[0015] In the present invention, the target of fenofibrate in regulating lipid metabolism of corneal endothelial cells is PPARa.

[0016] The present invention also provides the use of PPARa as a target in screening or developing treatment for Fuchs corneal endothelial dystrophy.

[0017] The present invention further provides a drug for treating Fuchs corneal endothelial dystrophy, characterized in that the drug contains a therapeutically effective amount of fenofibrate.

[0018] According to the present invention, fenofibrate can inhibit the thickening of the corneal Descemet's membrane and protect the corneal endothelial cells by regulating the lipid metabolism of the corneal endothelium.

[0019] Specifically, by using fenofibrate, the thickness of the corneal Descemet's membrane can be effectively reduced during the development of FECD, the reduction in corneal endothelial cell density and cell damage can be improved, and ultimately the decompensation of corneal endothelial cells can be improved. This method can treat Fuchs corneal endothelial dystrophy from the root without the need for surgery or immunosuppression. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1A-Figure 1C The corneal morphology and transparency of mice at different injury time points were observed by slit lamp photography;

[0021] Figure 2 It is a statistical graph of mouse corneal turbidity;

[0022] Figure 3A-3C This is an OCT schematic diagram of the corneal status of mice at different injury time points;

[0023] Figure 4 This is a statistical diagram of mouse corneal thickness detected by OCT;

[0024] Figure 5 This is a morphological picture of corneal endothelial cells at different injury time points in mice observed by intravital confocal microscopy;

[0025] Figure 6A-6C HE staining and PSR staining were used to observe the morphology of corneal stroma and corneal Descemet's membrane;

[0026] Figure 7 This is a transmission electron microscope experiment to observe the morphology of the mouse corneal Descemet's membrane;

[0027] Figure 8This is a statistical chart of the thickness of the Descemet's membrane of the mouse cornea detected by transmission electron microscopy. DETAILED DESCRIPTION

[0028] Hereinafter, the present invention will be described in detail according to exemplary embodiments, but the present invention is not limited to these embodiments. The present invention is embodied in the following various forms, but should not be construed as being limited to the exemplary embodiments set forth herein. Therefore, the detailed description and embodiments of the present invention will convey the scope of the present invention to those of ordinary skill in the art and are construed as being within the scope of the present invention.

[0029] The present invention first discovered that fenofibrate can regulate the lipid metabolism of corneal endothelium, reduce the thickness of corneal Descemet's membrane during the onset of FECD, and improve the reduction of corneal endothelial cell density and cell damage, and finally improve corneal endothelial cell decompensation. Specifically, the target of fenofibrate in regulating the lipid metabolism of corneal endothelial cells is PPARa.

[0030] The method of using the fenofibrate drug of the present invention is to add fenofibrate to the feed of mice at a ratio of 0.2% (2 mg / kg), and the mouse feed intake is calculated as 4.5 g per day. In some embodiments, fenofibrate can be used in the form of an eye drop preparation, and the single dosage of fenofibrate is 0.1 g, three times a day.

[0031] Example 1 Effect of oral administration of 0.2% fenofibrate on mouse corneal endothelial cells

[0032] 1-1 Experimental Methods

[0033] A diet containing 0.2% fenofibrate was prepared and the experimental mice were divided into a normal diet group and a 0.2% fenofibrate diet group. 750 J / cm 2 UVA irradiation, the left eye was covered during the irradiation process. The left eye was the control group, and the right eye was the UVA damage group.

[0034] After the injury experiment was completed, the mice were further divided into a normal diet-fed group and a diet-fed group containing 0.2% fenofibrate.

[0035] After inducing chronic corneal endothelial injury, the mice were observed for 1 week, 2 weeks, and 3 weeks, and slit lamp photography was used regularly to observe the transparency of the mouse cornea. One month after the injury, the mouse corneal thickness was detected by OCT, the mouse corneal endothelial morphology was observed by confocal microscopy, the corneal stroma thickness and corneal posterior Descemet's membrane morphology were observed by HE staining, and the mouse corneal posterior Descemet's membrane and other structures were observed by transmission electron microscopy.

[0036] 1-2 Experimental Results

[0037] Attached Figures 1A-1CThe damage to the mouse cornea in different experimental groups at one, two and three weeks after UVA damage is shown respectively. The experimental results show that the degree of corneal damage in mice was relatively stable three weeks after UVA damage, and the corneal opacity of the UVA irradiation group fed with ordinary feed was more serious than that of the UVA irradiation group fed with 0.2% fenofibrate.

[0038] Attached Figure 2 The corneal opacity of different experimental groups one month after UVA damage is also shown. The experimental results show that the corneal opacity of the UVA irradiation group fed with ordinary feed is more serious than that of the UVA irradiation group fed with 0.2% fenofibrate.

[0039] Attached Figures 3A-3C The anterior segment OCT diagrams of the mouse cornea in different experimental groups at one, two, and three weeks after UVA damage are shown respectively. The experimental results show that the degree of corneal edema in the group fed with 0.2% fenofibrate was significantly reduced compared with that in the group fed with ordinary feed.

[0040] Attached Figure 4 The corneal thickness of different experimental groups one month after UVA damage is shown. The experimental results show that the corneal thickness of the UVA irradiated group fed with ordinary feed is greater than that of the UVA irradiated group fed with 0.2% fenofibrate.

[0041] Attached Figure 5 The figure shows the schematic diagram of the in vivo confocal microscopy of the mouse cornea in different experimental groups three weeks after UVA damage. The mice in the UVA irradiated group fed with ordinary feed showed enlarged corneal endothelial cells, irregular cell morphology, and reduced cell density in the same field of view, while the UVA irradiated group fed with 0.2% fenofibrate showed no significant decrease in cell density and uniform cell size.

[0042] The above experimental results show that oral administration of 0.2% fenofibrate has protective and therapeutic effects on corneal endothelial cells.

[0043] Example 2 Protection of fenofibrate against corneal damage

[0044] 2-1 Experimental Methods

[0045] A diet containing 0.2% fenofibrate was prepared and the experimental mice were divided into a normal diet group and a 0.2% fenofibrate diet group. 750 J / cm 2 UVA irradiation, the left eye was covered during the irradiation process. The left eye was the control group, and the right eye was the UVA damage group.

[0046] After the injury experiment was completed, the mice were further divided into a normal diet-fed group and a diet-fed group containing 0.2% fenofibrate.

[0047] One month after UVA damage, the mouse eyeballs were taken for paraffin sections for immunohistochemical staining, HE staining was used to observe cell damage, and Sirius (PSR) staining was used to observe changes in collagen in the cornea.

[0048] At 1 month after UVA damage, transmission electron microscopy experiments were performed to observe the changes in the Descemet's membrane.

[0049] 2-2 Experimental Results

[0050] Attached Fig. 6A The figure shows the changes in the corneal stroma morphology and corneal Descemet's membrane of mice in different experimental groups one month after UVA damage.

[0051] Among them, the experimental results of HE staining showed that after UVA damage, the corneal epithelium of mice became thinner (p < 0.001), the thickness of corneal endothelial cells became thinner and the density decreased, the cell nucleus staining deepened, and the matrix layer was significantly thickened, from about 60 microns to about 90 microns (p < 0.001), which far exceeded the corneal thickening caused by technical factors such as sectioning.

[0052] The results of HE staining also showed that the corneal epithelial cells of the mice fed 0.2% fenofibrate were thinner than those of the normal group, but the degree of corneal epithelial thinning in this group of mice did not differ before and after UVA irradiation. This may be related to factors such as weight loss caused by feeding fenofibrate, rather than UVA irradiation damage. Compared with the UVA irradiation group fed with ordinary feed, the corneal endothelial thickness of the UVA irradiation group fed with 0.2% fenofibrate was more preserved (p < 0.001), and there was no obvious shrinkage of the corneal endothelial cell nucleus, the thickness of the corneal endothelial cell layer was acceptable, and the thickness of the corneal stroma was greatly reduced (p < 0.001), indicating that the degree of corneal stroma edema was reduced. (See Figure 6B , 6C ).

[0053] In addition, the HE staining results also showed that the UVA-irradiated group fed with ordinary feed had a more obvious posterior elastic layer staining compared with the other groups, which is most likely related to the thickening of the posterior elastic layer. At the same time, after Sirius staining, we were able to observe that the UVA-irradiated group fed with ordinary feed had deeper and thicker staining of the posterior elastic layer after injury, but due to the limited microscope magnification, the thickness of the posterior elastic layer could not be statistically analyzed.

[0054] In order to further observe the changes in the elastic layer, a transmission electron microscope experiment was conducted. The experimental results are shown in the attached Figure 7 , Attachment Figure 8 As shown, the thickness of the Descemet's membrane of the cornea of ​​mice in the UVA-irradiated group fed with 0.2% fenofibrate did not change significantly compared with normal mice (about 2 μm), while the thickness of the Descemet's membrane of the UVA-irradiated group fed with ordinary feed increased by about 1 / 2 (about 3.0 μm).

[0055] The above experimental results show that oral fenofibrate can significantly inhibit the thickening of the corneal Descemet's membrane.

[0056] As determined in the above results, the present invention can effectively reduce the thickness of the corneal Descemet's membrane during the onset of FECD by using fenofibrate, improve the reduction of corneal endothelial cell density and cell damage, and ultimately improve corneal endothelial cell decompensation. This method can treat Fuchs corneal endothelial dystrophy from the root without surgery or immunosuppression.

[0057] It should be understood that the exemplary embodiments described herein should be considered in a descriptive sense only and not for limiting purposes. Descriptions of features or aspects in each embodiment should generally be considered to be feasible for similar features or aspects in other embodiments.

Claims

1. Application of fenofibrate in the preparation of drugs for the treatment of Fuchs' corneal endothelial dystrophy.

2. The use according to claim 1, characterized in that: The Fuchs corneal endothelial dystrophy is a late-onset disease.

3. The use according to claim 2, characterized in that: The application method is oral administration of fenofibrate.

4. The use according to claim 2, characterized in that: The application method is to prepare an eye drops preparation containing fenofibrate.

5. The use according to claim 2, characterized in that: The fenofibrate reduces the thickening of the corneal Descemet's membrane and / or reduces the apoptosis and shedding of the corneal endothelium by regulating the lipid metabolism of the corneal endothelial cells.

6. The use according to claim 5, characterized in that: The target of fenofibrate in regulating lipid metabolism of corneal endothelial cells is PPARa.

7. Application of PPARa as a target in screening or developing treatments for Fuchs' corneal endothelial dystrophy.

8. A drug for treating Fuchs corneal endothelial dystrophy, characterized in that The medicine contains a therapeutically effective amount of fenofibrate.

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