WO2020091430A1 - Composition for prevention or treatment of macular degeneration - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for prevention or treatment of macular degeneration, containing fursultiamine or salts thereof. Fursulthiamine reduces the expression of HIF-1α which is increased in retinal pigment epithelial cells, and inhibits the growth of choroidal vascular endothelial cells. A pharmaceutical composition of the present invention containing fursultiamine or salts thereof can also be used as an agent for treatment of various neovascular eye diseases.

Description

Composition for preventing or treating macular degeneration

The present invention was made by the task number HI16C1501 under the support of the Ministry of Health and Welfare, and the research institute specializing in the task is the Korea Health Industry Development Institute, the research project name is "Leading-type specialized research and development project", and the research project title is "Diabetes cardiovascular complications treatment And Efficacy Evaluation System Development ", the main institution is Kyungpook National University Hospital, and the research period is 2016.04.01 ~ 2021.03.31.

In addition, the present invention was made by task number 2017R1D1A1B03027966 under the support of the Ministry of Education, the research institute specializing in the project is the Korea Research Foundation, the research project name is "Personal Basic Research Support Project for Science and Engineering", and the research project title is "Semaphorin 3A and Angiopoietin- Evaluation of the efficacy of regulating choroidal retinal neovascularization and vascular permeability by microRNA control targeting like 4 ", the main institution is Kyungpook National University Industry-Academic Cooperation Foundation, and the research period is 2017.06.01 ~ 2020.05.31

In addition, the present invention was made by task number 2019R1A2C1084371 under the support of the Ministry of Education, the research institute specializing in the above task is the Korea Research Foundation, the research project name is "Scientific Researcher Support Project", and the research project title is "Mitochondrial energy metabolism of immune cells Research on new treatment mechanism of age-related macular degeneration through reprogramming and inflammation activation control ", Organizing organization is Kyungpook National University Industry-Academy Cooperation Group, Research period is 2019.09.01 ~ 2024.02.29.

This patent application claims priority to Korean Patent Application No. 10-2018-0133677 filed with the Korean Intellectual Property Office on November 2, 2018, and the disclosure of the patent application is incorporated herein by reference.

In addition, this patent application claims priority to Korean Patent Application No. 10-2019-0136771 filed with the Korean Intellectual Property Office on October 30, 2019, and the disclosures of the patent application are incorporated herein by reference.

The present invention relates to a composition for preventing or treating macular degeneration. More particularly, the present invention relates to a composition for the prevention or treatment of macular degeneration comprising fursultiamine or salts thereof.

It is known that macular degeneration is an ocular disease that causes vision impairment due to degeneration in the macular area, and it is known that the pathogenesis is related to age, family history, race, and smoking. At the beginning of the onset, the field of vision is blurred and the sight of the nearby vision is distorted, and later blindness occurs.

Age-related macular degeneration (AMD) causes severe irreversible vision loss and is known to be the leading cause of blindness in people over 50. In epidemiological studies, the prevalence rate was reported to be 1.2% of the 52-64-year-old population in the United States, and the rate was reported to be higher at 20-37% for people over 75 years of age, and the prevalence was expected to increase gradually as the average age increased. do.

There are two types of age-related macular degeneration. The first is non-angiogenic age-related macular degeneration, which is the most common and accounts for 85% of all age-related macular degeneration. This type of dryness is characterized by grease and atrophic changes in the retinal pigment epithelium. The second is neovascular age-related macular degeneration, characterized by choroidal neovascularization. Choroidal neovascularization is a newly formed blood vessel that tends to leak blood and body fluids. This leads to the formation of lesions in which fibrous tissue proliferates in the retinal tissue and the photoreceptors are lost, and continues to cause severe and irreversible vision loss.

In particular, according to a basic epidemiologic study of age-related macular degeneration published in 2010, the prevalence of age-related macular degeneration is 2.92% for early degeneration and 0.19% for late degeneration, but non-neovascular macular degeneration among late macular degeneration is 3.9%. , All the rest was observed as angiogenesis, indicating that the frequency of neovascular macular degeneration is much higher than in foreign cases.

Treatment of neovascular age-related macular degeneration includes photodynamic therapy and injection of drugs into the eye to block vascular endothelial growth factors. However, several large multi-center clinical studies have shown better results than intravitreal vascular endothelial growth factor (VEGF) injections than photodynamic therapy.

Accordingly, various anti-vascular endothelial growth factor drugs have been developed through the treatment of neovascular age-related macular degeneration, and among them, a representative drug is Ranibizumab (Lanibizumab, Lucentis®), which inhibits activity by binding to extracellular vascular endothelial growth factor. . Ranibizumab has been reported to be an effective and safe treatment in patients with neovascular age-related macular degeneration through various clinical studies and is widely used worldwide.

However, as the lesion frequently recurs, the number of injections increases, which places a considerable burden on the patient. In some patients, the lesion does not improve despite the injection.

On the other hand, since vascular endothelial growth factor (VEGF) acts as a powerful vasodilator, it serves to maintain the blood circulation and relaxation of the coronary arteries of the heart. Because age-related macular degeneration patients are older and at higher risk for cardiovascular disease, vascular endothelial growth factor injections also pose a risk of serious side effects.

Accordingly, the present invention intends to propose a new therapeutic composition that reduces the risk of side effects and brings an effective treatment effect against macular degeneration.

The present inventors have made extensive efforts to develop a therapeutic agent for macular degeneration with reduced risk of side effects. As a result, the present inventors completed the present invention by confirming that fursultiamine decreases the expression of increased HIF-1α in retinal pigment epithelial cells and inhibits choroidal vascular endothelial cell growth.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating macular degeneration.

Another object of the present invention is to provide a food composition for preventing or improving macular degeneration.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating neovascular ocular disease.

Another object of the present invention is to provide a food composition for preventing or improving neovascular eye disease.

Another object of the present invention is to provide a method for treating macular degeneration.

Another object of the present invention is to provide a method for treating neovascular eye disease.

One aspect of the present invention relates to a pharmaceutical composition for preventing or treating macular degeneration, including fursultiamine or salts thereof.

The present inventors have made extensive efforts to develop a therapeutic agent for macular degeneration with reduced risk of side effects. As a result, the present inventors confirmed that fursulthiamine reduces the expression of increased HIF-1α in retinal pigment epithelial cells and inhibits choroidal vascular endothelial cell growth.

Greener sulfonic thiamine (fursultiamine; thiamine tetrahydrofurfuryl disulfide, TTFD) is a vitamin of the vitamin B ₁ activity for the treatment of thiamine deficiency, and thiamine disulfide derivative. Compared to vitamin B ₁ , it is better absorbed into cells and produces a large amount of co-carboxylase, which improves neurological dysfunction, myocardial metabolic disorders, etc. that are physiologically related to deficiency of vitamin B ₁ or metabolic disorders. It is known.

The causes of macular degeneration are age increase, family history, race, and smoking, but are mainly caused by age increase. In macular degeneration, a yellow deposit called drusen (accumulation of extracellular proteins and lipids) gradually accumulates in the macula (part of the retina) between the retinal pigment epithelium and the choroid.

Age-related macular degeneration (AMD) as a function of age increases is based on the degree (size and number) of drusen, which is early, intermediate, and late. It is divided into stages.

According to one embodiment of the present invention, the macular degeneration is age-related macular degeneration (AMD).

According to another embodiment of the present invention, the macular degeneration is late age-related macular degeneration (late AMD).

In late AMD, retinal damage occurs, resulting in symptomatic vision loss in addition to drusen. Late AMD is divided into dry AMD and wet AMD depending on the type of damage. Dry AMD is characterized by geographic atrophy and is non-angiogenic AMD. On the other hand, wet AMD is neovascular AMD (A neovascular AMD) in which choroidal neovascularization occurs.

According to another embodiment of the invention, the macular degeneration is neovascular age-related macular degeneration (Neovascular age-related macular degeneration, Neovascular AMD) or non-angiogenic age-related macular degeneration (Non-neovascular age-related macular degeneration, Non -neovascular AMD).

According to another embodiment of the present invention, the macular degeneration is neovascular age-related macular degeneration.

The pharmaceutical composition of the present invention has an effective amount of fursulthiamine or a salt thereof from 100 mg / 60 kg / day to 240 mg / 60 kg / day.

The fursulthiamine used as an active ingredient in the present invention may be used in itself or in the form of a salt, preferably in the form of a pharmaceutically acceptable salt.

The salt is preferably an acid addition salt formed by free acid.

The free acid may be an organic acid and / or an inorganic acid.

The organic acids include citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, metasulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, glutanoic acid and aspartic acid. It may be, but is not limited thereto.

The inorganic acid may be hydrochloric acid, bromic acid, sulfuric acid and phosphoric acid, but is not limited thereto.

The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier, in addition to fursulthiamine or a salt thereof.

The pharmaceutically acceptable carrier is commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, Polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, but is not limited thereto.

The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered orally or parenterally.

For parenteral administration, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, ocular administration, or topical ocular administration may be used.

Topical administration of the eye, for example, is administered directly intraocularly, around the eyeball, around the eyeball, subretinal, central retinal, outside the fovea, subconjunctival, intravitreal ( intravitreous, intracameral or suprachoroidal.

The pharmaceutical composition of the present invention may also be administered through an insertion device.

Suitable dosages of the pharmaceutical compositions of the present invention vary by factors such as formulation method, mode of administration, patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, and sensitivity to radiation response. , Usually, a skilled physician can easily determine and prescribe a dose effective for the desired treatment.

The pharmaceutical composition of the present invention is prepared in a unit dose form by formulating using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily carried out by those skilled in the art to which the present invention pertains. Or it can be manufactured by incorporating into a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an ointment, ex-agent, powder, granule, tablet or capsule, and may further include a dispersant or stabilizer.

Another aspect of the present invention relates to a food composition for preventing or improving macular degeneration comprising fursulthiamine or a salt thereof.

When the composition of the present invention is a food composition, it may be prepared in the form of powder, granule, tablet, capsule or beverage.

In the present invention, the food may be candy, drink, gum, tea, vitamin complex, or health supplement food.

The food composition of the present invention may include fursultiamine or a salt thereof as an active ingredient, as well as ingredients commonly added in food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. It includes. Examples of the aforementioned carbohydrates include monosaccharides, such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, etc .; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.

As flavoring agents, natural flavoring agents (Tau Martin, Stevia extract (eg, rebaudioside A, glycyrrhizine, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

When the food composition of the present invention is prepared as a drink agent, in addition to fursultiamine or a salt thereof, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, peaworm extract, jujube extract, licorice extract, etc. may be additionally included. have.

Another aspect of the present invention relates to a pharmaceutical composition for preventing or treating neovascular ocular disease comprising fursulthiamine or a salt thereof.

Another aspect of the present invention relates to a food composition for preventing or improving neovascular eye disease comprising fursulthiamine or a salt thereof.

Since the pharmaceutical composition and food composition for preventing or treating neovascular ocular disease of the present invention include fursultiamine or a salt thereof, which is the same active ingredient as the pharmaceutical composition for preventing or treating macular degeneration of the present invention, between the two In order to avoid excessive complexity of the specification according to repeated description, the description is omitted.

The term "neovascular vascular disease" in the present specification is a pathological angiogenesis-related disease occurring in the eye, for example, corneal neovascularization, retinal neovascularization, choroidal neovascularization. , Including intraocular neovascularization, neovascular glaucoma, proliferative diabetic retinopathy, neovascular macular degeneration, and retinopathy of prematurity do.

Another aspect of the invention relates to a method for treating macular degeneration, comprising administering a pharmaceutical composition comprising fursulthiamine or a salt thereof to a subject in need thereof.

Another aspect of the present invention relates to a method of treating angiogenesis ocular disease, comprising administering a pharmaceutical composition comprising fursulthiamine or a salt thereof to a subject in need thereof.

The term "administration" as used herein means providing a substance to a subject in any suitable way. The route of administration of the pharmaceutical composition of the present invention can be administered orally or parenterally through all general routes as long as it can reach the target tissue. In addition, the pharmaceutical composition of the present invention may be administered using any device capable of delivering the active ingredient to target cells or organs.

The term “subject” herein is not particularly limited, for example, human, monkey, cow, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, mouse, rabbit, or Guinea pigs, preferably mammals, more preferably humans.

The features and advantages of the present invention are summarized as follows:

(a) The present invention relates to a pharmaceutical composition for preventing or treating macular degeneration, comprising fursultiamine or salts thereof.

(b) Pursulthiamine decreases the expression of increased HIF-1α in retinal pigment epithelial cells and inhibits choroidal vascular endothelial cell growth.

(c) The pharmaceutical composition comprising the fursulthiamine or a salt thereof of the present invention can also be used as a therapeutic agent for various neovascular eye diseases.

1 is a graph showing the effect of fursultiamine suppressing HIF-1α expression induced by hypoxic conditions in ARPE-19 cells.

Figure 2a shows the results of the choroid sprouting assay (choroid sprouting assay). The sprueing area was reduced by fursulthiamine. (Microscope magnification: 40 times)

2B is a graph showing quantitative analysis of the sprouting distance of FIG. 2A.

3 is a graph showing that in ARPE-19 cells, fursulthiamine inhibits increased VEGF secretion under hypoxic conditions.

4A-4B are results comparing vascular leakage through fluorescein angiography in a laser-induced choroidal neovascular model (laser-induced CNV). It can be seen that the degree of vascular leakage was decreased by fursulthiamine. 0: no laser spots, 1: no fluorescence brightness and size change between early and late, 2A: only fluorescence brightness between early and late changes, 2B: fluorescence brightness between early and late There is a change in both size and size.

5 shows that in a laser-induced choroidal neovascular model (laser-induced CNV), the size of CNV lesions by fursulthiamine is reduced. White bar: 100 μm.

Figure 6a is a graph of oxygen consumption change in the ARPE-19 cells, to confirm the energy metabolic changes of mitochondria.

6B is a spare capacity graph showing that in ARPE-19 cells, the metabolism of mitochondria, which was reduced by LPS, is recovered by fursulthiamine.

The present invention relates to a pharmaceutical composition for preventing or treating macular degeneration, including fursultiamine or salts thereof.

Hereinafter, the present invention will be described in more detail by the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Example 1: Effect of fursulthiamine on angiogenesis inhibition-HIF-1α reduction

Neovascular Age-related macular degeneration is characterized by choroidal neovascularization. It is known that the hypoxia environment of the retina or choroid expresses hypoxia inducible factor 1 alpha (HIF-1α), thereby promoting vascular endothelial cell factor to induce the development of new blood vessels in the choroid. The development of new blood vessels in the choroid eventually leads to visual impairment.

In the present invention, HIF-1α inhibition by the thiamine derivative fursultiamine and pursultiamine in the choroid to determine the effect of preventing the development of new blood vessels. In vitro experiments using retinal cell ARPE-19 (Adult Retinal Pigment Epithelial cell line-19) were performed.

ARPE-19 cells were dispensed in a 60 mm dish and allowed to adhere for about a day. The next day, 0 μM, 20 uM, 50 uM, and 100 uM of fursulthiamin hydrochloride (Toronto research chemical) and vehicle (Dimethyl sulfoxide (DMSO), Sigma-Aldrich) were treated by concentration in each cell. After 1 hour, they were placed in a hypoxic chamber (INVIVO ₂ 400, Baker) and exposed to 1% oxygen. And after 4-6 hours, the cells were taken out and treated with a protein lysis buffer to dissolve the cells, and then the proteins were separated.

After quantitative analysis of the protein using a BCA protein assay kit (Pierce BCA protein assay kit, Thermo Fisher Scientific), the sample was mixed with a sample of the same protein amount and 4X loading buffer and heated to denature the protein to a primary structure. The same amount of protein sample was loaded onto the SDS-PAGE gel and transferred to a PVDF membrane. The HIF-1a antibody (Novus, NB100-479) was diluted in a 5% BSA (bovine serum albumin) solution and incubated overnight at 4 ° C.

The next day, the membrane was incubated with a secondary antibody bound to HRP (Horseradish peroxidase), and reacted with an enhanced chemiluminescence (ECL) solution to induce chemiluminescence. Chemofluorescence image analysis equipment (GE Healthcare, LAS-4000) was used to detect the degree of luminescence according to the amount of protein to obtain an image. As a loading control gene, β-tubulin was used to show that the same amount of protein was loaded.

As a result, it was confirmed that the expression of HIF-1α increased by hypoxic conditions (1% oxygen) was decreased upon treatment with fursulthiamine (FIG. 1).

Example 2: Inhibitory effect of fursulthiamine on angiogenesis-decreased choroidal vascular endothelial cell growth

Through the mouse choroid sprouting assay, an ex vivo model of age-related macular degeneration, the effect of fursulthiamine on choroidal vascular endothelial cell growth was confirmed.

The eyeballs of C57 black / 6J (C57BL / 6J) from Jackson Laboratory, 3 or 4 weeks of age, were removed to separate the choroid / sclera and cut to a size of 1 mm × 1 mm. Meanwhile, 300 ul of liquid meth gel (Becton Dickinson, BD matrigel) was dissolved in ice into a 24-well plate, and cut choroids / sclera were planted one by one. Subsequently, the mixture was placed in a 37 ° C incubator for 10 minutes to harden the meth gel, and then 500 ul of EGM medium (Lonza, Endothelial Growth Medium) was added.

Thereafter, the growth of vascular endothelial cells was induced in a 37 ° C incubator. The medium was changed once every two days, at which time the fursulthiamine hydrochloride was treated at a concentration of 20 uM or 50 uM. The growth of vascular endothelial cells growing in the choroid was confirmed by observing 3 to 5 days after the planting of the choroid / sclera. The distance from the tissue to the sprawled range was measured and averaged at a total of four locations using the ImageJ program. Statistical processing was indicated to be significant when the p value was 0.05 or less (p <0.05) using the Prism program.

As a result, it was confirmed that vascular endothelial cell growth was decreased by fursulthiamine (FIGS. 2A-2B and Table 1).

Pursulthiamine (uM) 0 20 50 Sprouting range (pixels) 398.417 ± 40.827 229.926 ± 28.348 126.713 ± 22.810

Example 3: Inhibitory effect of fursulthiamine on VEGF secretion

ARPE-19 cells were dispensed in a 60 mm dish and allowed to adhere for about a day. The next day, after replacing with serum-free medium, 0 μM, 50 uM, and 100 uM (Toronto research chemical) and vehicle (DMSO, Sigma-Aldrich) of fursulthiamine hydrochloride were treated for each cell by concentration. And placed in a hypoxic chamber (INVIVO ₂ 400, Baker) and exposed to 1% oxygen.

And after 12 hours, the medium was recovered, and the amount of VEGF secreted in the medium was measured with a human VEGF enzyme immunoassay kit (Enzyme linked immunoassay (ELISA), R & D systems). The measurement method and concentration calculation were performed according to the manual provided in the kit. Statistical processing was indicated to be significant when the p value was 0.05 or less (p <0.05) using the Prism program.

As a result, it was confirmed that fursulthiamine inhibits VEGF secretion increased by hypoxic conditions (1% oxygen) (FIG. 3 and Table 2).

- vehicle One 2 3 Oxygen conditions 21% O ₂ 1% O ₂ 1% O ₂ 1% O ₂ Pursulthiamine (μM) 0 0 50 100 VEGF (pg / mL) 86.6 ± 3.2 184.2 ± 12.7 163.4 ± 7.4 47.4 ± 4.6

Example 4: Comparison of the degree of vascular leakage through fluorescein angiography

To view drug efficacy in a laser-induced choroidal retinopathy model, an animal model of macular degeneration, 7-8 week old C57 black / 6J (C57BL / 6J) mice were used. Fursultiamine (Fursultiamine) administration group and control (Control) 10 were used, respectively, 50 mg / kg of fursultiamine was orally administered for 8 days from the day before laser irradiation to the week after laser irradiation. As a control, sterile distilled water used as a solvent was administered in the same manner.

Mice were anesthetized with avertin (Sigma-Aldrich), and pupils were expanded by administering an antiseptic (Midrinpi, Taejun Pharmaceutical) to both eyes. Argon laser (Oculight GL, IRIDEX) was irradiated to both eyes to produce 4 laser spots per eye.

One week later, after anesthesia of the mouse, fluorescein (AK-FLUOR 10%, Akorn) was administered to the abdominal cavity, and angiography images of the base of the retina and fluorescein were taken using a MICRON IV Basic System (Phoenix Research Labs).

After fluorescein injection, the degree of fluorescence brightness and size change between the two periods was measured by taking the degree of vascular leakage in the early phase seen within 3 minutes and the late phase seen within 7 minutes. Confirmed.

When both the fluorescence brightness and the size of the spots increased, 2B was increased, only 2A when the brightness was increased, 1 if there was no difference, and scored as 0 if no spot was generated, and the results were compared. The more spots scored with 2B, the higher the angiogenesis leakage. Statistical processing was indicated to be significant when the p value was 0.05 or less (p <0.05) using the Prism program.

As a result, in the choroidal retinopathy model, the degree of angiogenesis leakage was decreased in the late phase when pursulthiamine was treated, and thus, it was confirmed that the degree of angiogenesis was reduced by pursulthiamine (FIGS. 4A-4B).

Example 5: Laser-induced choroidal neovascularization model (laser-induced CNV), confirming the size of CNV lesions by fursulthiamine

To induce a laser-induced choroidal retinopathy model, anesthesia of a 7-8-week-old C57 black / 6J (C57BL / 6J) mouse with avertin (Sigma-Aldrich), and an antiseptic in both eyes (midrinpi, Taejun Pharmaceutical) The pupil was expanded by administering. Argon laser (Oculight GL, IRIDEX) was irradiated with both eyes to produce 4 laser spots per eye. Fursultiamine (Fursultiamine) administration group and control (Control) 10 were used, respectively, 50 mg / kg of fursulthiamine was administered orally for 8 days from one day before laser irradiation to one week after laser irradiation. As a control, sterile distilled water used as a solvent was administered in the same manner.

One week after laser irradiation, the mice were anesthetized, and after fluorescein angiography, the eyes were removed for tissue staining and fixed in 4% paraformaldehyde (4% PFA, EMS) for 30 minutes. After removing the cornea and lens, the retina and choroid were separated. The separated choroid was put into a blocking buffer (blocking buffer, 0.2% bovine serum albumin, 5% normal goat serum, 0.5% Triton X-100) and reacted for 1 hour. One of the vascular markers, an isolectin antibody (Isolectin IB4-Alexa Fluor 488, Invitrogen), was diluted in a blocking buffer, and then put into a tissue to react. After that, the choroid was flattened, and a mount solution (Mountant, Thermo scientific) was placed thereon to cover and stabilize the cover glass.

The next day, using a confocal microscope (LSM800, Zeiss) was taken at a magnification of 100 times. The size of each laser spot was quantified using an Image J program. Statistical processing was indicated to be significant when the p value was 0.05 or less (p <0.05) using the Prism program.

As a result, it was confirmed that CNV lesion size was reduced by fursulthiamine (FIG. 5 and Table 3).

- Control Pursulthiamine administered group Lesion size (10 ^-3 mm ² ) 23.9 ± 2.4 15.7 ± 1.0

Example 6: Confirmation of mitochondrial metabolism change by fursulthiamine in retinal pigment epithelium

Since neovascular age-related macular degeneration is known to be a major pathological mechanism, it was confirmed whether mitochondrial metabolism is restored by pursulthiamine treatment and the effect of suppressing inflammation. Since metabolic changes in mitochondria may cause inflammation or exacerbation, it is predicted that when fursulthiamine restores the metabolism of mitochondria, it can be applied to the prevention or treatment of neovascular eye diseases accompanying inflammation.

Specifically, ARPE-19 cells were dispensed into 96-well XF plates, and the medium was changed once every 2 days. On the 5th day from the day of planting the cells, LPS (Lipopolysaccharides, Sigma-Aldrich) was treated with 10 ug / ml, and on the 6th day, fursulthiamine was treated with 50 uM. On the 7th day, it was replaced with XF medium (Seahorse XF DMEM medium, Agilent) and placed in a 37 ° C, Non-CO ₂ incubator for 30 minutes.

Oligomycin 2 uM, FCCP 0.5 uM, Rotenone 2 uM, Antimycin A 2 as a mitochondrial electron transport inhibitor and an inhibitor of cell respiration to confirm the mitochondrial spare capacity uM was used for each step.

The intracellular oxygen consumption rate (OCR) was measured with a Seahorse XF analyzer (Seahorse XFe96 analyzer, Agilent). The measurement method was conducted according to the manual provided by the Seahorse XF96 analyzer (Agilent).

As a result, it was confirmed that mitochondrial spare capacity reduced by LPS was recovered upon treatment with fursulthiamine. From this, it was confirmed that the change in mitochondrial energy metabolism by fursulthiamine may be involved in the inflammation inhibitory effect (FIGS.

Control LPS treatment group) LPS + fursulthiamine treatment group OCR (%) 153.614 ± 36.221 96.3486 ± 46.440 189.438 ± 21.392

These results show that the inhibitory effect of fursulthiamine in the pathogenesis of neovascular age-related macular degeneration suggests that the treatment of a composition comprising fursulthiamine will have the effect of preventing and treating macular degeneration.

The present invention relates to a composition for preventing or treating macular degeneration. More particularly, the present invention relates to a composition for the prevention or treatment of macular degeneration comprising fursultiamine or salts thereof.

Claims (9)

A pharmaceutical composition for preventing or treating macular degeneration, including fursultiamine or salts thereof. The pharmaceutical composition of claim 1, wherein the macular degeneration is age-related macular degeneration (AMD). The pharmaceutical composition of claim 2, wherein the macular degeneration is late age-related macular degeneration (late AMD). According to claim 2, The macular degeneration is neovascular age-related macular degeneration (Neovascular age-related macular degeneration, Neovascular AMD), the pharmaceutical composition. The method of claim 1, wherein the pharmaceutical composition is intraocular, periocular, posterior, subretinal, central retinal, central fovea, subconjunctival, intravitreous. , Intracameral or suprachoroidal. The pharmaceutical composition. Food composition for preventing or improving macular degeneration, including fursultiamine or salts thereof. A pharmaceutical composition for the prevention or treatment of neovascular ocular disease, including fursultiamine or salts thereof. The method of claim 7, wherein the neovascular eye diseases are corneal neovascularization, retinal neovascularization, choroidal neovascularization, intraocular neovascularization, neovascular glaucoma. glaucoma), proliferative diabetic retinopathy, neovascular macular degeneration or retinopathy of prematurity. Food composition for preventing or improving neovascular ocular disease, including fursultiamine or salts thereof.

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