Pharmaceutical composition for preventing or treating eye diseases, comprising altiratinib or pharmaceutically acceptable salt thereof as active ingredient

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating eye diseases, the composition comprising altiratinib or a pharmaceutically acceptable salt thereof as an active ingredient. More specifically, the present invention has been found to inhibit the Wnt / β-catenin signaling pathway and have the effect of suppressing intraocular vascular leakage, and thus can be used as a therapeutic agent for eye diseases such as macular degeneration, retinal edema, and retinal degeneration.

Description

Pharmaceutical composition for preventing or treating ocular diseases comprising altilatinib or a pharmaceutically acceptable salt thereof as an active ingredient

[0001] The present invention relates to a pharmaceutical composition for preventing or treating ocular diseases, comprising altilatinib or a pharmaceutically acceptable salt thereof as an active ingredient.

[0002] Edema is an excessive accumulation of fluids, such as plasma, interstitial fluid, and transcellular fluid, between cells in the outer part of the blood vessels due to leakage of fluid from the blood vessels (i.e., due to vascular leakage).

[0003] Vascular leakage within the eye can occur for a variety of reasons. For example, in patients with hypertension, persistently elevated blood pressure can cause destruction of the blood-retinal barrier, leading to retinal edema caused by vascular leakage due to the damage to the blood-retinal barrier. The macula is sometimes damaged by macular edema (macula tumentia) following lens removal for cataract treatment.

[0004] Retinal edema refers to an abnormal accumulation of fluid within the retina itself, while retinal detachment (detachment) is characterized by abnormal fluid accumulation in the subretinal space, causing the retina to separate from the underlying retinal pigment epithelium. Retinal detachment (detachment) or central retinal edema (corneal plaque) can result in significant loss of vision, ultimately leading to irreversible blindness.

[0005] Existing treatments for glaucoma include invasive surgery, localized laser therapy, and intravitreal drug administration. Commercially available medications administered topically directly into the vitreous cavity typically require repeated administration every four to six weeks. This repeated intravitreal administration can cause discomfort, pain, and side effects.

[0006] To overcome these problems, research is needed into technologies that can reduce the inconvenience, pain, and side effects of medication.

[0007] The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating ocular diseases, which comprises Altiratinib or a pharmaceutically acceptable salt thereof as an active ingredient.

[0008] Another object of the present invention is to provide a composition for inhibiting the movement of ß-catenin to the nucleus, comprising Altiratinib or a pharmaceutically acceptable salt thereof as an active ingredient.

[0009] Another object of the present invention is to provide a health functional food composition for preventing or improving eye diseases, which comprises altilatinib or a food-wise acceptable salt thereof as an active ingredient.

[0010] To achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating ocular diseases, comprising Altiratinib or a pharmaceutically acceptable salt thereof as an active ingredient.

[0011] In addition, the present invention provides a composition for inhibiting the movement of ß-catenin to the nucleus, comprising Altiratinib or a pharmaceutically acceptable salt thereof as an active ingredient.

[0012] In addition, the present invention provides a health functional food composition for preventing or improving eye diseases, which comprises altilatinib or a food-wise acceptable salt thereof as an active ingredient.

[0013] The present invention relates to a pharmaceutical composition for preventing or treating eye diseases, comprising altiratinib or a pharmaceutically acceptable salt thereof as an active ingredient. More specifically, it has been confirmed that altiratinib inhibits Wnt / β-catenin signaling and has an effect of inhibiting intraocular vascular leakage, and thus can be utilized as a treatment for eye diseases such as macular degeneration, retinal edema, and retinal degeneration.

[0014] Figure 1 shows the results of screening for Wnt / β-catenin signaling inhibitors.

[0015] Figure 2 shows the chemical structure of altilatinib.

[0016] Figure 3 confirms the inhibitory effect of Wnt / β-catenin-dependent signaling by altiratinib.

[0017] Figure 4 confirms the activity of inhibiting the movement of β-catenin into the nucleus.

[0018] Figure 5 confirms the effect of altilatinib on inhibiting intraocular vascular leakage.

[0019] In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives can be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents can be propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Base materials for suppositories include witepsol, macrogol, Tween 61, cacao butter, laurin butter, and glycerogelatin.

[0020] According to one embodiment of the present invention, the pharmaceutical composition can be administered to a subject in a conventional manner via intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalational, topical, rectal, oral, intraocular or intradermal routes.

[0021] The dosage of the active ingredient according to the present invention may vary depending on the condition and weight of the subject, the type and degree of the disease, the drug form, the route and period of administration, and may be appropriately selected by a person skilled in the art, and the daily dosage may be 0.01 mg / kg to 200 mg / kg, preferably 0.1 mg / kg to 200 mg / kg, and more preferably 0.1 mg / kg to 100 mg / kg. Administration may be once a day or divided into several times, and the scope of the present invention is not limited thereby.

[0022]

[0023] In addition, the present invention provides a composition for inhibiting the movement of ß-catenin to the nucleus, comprising Altiratinib or a pharmaceutically acceptable salt thereof as an active ingredient.

[0024]

[0025] In addition, the present invention provides a health functional food composition for preventing or improving eye diseases, which comprises altilatinib or a food-wise acceptable salt thereof as an active ingredient.

[0026] The above health functional food may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.

[0027] In addition, it may contain fruit pulp for the production of natural fruit juice, synthetic fruit juice, and vegetable drinks. These ingredients may be used independently or in combination. Furthermore, the health functional food composition may be in the form of any one of meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, drink, alcohol, and vitamin complex.

[0028] In addition, the above health functional food may additionally contain food additives, and its suitability as a “food additive” is determined by the specifications and standards for the relevant item in accordance with the general provisions and general testing methods of the Food Additives Codex approved by the Ministry of Food and Drug Safety, unless otherwise provided.

[0029] Examples of items listed in the above "Food Additives Codex" include chemically synthesized products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, kohlrabi pigment, and guar gum; and mixed preparations such as sodium L-glutamate preparations, alkaline agents added to noodles, preservative preparations, and tar color preparations.

[0030] At this time, the content of the effective ingredient added to the food during the process of manufacturing the health functional food can be appropriately increased or decreased as needed, and preferably, it can be added so that it is included in an amount of 1 to 90 parts by weight per 100 parts by weight of the food.

[0031]

[0032] Hereinafter, to aid understanding of the present invention, examples and other embodiments will be described in detail. However, the following examples and other embodiments merely illustrate the content of the present invention and are not intended to limit the scope of the present invention. The examples and other embodiments of the present invention are provided to more fully explain the present invention to those of average skill in the art.

[0033] <Example 1> Screening for Wnt / β-catenin signaling inhibitors

[0034] To search for a physiologically active substance that inhibits the Wnt / β-catenin signaling pathway, a human embryonic kidney cell line, HEK293, stably expressing a TOPflash marker plasmid, in which the expression of firefly luciferase is regulated by β-catenin, and a plasmid expressing human Frizzled-1 (hFz-1) was established (HEK293 marker cells). Wnt3a-secreting L cells were purchased from ATCC (American Type Culture Collection, USA) and cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS), 120 μg / ml penicillin, and 200 μg / ml streptomycin. Wnt3a-CM (Wnt3a-conditioned medium) was prepared by culturing Wnt3a-secreting L cells in DMEM containing 10% [v / v] FBS (fetal bovine serum) for 4 days, collecting the cultured DMEM medium, and sterilizing it with a 0.22 μm filter. Afterwards, fresh DMEM (containing 10% [v / v] FBS) medium was added to the cells, and the cells were cultured for an additional 3 days. The medium, collected in the same manner, was combined with the Wnt3a-CM obtained above.

[0035] HEK293 labeled cell line was seeded at 2.6x10 in a 96-well plate. 5 The cells were seeded at 100 cells / ml and cultured for 24 hours. Wnt3a-CM and candidate substances were added to each well at different concentrations (5 and 10 μM, respectively) for 15 hours, and after 15 hours, firefly luciferase activity and cell viability were measured using a TECAN SPARK 10M microreader, and the change in luciferase activity versus cell viability was analyzed.

[0036] As a result, according to Figure 1, it was confirmed that the firefly luciferase activity increased by Wnt3a-CM treatment was significantly reduced by altiratinib. On the other hand, it was confirmed that other VEGFR2 inhibitors, sitravatinib and pamufetinib mesylate, did not significantly inhibit firefly luciferase activity at the same concentration. Subsequent experiments were conducted using altiratinib, which significantly reduced firefly luciferase activity.

[0037]

[0038] <Example 2> Confirmation of the Wnt / β-catenin signaling inhibition effect of altiratinib

[0039] To confirm the inhibitory effect of altiratinib on Wnt / β-catenin signaling, ARPE-19 cells (human retinal epithelial cells) were purchased from ATCC (American Type Culture Collection, USA) and cultured in DMEM (Dulbecco's modified Eagle's medium) containing 10% FBS, 120 μg / mL penicillin, and 200 μg / mL streptomycin. The degree of activation or inhibition of the Wnt / β-catenin signaling pathway by altiratinib was measured by TOPFlash activity.

[0040] As a result, according to Fig. 3, it was confirmed that TOPFlash activity activated by a medium containing Wnt3a was reduced by altiratinib.

[0041] In addition, cytoplasmic and nuclear proteins were extracted from ARPE-19 cells treated with 5 and 10 μM of Wnt3a-CM and altiratinib for 15 hours, respectively. The amount of intracellular β-catenin, which regulates CRT (β-catenin response transcription) of the Wnt / β-catenin pathway, was confirmed by Western blotting using a β-catenin antibody (BD Transduction Laboratories, USA) and the ECL system (Santa Cruze Biotechnology). To confirm the nuclear protein fraction, nuclear protein PARP-1 was used as a reference substance.

[0042] As a result, according to Fig. 4, the amount of β-catenin in the nucleus increased in cells treated with Wnt3a-CM, but the amount of β-catenin in the nucleus further decreased in the altiratinib-treated group, confirming the inhibition of the Wnt / β-catenin pathway in human retinal epithelial cells.

[0043]

[0044] <Example 3> Inhibitory effect of altiratinib on intraocular vascular leakage

[0045] Twenty-one male chinchilla rabbits weighing approximately 2.0-2.5 kg were obtained from Saeron Bio Co., Ltd., and their health was confirmed by observing general symptoms during a 7-day acclimatization period. Healthy animals were used for the test. After instilling a mydriatic agent (1% Midriacyl eye drop) into the right eye of the animals, the animals were anesthetized, and then irradiated with a laser (LIGHTLas 532, LIGHTMED, USA) at the conditions of 532 nm, power 150 mW, and duration 0.1 sec, creating six spots at approximately the 6 o'clock direction centered on the optic nerve to induce choroidal neovascularization (CNV). The experimental animals were divided into three groups, G1 to G3, and the test substances were administered at the concentrations and methods shown in Table 1 below 7 days before the induction of CNV by the laser. In Table 1 below, Eylea (Aflibercept) was used as a positive control. For intravitreal injections, 50 μL of the positive control (Eylea) was administered intravitreally into the right eye of the anesthetized animal using a syringe equipped with a 31-gauge needle. For ocular transplants, a separate device provided by the sponsor was used for administration.

[0046] Number of animals by group, administered substance, route of administration, dosage (mg / eye), amount of solution (μL / eye), number of administrations, G1M6 vehicle, intravitreal --- G2M6 Illira, intravitreal 250 single doses, G3M6 Altiratinib, intravitreal 0.21 ea single dose

[0047] G1: Induced control group, G2: Positive control group, G3: Ocular implantation* Ilia: The original solution at a concentration of 40 mg / mL was used.

[0048] Afterwards, fundus photography and fluorescein intensity analysis were performed to measure CNV. Fundus photography was performed on the day of CNV induction (Day 0) and on days 7, 14, and 21 after the start of administration. After instilling a mydriatic agent (1% Midriacyl eye drop) into the right eye of the animals, the animals were anesthetized, and approximately 1 mL of a 2% fluorescein sodium salt solution was injected into the intravenous vein. The images were taken using a fundus camera (TRC-50IX, TOPCON, Japan) within approximately 2 minutes. Fluorescence intensity analysis was performed to confirm retinal CNV and evaluate drug efficacy using retinal fluorescein fundus photographs. Image analysis was performed using ImageJ software (NIH, Bethesda, MD) to analyze the fluorescence intensity of the irradiated area. Assuming normality of the data, one-way ANOVA was used to test the significance between the test groups. If significance was confirmed, a post hoc test was performed using Dunnett's multiple comparison test. Statistical analysis was performed using Prism 7.04 (GraphPad Software Inc., San Diego, CA, USA), and a p value less than 0.05 was considered statistically significant.

[0049] As a result, according to Fig. 5, the retinal fluorescence intensity analysis results showed that the retinal fluorescence intensity level of the altiratinib-administered group was significantly lower than that of the vehicle control group on the 21st day after the start of test substance administration, and the retinal fluorescence intensity level of the altiratinib-administered group was not statistically different from that of the positive control group, the Eylea-administered group. In conclusion, it was found that intraocular implant administration of altiratinib could help inhibit choroidal neovascularization in the chinchilla rabbit CNV model induced by laser irradiation.

[0050]

[0051] The foregoing description of the present invention is for illustrative purposes only. Those skilled in the art will readily appreciate that modifications to other specific embodiments can be made without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

[0052] The scope of the present invention is indicated by the claims set forth below, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims

1. A pharmaceutical composition for preventing or treating ocular diseases, comprising Altiratinib or a pharmaceutically acceptable salt thereof as an active ingredient.

2. In claim 1, The pharmaceutical composition is characterized in that it inhibits Wnt / ß-catenin signaling.

3. In claim 1, The pharmaceutical composition is characterized in that it suppresses blood leakage within the eye.

4. In claim 1, The pharmaceutical composition is characterized in that it inhibits the movement of ß-catenin into the nucleus.

5. In claim 1, A pharmaceutical composition characterized in that the above eye disease is at least one selected from the group consisting of macular degeneration, macular edema, retinal degeneration, retinal edema, diabetic retinopathy (DR), central retinopathy, retinopathy of prematurity, choroidal neovascularization, macula tumentia, and glaucoma.

6. In claim 1, A pharmaceutical composition characterized in that the pharmaceutical composition comprises 0.01 to 20 parts by weight of altiratinib based on 100 parts by weight of the total pharmaceutical composition.

7. In claim 1, A pharmaceutical composition characterized in that the above pharmaceutical composition is a composition for topical administration to the eye.

8. In claim 7, A pharmaceutical composition characterized in that the composition for topical administration to the eye is formulated as an eye drop, ointment, ocular injection, or ocular implant.

9. A composition for inhibiting the movement of ß-catenin to the nucleus, comprising Altiratinib or a pharmaceutically acceptable salt thereof as an active ingredient.

10. A health functional food composition for preventing or improving eye disease, comprising altilatinib or a food-related acceptable salt thereof as an active ingredient.

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