Ophthalmic composition for cell protection from light-induced damage

Abstract

To provide an ophthalmic composition that can protect a cell from disorder due to a light beam.SOLUTION: An ophthalmic composition for protecting a cell from disorder due to a light beam contains at least one selected from the group consisting of a sulfite, a vitamin E and zinc sulfate.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an ophthalmic composition for protecting cells from damage by light rays.

Background Art

[0002] In recent years, with the spread of personal computers, smartphones, and LEDs, the exposure to blue light has been increasing. Since blue light has strong energy, it is known that exposure to blue light causes symptoms such as eye fatigue, eye pain, and eye inflammation (for example, Non-Patent Document 1).

Prior Art Documents

Non-Patent Documents

[0003]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] While it is predicted that the exposure to blue light will further increase in the future, the present inventors have found that if cells present in the eye can be protected from damage by blue light, it may lead to alleviation of each of the above-described symptoms. An object of the present invention is to provide an ophthalmic composition capable of protecting cells from damage by light rays such as blue light.

Means for Solving the Problems

[0005] The present inventors have found that at least one component selected from the group consisting of sulfites, vitamin Es, and zinc sulfate can protect corneal epithelial stem cells from damage by blue light.

[0006] The present invention provides, for example, the following inventions. [1] An ophthalmic composition for cell protection from light damage, containing at least one selected from the group consisting of sulfites and vitamin E. [1] The ophthalmic composition according to [1], wherein the light is blue light. [3] The ophthalmic composition according to [1] or [2], wherein the cell is a stem cell. [4] The ophthalmic composition according to [3], wherein the stem cell is a corneal epithelial stem cell. [5] A corneal tissue regenerating agent from light damage, containing at least one selected from the group consisting of sulfites and vitamin E. [6] A method for evaluating the protective effect of the stem cell from damage by the blue light, including the steps of irradiating the stem cell with the blue light and measuring the cell viability of the stem cell.

[0007] The present invention also provides, for example, the following inventions. [2-1] An ophthalmic composition for cell protection from light damage, containing at least one selected from the group consisting of sulfites, vitamin E, aminoethylsulfonic acid and its salts, flavin adenine dinucleotide and its salts, pyridoxine and its salts, neostigmine and its salts, naphazoline and its salts, aspartic acid and its salts, zinc sulfate, and vitamin A. [2-2] The ophthalmic composition according to [2-1], wherein the light is blue light. [2-3] The ophthalmic composition according to [2-1] or [2-2], wherein the cell is a stem cell. [2-4] The ophthalmic composition according to [2-3], wherein the stem cell is a corneal epithelial stem cell. [2-5] A corneal tissue regenerating agent for protecting against light damage, containing at least one selected from the group consisting of sulfites, vitamin Es, aminoethylsulfonic acid and its salts, flavin adenine dinucleotide and its salts, pyridoxine and its salts, neostigmine and its salts, naphazoline and its salts, aspartic acid and its salts, zinc sulfate, and vitamin As.

[0008] The present invention also provides, for example, the following inventions. [3-1] An ophthalmic composition for cell protection against light damage, containing at least one selected from the group consisting of sulfites, vitamin Es, and zinc sulfate. [3-2] The ophthalmic composition according to [3-1], wherein the light is blue light. [3-3] The ophthalmic composition according to [3-1] or [3-2], wherein the cell is a stem cell. [3-4] The ophthalmic composition according to [3-3], wherein the stem cell is a corneal epithelial stem cell. [3-5] A corneal tissue regenerating agent for protecting against light damage, containing at least one selected from the group consisting of sulfites, vitamin Es, and zinc sulfate.

Advantages of the Invention

[0009] According to the present invention, an ophthalmic composition capable of protecting cells from light damage can be provided.

Modes for Carrying Out the Invention

[0010] Hereinafter, modes for carrying out the present invention will be described in detail. However, the present invention is not limited to the following embodiments.

[0011] In this specification, unless otherwise specified, the unit of content “%” means “w / v%” and is synonymous with “g / 100 mL”.

[0012] [1. Ophthalmic Composition for Cell Protection against Light Damage] The ophthalmic composition according to this embodiment contains at least one selected from the group consisting of sulfites, vitamin Es, and zinc sulfate (also referred to as “component (A)”).

[0013] In this specification, “light rays” means light of all wavelengths including ultraviolet rays, visible light rays, infrared rays, etc. The ophthalmic composition according to this embodiment can be preferably used for protecting cells from damage caused by blue light (blue light) having a wavelength of 380 to 500 nm among light rays.

[0014] The cells that can be protected by the ophthalmic composition according to this embodiment from damage caused by light rays are not particularly limited as long as they are cells that may be damaged by irradiation with light rays. Examples include corneal epithelial cells and retinal epithelial cells. Further, the ophthalmic composition according to this embodiment is preferably used for protecting stem cells having the ability of self-renewal and the ability to differentiate into different types of cells. Examples of such stem cells include corneal epithelial stem cells.

[0015] [Component (A)] [Sulfites] Sulfites are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable.

[0016] Examples of sulfites include salts of sulfurous acid, salts of pyrosulfurous acid, etc. Examples of salts of sulfurous acid and salts of pyrosulfurous acid include salts with inorganic bases [for example, ammonium salts, alkali metals (sodium, potassium, etc.), alkaline earth metals (calcium, magnesium, etc.), salts with metals such as iron]. Specific examples of sulfites include sodium bisulfite, sodium pyrosulfite, sodium sulfite, potassium bisulfite, potassium pyrosulfite, potassium sulfite, calcium sulfite, ammonium sulfite, iron sulfite, etc.

[0017] As sulfites, sodium bisulfite, sodium pyrosulfite, and potassium pyrosulfite are preferable, and sodium pyrosulfite is more preferable.

[0018] Commercially available sulfites can also be used. The sulfites may be used alone or in combination of two or more.

[0019] In the ophthalmic composition according to this embodiment, the content of the sulfites is preferably 0.0001 to 0.5 w / v% in terms of the total amount of the ophthalmic composition, more preferably 0.0005 to 0.1 w / v%, and still more preferably 0.001 to 0.05 w / v%.

[0020] 〔Vitamins E〕 The vitamins E are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable. Specific examples of the vitamins E include, for example, tocopherol, tocotrienol and their derivatives, and their salts. Tocopherol and tocotrienol may be any of α-, β-, γ- and δ-, and may be either the d-form or the dl-form.

[0021] Examples of the derivatives of the vitamins E include esters with organic acids such as tocopherol acetate, tocopherol succinate, tocopherol nicotinate, tocopherol linolenate and the like.

[0022] Examples of the salts of the vitamins E include organic acid salts (lactate, acetate, butyrate, trifluoroacetate, fumarate, maleate, tartrate, citrate, succinate, malonate, methanesulfonate, toluenesulfonate, tosylate, palmitate, stearate, etc.), inorganic acid salts (for example, hydrochloride, sulfate, nitrate, hydrobromide, phosphate, etc.), salts with organic bases (for example, salts with organic amines such as methylamine, triethylamine, triethanolamine, morpholine, piperazine, pyrrolidine, amino acid, tripyridine, picoline, etc.), salts with inorganic bases (for example, ammonium salts, salts with alkali metals such as sodium and potassium, salts with alkaline earth metals such as calcium and magnesium, salts with metals such as aluminum, etc.).

[0023] Examples of vitamin E include d-α-tocopherol, dl-α-tocopherol, β-tocopherol, γ-tocopherol, δ-tocopherol, vitamin E acetate (e.g., tocopherol acetate), vitamin E nicotinate, vitamin E succinate, and vitamin E linolenate. Tocopherol acetate (e.g., d-α-tocopherol acetate, dl-α-tocopherol acetate, etc.) is more preferred.

[0024] The vitamin E may be either a natural product or a synthetic product. A commercially available product can also be used as the vitamin E. The vitamin E may be used alone or in combination of two or more.

[0025] In the ophthalmic composition according to this embodiment, the content of vitamin E is preferably 0.0001 to 0.5 w / v% based on the total amount of the ophthalmic composition, more preferably 0.0025 to 0.15 w / v%, and still more preferably 0.01 to 0.05 w / v%.

[0026] [[Zinc Sulfate]] Zinc sulfate is not particularly limited as long as it is pharmaceutically, pharmacologically (in pharmaceutical terms), or physiologically acceptable.

[0027] The content of zinc sulfate in the ophthalmic composition according to this embodiment is not particularly limited and is appropriately set according to the type and content of other formulation components, dosage form, etc. As the content of zinc sulfate, for example, based on the total amount of the ophthalmic composition, the total content of zinc sulfate is preferably 0.001 to 1 w / v%, more preferably 0.005 to 0.5 w / v%, and particularly preferably 0.01 to 0.25 w / v%.

[0028] In another embodiment, the ophthalmic composition according to the present embodiment contains at least one selected from the group consisting of sulfites, vitamin Es, aminoethylsulfonic acid and its salts, flavin adenine dinucleotide and its salts, pyridoxine and its salts, neostigmine and its salts, naphazoline and its salts, aspartic acid and its salts, zinc sulfate, and vitamin As (also referred to as “component (A’)”). The sulfites, vitamin Es, and zinc sulfate in the present embodiment are as described in [component (A)].

[0029] [component (A’)] [Aminoethylsulfonic acid and its salts] Aminoethylsulfonic acid (taurine) and its salts are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutically), or physiologically acceptable.

[0030] Examples of the salts of aminoethylsulfonic acid include salts with organic bases (for example, salts with organic amines such as methylamine, triethylamine, triethanolamine, morpholine, piperazine, pyrrolidine, tripyridine, picoline, etc.) and salts with inorganic bases [for example, ammonium salts, salts with metals such as alkali metals (sodium, potassium, etc.), alkaline earth metals (calcium, magnesium, etc.), and aluminum].

[0031] Aminoethylsulfonic acid is preferred as aminoethylsulfonic acid and its salts.

[0032] Commercially available products can also be used as aminoethylsulfonic acid and its salts. Aminoethylsulfonic acid and its salts may be used alone or in combination of two or more.

[0033] In the ophthalmic composition according to this embodiment, the content of aminoethylsulfonic acid and its salts is preferably 0.001 to 10 w / v%, more preferably 0.01 to 5 w / v%, and still more preferably 0.1 to 1.0 w / v%, based on the total amount of the ophthalmic composition, where the total content of aminoethylsulfonic acid and its salts is concerned.

[0034] [Flavin adenine dinucleotide and its salts] Flavin adenine dinucleotide (vitamin B2) and its salts are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable.

[0035] Examples of salts of flavin adenine dinucleotide include salts with inorganic bases and salts with organic bases, preferably alkali metal salts, and particularly preferably sodium salts (sodium flavin adenine dinucleotide).

[0036] In the ophthalmic composition according to this embodiment, the content of flavin adenine dinucleotide and its salts is preferably 0.001 to 1.0 w / v%, more preferably 0.01 to 0.5 w / v%, still more preferably 0.03 to 0.1 w / v%, and even more preferably 0.03 to 0.05 w / v%, based on the total amount of the ophthalmic composition, where the total content of flavin adenine dinucleotide and its salts is concerned.

[0037] [Pyridoxine and its salts] Pyridoxine (vitamin B6) and its salts are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable.

[0038] Examples of salts of pyridoxine include organic acid salts (such as lactate, acetate, butyrate, trifluoroacetate, fumarate, maleate, tartrate, citrate, succinate, malonate, methanesulfonate, toluenesulfonate, tosylate, palmitate, stearate, etc.) and inorganic acid salts (such as hydrochloride, sulfate, nitrate, hydrobromide, phosphate, etc.).

[0039] As pyridoxine and its salts, pyridoxine and its inorganic acid salts are preferred, and pyridoxine hydrochloride is more preferred.

[0040] Commercially available pyridoxine and its salts can also be used. Pyridoxine and its salts may be used alone or in combination of two or more.

[0041] In the ophthalmic composition according to this embodiment, the content of pyridoxine and its salts is preferably 0.001 to 1.0 w / v% based on the total amount of the ophthalmic composition, more preferably 0.01 to 0.5 w / v%, still more preferably 0.02 to 0.2 w / v%, and even more preferably 0.05 to 0.1 w / v% in terms of the total content of pyridoxine and its salts.

[0042] 〔Neostigmine and its salts〕 Neostigmine and its salts are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable. Examples of salts of neostigmine include neostigmine methylsulfate. As neostigmine and its salts, neostigmine methylsulfate is preferred.

[0043] Commercially available neostigmine and its salts can also be used. Neostigmine and its salts may be used alone or in combination of two or more.

[0044] In the ophthalmic composition according to this embodiment, the content of neostigmine and its salts is preferably 0.00005 to 0.05 w / v% based on the total amount of the ophthalmic composition, more preferably 0.0001 to 0.01 w / v%, still more preferably 0.0005 to 0.005 w / v%, and even more preferably 0.001 to 0.005 w / v% in terms of the total content of neostigmine and its salts.

[0045] [Naphazoline and its salts] Naphazoline and its salts are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable. Examples of the salts of naphazoline include naphazoline hydrochloride and naphazoline nitrate. Naphazoline hydrochloride is preferred as naphazoline and its salts. Naphazoline and its salts may be used alone or in combination of two or more.

[0046] In the ophthalmic composition according to this embodiment, the content of naphazoline and its salts is preferably about 0.00001 to 0.1 w / v% of the total content of naphazoline or its salts based on the total amount of the ophthalmic composition, more preferably about 0.0001 to 0.01 w / v%, and even more preferably about 0.0003 to 0.003 w / v%.

[0047] [Aspartic acid and its salts] Aspartic acid is a compound known as an acidic amino acid also called 2-aminobutanedioic acid. Aspartic acid and its salts are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable. Aspartic acid may be any of the L-form, D-form, and DL-form, but preferably the L-form.

[0048] Examples of the salts of aspartic acid include salts with inorganic bases (e.g., ammonium salts; salts with metals such as alkali metals (sodium, potassium, etc.), alkaline earth metals (calcium, magnesium, etc.), aluminum, etc.), salts with organic bases (e.g., salts with organic amines such as methylamine, triethylamine, triethanolamine, morpholine, piperazine, pyrrolidine, tripyridine, picoline, etc.). As aspartic acid and its salts, salts of aspartic acid with inorganic bases are preferred, salts of aspartic acid with alkali metals and alkaline earth metals are more preferred, and potassium aspartate, magnesium aspartate, and magnesium potassium aspartate are even more preferred. Aspartic acid and its salts may be used alone or in combination of two or more.

[0049] The content of aspartic acid and its salts in the ophthalmic composition according to this embodiment is not particularly limited and is appropriately set according to the types and contents of other compounding components, dosage form, etc. As the content of aspartic acid or its salts, for example, based on the total amount of the ophthalmic composition, the total content of aspartic acid or its salts is preferably 0.01 to 20 w / v%, more preferably 0.01 to 5.0 w / v%, and even more preferably 0.2 to 2.0 w / v%.

[0050] 〔Vitamins A〕 The vitamins A are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable. Specific examples of vitamins A include, for example, retinol (vitamin A1), 3-dehydroretinol (vitamin A2) and their derivatives, and their salts.

[0051] Examples of the derivatives of vitamins A include esters with monocarboxylic acids such as retinol palmitate, retinol acetate, retinol butyrate, retinol propionate, retinol octylate, retinol laurate, retinol oleate and retinol linolenate.

[0052] Examples of salts of vitamin A compounds include organic acid salts [e.g., monocarboxylic acid salts (such as acetate, trifluoroacetate, butyrate, palmitate, stearate, etc.), polycarboxylic acid salts (such as fumarate, maleate, succinate, malonate, etc.), oxycarboxylic acid salts (such as lactate, tartrate, citrate, etc.), organic sulfonate salts (such as methanesulfonate, toluenesulfonate, tosylate, etc.), etc.], inorganic acid salts (e.g., hydrochloride, sulfate, nitrate, hydrobromide, phosphate, etc.), salts with organic bases (e.g., salts with organic amines such as methylamine, triethylamine, triethanolamine, morpholine, piperazine, pyrrolidine, tripyridine, picoline, etc.), and salts with inorganic bases [e.g., ammonium salts; salts with metals such as alkali metals (sodium, potassium, etc.), alkaline earth metals (calcium, magnesium, etc.), aluminum, etc.].

[0053] As vitamin A compounds, derivatives of retinol are preferred, esters of retinol and monovalent carboxylic acids are more preferred, retinol palmitate and retinol acetate are even more preferred, and retinol palmitate is even more preferred.

[0054] As vitamin A compounds, synthetic products may be used, or extracts obtained from natural products (e.g., vitamin A oil, etc.) may be used. Vitamin A oil is a fatty oil obtained from animal tissues containing retinol, or a concentrate thereof, or a product obtained by appropriately adding vegetable oil thereto. Commercially available products can also be used as vitamin A compounds. Vitamin A compounds may be used alone or in combination of two or more.

[0055] In the ophthalmic composition according to this embodiment, the content of vitamin A compounds is preferably such that the total content of vitamin A compounds is 5,000 to 200,000 IU / 100 mL, more preferably 10,000 to 100,000 IU / 100 mL, still more preferably 15,000 to 60,000 IU / 100 mL, even more preferably 20,000 to 55,000 IU / 100 mL, even still more preferably 40,000 to 55,000 IU / 100 mL, and particularly preferably 45,000 to 55,000 IU / 100 mL, based on the total amount of the ophthalmic composition.

[0056] "IU" means the international unit determined by the method described in the method for quantifying vitamin A in the 17th revised Japanese Pharmacopoeia. For example, in each article of pharmaceuticals in the 17th revised Japanese Pharmacopoeia, in the case of retinol acetate, it is described that it contains 2.5 million units or more of vitamin A per 1 g, and in the case of retinol palmitate, it contains 1.5 million units or more of vitamin A per 1 g.

[0057] [[(A) component or combination of (A') components]] The ophthalmic composition according to this embodiment may contain a combination of two or more of the above-described (A) components or (A') components. Thereby, the cell protection effect from damage by light is more significantly exhibited. Specific examples of the combination of the (A) component or (A') components include, but are not limited to, combinations of sulfites (particularly sodium pyrosulfite) and vitamin A compounds (particularly retinol palmitate), combinations of sulfites (particularly sodium pyrosulfite) and vitamin E compounds (particularly tocopherol acetate), combinations of zinc sulfate and aminoethylsulfonic acid (taurine), combinations of zinc sulfate and vitamin E compounds (particularly tocopherol acetate), and the like.

[0058] The pH of the ophthalmic composition according to the present embodiment is not particularly limited as long as it is within a pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable range. The pH of the ophthalmic composition according to the present embodiment may be, for example, 4.0 to 9.5, preferably 4.0 to 9.0, more preferably 4.5 to 9.0, still more preferably 4.5 to 8.5, even more preferably 5.0 to 8.5, and particularly preferably 5.0 to 8.0.

[0059] The ophthalmic composition according to the present embodiment can be adjusted to an osmotic pressure ratio within a range acceptable to the living body, if necessary. An appropriate osmotic pressure ratio can be appropriately set according to the use, formulation form, method of use, etc. of the ophthalmic composition. For example, it can be 0.4 to 5.0, preferably 0.6 to 3.0, more preferably 0.8 to 2.2, and still more preferably 0.8 to 2.0. The osmotic pressure ratio is the ratio of the osmotic pressure of the sample to 286 mOsm (the osmotic pressure of a 0.9 w / v% aqueous sodium chloride solution) based on the 17th revised Japanese Pharmacopoeia, and the osmotic pressure is measured with reference to the osmotic pressure measurement method (cryoscopic method) described in the Japanese Pharmacopoeia. Note that the standard solution for measuring the osmotic pressure ratio (0.9 w / v% aqueous sodium chloride solution) is prepared by drying sodium chloride (Japanese Pharmacopoeia standard reagent) at 500 to 650 °C for 40 to 50 minutes, then allowing it to cool in a desiccator (silica gel), accurately weighing 0.900 g of it, dissolving it in purified water and making it exactly 100 mL, or a commercially available standard solution for measuring the osmotic pressure ratio (0.9 w / v% aqueous sodium chloride solution) can be used.

[0060] The viscosity of the ophthalmic composition according to the present embodiment is not particularly limited as long as it is within a pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable range. The viscosity of the ophthalmic composition according to the present embodiment is preferably such that, for example, the viscosity at 20 °C measured with a rotational viscometer (RE550 type viscometer, manufactured by Toki Sangyo Co., Ltd., rotor; 1°34’×R24) is 1 to 10000 mPa·s, more preferably 1 to 8000 mPa·s, still more preferably 1 to 1000 mPa·s, even more preferably 1 to 100 mPa·s, and particularly preferably 1 to 20 mPa·s.

[0061] The ophthalmic composition according to this embodiment may contain, in an appropriate amount, components selected from various pharmacologically active components and physiologically active components in addition to the above components, as long as the effects of the present invention are not impaired. The components are not particularly limited, and for example, the active ingredients in ophthalmic drugs described in "Guidelines for Approval of Manufacturing and Sales of Pharmaceuticals Requiring Guidance / General Use, 2017 (Supervised by the Regulatory Science Society of Japan)" can be exemplified. Specific examples of the components used in ophthalmic drugs include, for example, the following components. Anti-allergic agents: For example, sodium cromoglycate, tranilast, pemirolast potassium, etc. Anti-histamine agents: For example, iproheptine, diphenhydramine hydrochloride, chlorpheniramine maleate, ketotifen fumarate, olopatadine hydrochloride, levocabastine hydrochloride, etc. Amino acids: For example, monoaminomonocarboxylic acids such as glycine, alanine, aminobutyric acid, aminovaleric acid, aminocaproic acid; monoaminodicarboxylic acids such as aspartic acid, glutamic acid or their salts; diamino monocarboxylic acids such as arginine, lysine or their salts, etc. Steroid agents: For example, fluticasone propionate, fluticasone furancarboxylate, mometasone furancarboxylate, beclomethasone propionate, fluocinolide, etc. Decongestants: For example, tetrahydrozoline hydrochloride, tetrahydrozoline nitrate, epinephrine, epinephrine hydrochloride, ephedrine hydrochloride, phenylephrine hydrochloride, dl-methylephedrine hydrochloride, etc. Ocular muscle regulating agents: For example, cholinesterase inhibitors having an active center similar to acetylcholine, specifically, tropicamide, helenine, atropine sulfate, etc. Anti-inflammatory agents: For example, glycyrrhetinic acid, dipotassium glycyrrhizinate, pranoprofen, methyl salicylate, glycol salicylate, allantoin, tranexamic acid, ε-aminocaproic acid, berberine chloride, berberine sulfate, sodium azulene sulfonate, lysozyme, licorice, etc. Astringents: For example, zinc white, zinc lactate, etc. Fat-soluble vitamins (excluding vitamin E): For example, retinol palmitate, retinol acetate, etc. Water-soluble vitamins: For example, cyanocobalamin, panthenol, calcium pantothenate, sodium pantothenate, etc. Local anesthetics: For example, lidocaine, procaine, etc. Others: For example, sulfamethoxazole, sodium sulfamethoxazole, etc.

[0062] In the ophthalmic composition according to this embodiment, within a range that does not impair the effects of the present invention, various additives may be appropriately selected and contained in an appropriate amount by combining one or more thereof according to the usual method according to the use and dosage form. As such additives, for example, various additives described in the "Pharmaceutical Additives Dictionary 2007" (edited by the Japan Pharmaceutical Additives Association) can be exemplified. The following additives are listed as representative components. Carriers: For example, aqueous solvents such as water and hydrous ethanol. Chelating agents: For example, ethylenediaminediacetic acid (EDDA), ethylenediaminetriacetic acid, ethylenediaminetetraacetic acid (EDTA), N-(2-hydroxyethyl)ethylenediaminetriacetic acid (HEDTA), diethylenetriaminepentaacetic acid (DTPA), etc. Bases: For example, octyldodecanol, titanium oxide, potassium bromide, plastic base, etc. pH adjusters: For example, hydrochloric acid, acetic acid, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, triethanolamine, diisopropanolamine, etc. Surfactants: For example, nonionic surfactants such as tyloxapol, polyoxyethylene sorbitan fatty acid esters, polyoxyl stearate, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, poloxamers, etc.; anionic surfactants such as polyoxyethylene alkyl ether phosphates, polyoxyethylene alkyl ether sulfates, alkylbenzene sulfonates, alkyl sulfates, N-acyl taurine salts, etc.; zwitterionic surfactants such as lauryldimethylaminoacetic acid betaine, etc. Cooling agent: for example, menthol, menthone, camphor, borneol, geraniol, cineole, citronellol, carvone, anethole, eugenol, limonene, linalool, linalyl acetate, thymol, cymene, terpineol, pinene, camphene, isoborneol, fenchene, nerol, myrcene, myrcenol, linalyl acetate, lavandulol, eucalyptus oil, bergamot oil, peppermint oil, cool mint oil, spearmint oil, perilla oil, star anise oil, cinnamon oil, rose oil, camphor oil, etc. Thickening agent: for example, cellulose-based polymer compounds such as methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, sodium carboxymethyl cellulose, etc.; polyvinyl-based polymer compounds such as polyvinyl pyrrolidone, polyvinyl alcohol, etc.; carboxyvinyl polymer; guar gum; hydroxypropyl guar gum; gum arabic; karaya gum; xanthan gum; agar; alginic acid and its salts (sodium salt, etc.); mucopolysaccharides such as heparin-like substances, heparin, heparin sulfate, heparan sulfate, heparinoid, hyaluronic acid and its salts (sodium salt, etc.), chondroitin sulfate and its salts (sodium salt, etc.); starch; chitin and its derivatives; chitosan and its derivatives; carrageenan; monosaccharides such as glucose, etc. Buffer: for example, boric acid buffer, phosphate buffer, carbonate buffer, acetate buffer, lactate buffer, succinate buffer, citrate buffer, Tris buffer, AMPD buffer, etc. Stabilizer: for example, edetic acid, edetic acid salts (disodium edetate, calcium disodium edetate, trisodium edetate, tetrasodium edetate), sodium formaldehyde sulfoxylate (rongalit), aluminum monostearate, glycerin monostearate, cyclodextrin, monoethanolamine, dibutylhydroxytoluene, etc. Preservatives: For example, alkyl polyaminoethyl glycine quaternary ammonium salts (e.g., benzalkonium chloride, benzethonium chloride, etc.), chlorhexidine gluconate, poly(diallyldimethylammonium chloride), zinc chloride, sodium benzoate, ethanol, chlorobutanol, sorbic acid, potassium sorbate, sodium dehydroacetate, methyl paraben, ethyl paraben, propyl paraben, butyl paraben, oxyquinoline sulfate, phenethyl alcohol, benzyl alcohol, biguanide compounds (specifically, polyhexanide hydrochloride (polyhexamethylene biguanide), alexidine, etc.), glochil (trade name, manufactured by Rhodia), etc. Isotonic agents: For example, potassium chloride, calcium chloride, sodium chloride, magnesium chloride, potassium acetate, sodium acetate, sodium hydrogen carbonate, sodium carbonate, sodium thiosulfate, magnesium sulfate, glycerin, propylene glycol, etc. Sugar alcohols: For example, xylitol, sorbitol, mannitol, etc. These may be in the form of d-form, l-form, or dl-form. Oils: For example, vegetable oils such as sesame oil, castor oil, soybean oil, olive oil; animal oils such as squalane; mineral oils such as liquid paraffin, petrolatum, etc.

[0063] When the ophthalmic composition according to this embodiment contains water, from the viewpoint of more significantly achieving the effects of the present invention, for example, based on the total amount of the ophthalmic composition, the water content is preferably 80 w / v% or more and less than 100 w / v%, more preferably 85 w / v% or more and 99.5 w / v% or less, and still more preferably 90 w / v% or more and 99.2 w / v% or less.

[0064] The water used in the ophthalmic composition according to this embodiment may be pharmaceutically, pharmacologically (pharmaceutically) or physiologically acceptable. Examples of such water include distilled water, tap water, purified water, sterilized purified water, water for injection, and distilled water for injection, etc. Their definitions are based on the seventeenth revised Japanese Pharmacopoeia.

[0065] The ophthalmic composition according to this embodiment can be prepared by adding and mixing a desired amount of component (A) (or component (A')) and, if necessary, other components to a desired concentration. For example, these components can be dissolved or dispersed in purified water, adjusted to a predetermined pH and osmotic pressure, and sterilized by filtration sterilization or the like.

[0066] The ophthalmic composition according to this embodiment can take various dosage forms according to the purpose. Examples of dosage forms include, for example, liquid preparations, gel preparations, semi-solid preparations (such as ointments), and the like.

[0067] The ophthalmic composition according to this embodiment can be used, for example, as eye drops (also referred to as eye drops or eye medications. Eye drops include eye drops that can be instilled during contact lens wear.), artificial tears, eye washes (also referred to as eye wash solutions or eye wash medications. Eye washes include eye washes that can be used during contact lens wear.), contact lens compositions [such as contact lens soaking solutions, contact lens care compositions (contact lens disinfectants, contact lens preservatives, contact lens cleaners, contact lens cleaning and preservative solutions), etc.]. Note that "contact lens" includes hard contact lenses and soft contact lenses (including both ionic and non-ionic types, and including both silicone hydrogel contact lenses and non-silicone hydrogel contact lenses).

[0068] Since the ophthalmic composition according to this embodiment can exhibit the effects of the present invention more significantly, it is preferably an eye drop (including eye drops that can be instilled during contact lens wear). When the ophthalmic composition according to this embodiment is an eye drop, its usage and dosage are not particularly limited as long as they are effective and have few side effects. For example, in the case of adults (15 years old and above) and children 7 years old and above, a method of instilling 1 to 3 drops, 1 to 2 drops, or 2 to 3 drops 5 to 6 times a day can be exemplified.

[0069] The ophthalmic composition according to this embodiment is provided by being contained in an arbitrary container. The container for containing the ophthalmic composition according to this embodiment is not particularly limited. For example, it may be made of glass or may be made of plastic. Preferably, it is made of plastic. Examples of the plastic include polyethylene terephthalate, polyarylate, polyethylene naphthalate, polycarbonate, polyethylene, polypropylene, polyimide, copolymers of the monomers constituting these, and mixtures of two or more of these. Preferably, it is polyethylene terephthalate. Further, the container for containing the ophthalmic composition according to this embodiment may be a transparent container through which the inside of the container can be visually recognized, or may be an opaque container in which it is difficult to visually recognize the inside of the container. Preferably, it is a transparent container. Here, the "transparent container" includes both a colorless transparent container and a colored transparent container.

[0070] A nozzle may be attached to the container for containing the ophthalmic composition according to this embodiment. The material of the nozzle is not particularly limited. For example, it may be made of glass or may be made of plastic. Preferably, it is made of plastic. Examples of the plastic include polybutylene terephthalate, polyethylene, polypropylene, polyethylene terephthalate, polyethylene naphthalate, copolymers of the monomers constituting these, and mixtures of two or more of these. From the viewpoint of further enhancing the effects of the present invention, polypropylene, polyethylene, polyethylene terephthalate, polybutylene terephthalate, and polyethylene naphthalate are preferable as the material of the nozzle, and polyethylene is more preferable.

[0071] The container for containing the ophthalmic composition according to this embodiment may be a multi-dose type that contains a plurality of usage amounts, or may be a unit-dose type that contains a single usage amount.

[0072] The ophthalmic composition according to this embodiment is preferably filled in a container having an internal volume of 4 to 30 mL, more preferably filled in a container having an internal volume of 5 to 20 mL, still more preferably filled in a container having an internal volume of 6 to 16 mL, and even more preferably filled in a container having an internal volume of 10 to 15 mL.

[0073] The ophthalmic composition according to this embodiment can protect corneal epithelial stem cells from damage by light by containing at least one selected from the group consisting of sulfites, vitamin Es, and zinc sulfate. Further, the ophthalmic composition according to this embodiment can protect corneal epithelial stem cells from damage by light by containing at least one selected from the group consisting of sulfites, vitamin Es, aminoethylsulfonic acid and its salts, flavin adenine dinucleotide and its salts, pyridoxine and its salts, neostigmine and its salts, naphazoline and its salts, aspartic acid and its salts, zinc sulfate, and vitamin As. Here, since the corneal epithelial stem cells are protected from damage by light, the corneal epithelial cells and the corneal tissue composed of the corneal epithelial cells are regenerated. Therefore, the ophthalmic composition according to this embodiment can also be suitably used as a corneal tissue regenerating agent from damage by light. Further, since the corneal epithelial stem cells are protected from damage by light, the corneal epithelial cells and the corneal tissue composed of the corneal epithelial cells are recovered. Therefore, the ophthalmic composition according to this embodiment can also be suitably used as a corneal tissue recovering agent from damage by light. Furthermore, since the corneal epithelial stem cells are protected from damage by light, the corneal epithelial cells and the corneal tissue composed of the corneal epithelial cells are repaired. Therefore, the ophthalmic composition according to this embodiment can also be suitably used as a corneal tissue repairing agent from damage by light.

[0074] 〔2. Method for evaluating the protective effect of stem cells from damage by blue light〕 A method for evaluating the protective effect of blue light on stem cells according to this embodiment (simply referred to as "the evaluation method according to this embodiment") includes a step of irradiating stem cells with blue light (simply referred to as "the irradiation step"), and a step of measuring the cell viability of the stem cells (simply referred to as "the measurement step").

[0075] In the irradiation step, the stem cells are irradiated with blue light for a predetermined time.

[0076] As the blue light for irradiating the stem cells, blue light having a wavelength in the range of 380 to 500 nm can be used. Also, the irradiation time of the blue light on the stem cells can be appropriately set, for example, in the range of 1 to 15 hours.

[0077] The stem cells are not particularly limited as long as they are stem cells that may be damaged by irradiation with blue light, and examples include corneal epithelial stem cells.

[0078] In the measurement step, the cell viability of the stem cells in the absence and presence of the test substance is measured.

[0079] In this specification, "cell viability" means the ratio (%) of the number of live cells measured in a test solution with or without the test substance after blue light irradiation, with the number of live cells in the control being 100%. Here, the control means a test solution that is not irradiated with blue light and does not contain the test substance. When corneal epithelial stem cells are used as the stem cells, the measurement of the number of live cells can be performed by irradiating each test solution with blue light, adding a cell number measurement reagent, culturing under predetermined conditions, and measuring the absorbance at a predetermined wavelength.

[0080] In the evaluation method according to this embodiment, the measurement step may further include a step of calculating a cell protection rate based on the cell viability measured above. Here, "cell protection rate" means a numerical value calculated by the following formula. (Formula) Cell protection rate (%) = { (Cell viability in the presence of the test substance - Cell viability in the absence of the test substance) / Cell viability in the absence of the test substance} × 100

[0081] The evaluation method according to this embodiment preferably includes a step of evaluating the protective effect of stem cells from damage by blue light (simply referred to as the "evaluation step") based on the change in the cell viability of stem cells obtained in the above measurement step.

[0082] In the evaluation step, the protective effect of stem cells from damage by blue light is evaluated based on the change in the cell viability of stem cells in the absence and presence of the test substance obtained in the above measurement step. Specifically, when the cell viability in the presence of the test substance is increased compared to the cell viability in the absence of the test substance, it can be evaluated that the test substance has a protective effect on stem cells from damage by blue light.

[0083] In the evaluation method according to this embodiment, when the measurement step includes a step of calculating the cell protection rate, in the evaluation step, the protective effect of stem cells from damage by blue light is evaluated based on the calculated cell protection rate. Specifically, when the cell protection rate is a positive value, it can be evaluated that the test substance has a protective effect on stem cells from damage by blue light.

Example

[0084] Hereinafter, the present invention will be specifically described based on test examples, but the present invention is not limited thereto.

[0085] 〔Test Example 1: Cell protection rate of mouse corneal epithelial stem cells by blue light irradiation (1)〕 With the compositions shown in Table 1, each aqueous composition was prepared according to a conventional method and used as a test solution. The unit of each component in Table 1 is w / v%.

[0086] Mouse corneal epithelial stem cells (TKE2) were seeded onto a culture plate (12-well, manufactured by Corning Japan) and cultured under conditions of 37°C, 5% CO2, and 90% humidity until confluent. As the culture medium, KSFM (keratinocyte serum-free medium (containing Bovine Pituitary Extract and EGF), manufactured by Thermo Fisher Scientific) was used.

[0087] The culture medium was aspirated and removed from each well, and 1 mL of each test solution was added to each well, followed by incubation at 37°C and 5% CO2 for 1 hour. Next, after irradiating with blue light (wavelength 470 nm) for 14 hours under conditions of 37°C and 5% CO2, the number of viable cells in each well was measured.

[0088] To measure the number of viable cells, the cell counting reagent Cell Counting Kit-8 (manufactured by DOJINDO) was added to each well, cultured at 37°C and 5% CO2 for 1 hour, and then the absorbance at 450 nm was measured. The ratio of the number of viable cells measured for each test solution to the number of viable cells in the control, with the number of viable cells in the control set as 100%, was defined as the cell survival rate (%). Note that the control was treated in the same manner as Comparative Example 1-1 except that it was not irradiated with blue light.

[0089] Next, the cell protection rate (%) of each example was determined according to the following (Formula 1). (Formula 1) Cell protection rate (%) = { (Cell survival rate of the example - Cell survival rate of Comparative Example 1-1) / Cell survival rate of Comparative Example 1-1} × 100 The results are shown in Table 1.

[0090]

Table 1

[0091] It was confirmed that the corneal epithelial stem cells were protected from damage by blue light by adding sodium pyrosulfite.

[0092] [Test Example 2: Cell protection rate of mouse corneal epithelial stem cells by blue light irradiation (2)] Using the composition shown in Table 2, each aqueous composition was prepared according to a conventional method and used as a test solution. The unit of each component in Table 2 is w / v%.

[0093] Mouse corneal epithelial stem cells (TKE2) were seeded on a culture plate (12-well, manufactured by Corning Japan) and cultured under the conditions of 37 °C, 5% CO2, and 90% humidity until confluent. As the culture medium, KSFM (keratinocyte serum-free medium (containing Bovine Pituitary Extract, EGF), manufactured by Thermo Fisher Scientific) medium was used.

[0094] The culture medium was aspirated and removed from each well, 1 mL of each test solution was added to each well, and the mixture was incubated at 37 °C and 5% CO2 for 1 hour. Next, after irradiating with blue light (wavelength 470 nm) at 37 °C and 5% CO2 for 14 hours, the number of viable cells in each well was measured.

[0095] The measurement of the number of viable cells was performed by adding the cell number measurement reagent Cell Counting Kit-8 (manufactured by DOJINDO) to each well, culturing at 37 °C and 5% CO2 for 1 hour, and then measuring the absorbance at 450 nm. The ratio of the number of viable cells measured for each test solution when the number of viable cells in the control was set to 100% was defined as the cell survival rate (%). The control was treated in the same manner as in Comparative Example 2-1 except that it was not irradiated with blue light.

[0096] Next, the cell protection rate (%) of each example was determined according to the following (Formula 2). (Formula 2) Cell protection rate (%) = { (Cell survival rate of the example - Cell survival rate of Comparative Example 2-1) / Cell survival rate of Comparative Example 2-1} × 100 The results are shown in Table 2.

[0097]

Table 2

[0098] It was confirmed that adding tocopherol acetate protected corneal epithelial stem cells from damage caused by blue light.

[0099] 〔Test Example 3: Cell protection rate of mouse corneal epithelial stem cells by blue light irradiation (3)〕 Using the compositions shown in Table 3, each aqueous composition was prepared according to a conventional method and used as a test solution. The unit of each component in Table 3 is w / v%.

[0100] Mouse corneal epithelial stem cells (TKE2) were seeded on a culture plate (12 well, manufactured by Corning Japan), and cultured until confluent under the conditions of 37 °C, 5% CO2, and 90% humidity. As the culture medium, KSFM (keratinocyte serum-free medium (containing bovine pituitary extract, EGF), manufactured by Thermo Fisher Scientific) medium was used.

[0101] The culture medium was aspirated and removed from each well, 1 mL of each test solution was added to each well, and incubated at 37 °C and 5% CO2 for 6 hours. Next, after irradiating with blue light (wavelength 470 nm) at 37 °C and 5% CO2 for 14 hours, the number of viable cells in each well was measured.

[0102] The measurement of the number of viable cells was performed by adding the cell number measurement reagent Cell Counting Kit-8 (manufactured by DOJINDO) to each well, culturing at 37 °C and 5% CO2 for 1 hour, and then measuring the absorbance at 450 nm. The ratio of the number of viable cells measured for each test solution when the number of viable cells in the control was set to 100% was defined as the cell survival rate (%). The control was treated in the same manner as Comparative Example 3-1 except that it was not irradiated with blue light.

[0103] Next, the cell protection rate (%) of each example was determined according to the following (Formula 3). (Formula 3) Cell protection rate (%) = {(Cell viability in the example - Cell viability in Comparative Example 3-1) / Cell viability in Comparative Example 3-1} × 100 The results are shown in Table 3.

[0104]

Table 3

[0105] It was confirmed that corneal epithelial stem cells were protected from damage by blue light by adding zinc sulfate.

[0106] 〔Formulation Example〕 Eye drops or eye drops for soft contact lenses (Formulation Examples 1 to 16) are prepared by a conventional method according to the formulations described in Table 4 and Table 5 below. In addition, the unit of each component amount in Table 4 and Table 5 below is w / v% unless otherwise specified in the table. Also, in Table 4 and Table 5, retinol palmitate indicates the amount of formulation per 100 mL.

[0107]

Table 4

[0108]

Table 5

Claims

1. An ophthalmic composition for protecting corneal epithelial cells from blue light damage, containing at least one selected from the group consisting of salts of pyrosulfurous acid and zinc sulfate (excluding those containing α-tocopheryl retinoate).

2. The ophthalmic composition according to Claim 1, wherein the corneal epithelial cells are corneal epithelial stem cells.

3. A corneal tissue regenerating agent from blue light damage, containing at least one selected from the group consisting of salts of pyrosulfurous acid and zinc sulfate (excluding those containing α-tocopheryl retinoate).

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