JP2020075918A - Ophthalmic composition for promoting corneal epithelium wound cure - Google Patents

Abstract

To provide an ophthalmic composition used for promoting curing of a corneal epithelium wound.SOLUTION: An ophthalmic composition with an osmotic pressure of 200-260 mOsm has the action of promoting curing of a corneal epithelium wound. An eye suffering from a corneal epithelium wound is treated with the ophthalmic composition, to promote curing of the corneal epithelium wound.SELECTED DRAWING: None

Description

  The present invention relates to ophthalmic compositions used for promoting healing of corneal epithelial wounds.

  The cornea is composed of corneal epithelium, Bowman's membrane, corneal stroma, Descemet's membrane, and corneal endothelium. Among these, the corneal epithelium located at the outermost layer is a tissue in which a wound is likely to occur due to internal factors or external factors. Normally, even if a wound occurs in the corneal epithelium, if the epithelial stem cells are healthy, they are naturally healed by the self-repairing function. Wound healing of corneal epithelium proceeds through three phases of spreading and migration of epithelial cells (phase 1), proliferation of epithelial cells (phase 2), and differentiation of epithelial cells (phase 3).

  As described above, the corneal epithelial wound is naturally healed by the self-repairing function, but it may take a long time or the wound closure may be insufficient. During the healing period of corneal epithelial wounds or when the wound closure is insufficient, the barrier function of the corneal epithelium cannot be fully exerted, the homeostasis of the cornea becomes insufficient, and the risk of bacterial infection increases. .. Therefore, promoting healing of corneal epithelial wounds has become important in preventing eye diseases and maintaining the eye in a healthy state.

  Heretofore, various therapeutic agents effective for treating corneal epithelial wounds have been studied (for example, Patent Documents 1 to 3).

Japanese Patent Publication No. 2005-529906 Japanese Patent Publication No. 2013-523829 JP-A-2-212433

  An object of the present invention is to provide an ophthalmic composition used for promoting healing of corneal epithelial wounds.

  The present inventors have found that an ophthalmic composition having an osmotic pressure of 200 to 260 mOsm has an action of promoting healing of a corneal epithelial wound, and applies the ophthalmic composition to an eye having a corneal epithelial wound. Have been found to promote healing of corneal epithelial wounds. The present invention has been completed by further studies based on such findings.

As one aspect of the present invention, the following ophthalmic composition is provided.
Item 1-1. An ophthalmic composition for promoting corneal epithelial wound healing, which has an osmotic pressure of 200 to 260 mOsm.
Item 1-2. The ophthalmic composition for promoting corneal epithelial wound healing according to Item 1-1, which has an osmotic pressure of 220 to 260 mOsm.
Item 1-3. The ophthalmic composition for promoting corneal epithelial wound healing according to Item 1-1 or 1-2, which comprises an alkali metal salt and / or a polyhydric alcohol.
Item 1-4. Item 3. The ophthalmic composition for promoting corneal epithelial wound healing according to Item 1-3, wherein the alkali metal salt is sodium chloride.
Item 1-5. Item 3. The ophthalmic composition for promoting corneal epithelial wound healing according to Item 1-3, wherein the polyhydric alcohol is propylene glycol and / or glycerin.
Item 1-6. The ophthalmic composition for promoting corneal epithelial wound healing according to Item 1-1, which has an osmotic pressure of 210 to 255 mOsm and contains sodium chloride.
Item 1-7. The ophthalmic composition for promoting corneal epithelial wound healing according to Item 1-1, which has an osmotic pressure of 220 to 258 mOsm and contains glycerin.
Item 1-8. The ophthalmic composition for promoting corneal epithelial wound healing according to Item 1-1, which has an osmotic pressure of 220 to 255 mOsm and contains propylene glycol.
Item 1-9. The ophthalmic composition for promoting corneal epithelial wound healing according to Item 1-1, which has an osmotic pressure of 219 to 255 mOsm and contains glycerin, boric acid, and borax.
Item 1-10. The ophthalmic composition for promoting corneal epithelial wound healing according to any one of Items 1-1 to 1-4, which comprises sodium chloride and tromethamol.
Item 1-11. The ophthalmic composition for promoting corneal epithelial wound healing according to any one of Items 1-1 to 1-4, comprising at least one selected from the group consisting of edetic acid and its salt, and 1-menthol.
Item 1-12. The corneal epithelial wound healing according to any one of Items 1-1 to 1-4, comprising at least one selected from the group consisting of pyridoxine, chlorpheniramine, cyanocobalamin, aspartic acid, taurine, neostigmine, and salts thereof. Ophthalmic composition for promotion.
Item 1-13. The ophthalmic composition for promoting corneal epithelial wound healing according to any one of Items 1-1 to 1-12, which is an eye drop.

In addition, as another aspect of the present invention, the following method is provided.
Item 2-1. A method for imparting a corneal epithelial wound healing promoting action to an ophthalmic composition, which comprises the step of adjusting the osmotic pressure of the ophthalmic composition to 200 to 260 mOsm.
Item 2-2. Item 2. The method according to Item 2-1, wherein the osmotic pressure is 220 to 260 mOsm.
Item 2-3. Item 3. The method according to Item 2-1 or 2-2, wherein an alkali metal chloride and / or a polyhydric alcohol is added to the ophthalmic composition.
Item 2-4. Item 3. The method according to Item 2-3, wherein the chloride of the alkali metal is sodium chloride.
Item 2-5. Item 3. The method according to Item 2-3, wherein the polyhydric alcohol is propylene glycol and / or glycerin.
Item 2-6. Item 2. The method according to Item 2-1, wherein the osmotic pressure is 210 to 255 mOsm, and sodium chloride is added.
Item 2-7. Item 2. The method according to Item 2-1, wherein the osmotic pressure is 220 to 258 mOsm, and glycerin is added.
Item 2-8. The method according to Item 2-1, wherein the osmotic pressure is 220 to 255 mOsm, and propylene glycol is blended.
Item 2-9. The method according to Item 2-1, wherein the osmotic pressure is 219 to 255 mOsm, and glycerin, boric acid, and borax are blended.
Item 2-10. Item 5. The method according to any one of Items 2-1 to 2-4, wherein sodium chloride and tromethamol are added to the ophthalmic composition.
Item 2-11. Item 5. The method according to any one of Items 2-1 to 2-4, wherein the ophthalmic composition contains at least one selected from the group consisting of edetic acid and a salt thereof, and 1-menthol.
Item 2-12. In the ophthalmic composition, at least one selected from the group consisting of pyridoxine, chlorpheniramine, cyanocobalamin, aspartic acid, taurine, neostigmine, and salts thereof is blended, and the items 2-1 to 2-4. The method described in either.
Item 2-13. Item 13. The method according to any one of Items 2-1 to 2-12, wherein the ophthalmic composition is an eye drop.

  According to the ophthalmic composition of the present invention, the low osmotic pressure of 200 to 260 mOsm can effectively promote the healing of corneal epithelial wounds. In addition, the ophthalmic composition of the present invention can promote healing of corneal epithelial wounds without using a pharmacological component, and thus can be a highly safe eye drop that replaces hyaluronic acid eye drops and the like. Furthermore, the ophthalmic composition of the present invention can reduce corneal epithelial disorders such as toxic keratopathy, which is a concern with a therapeutic agent for glaucoma.

FIG. 3 is a diagram showing the results of performing a scratch assay under various osmotic pressure conditions using immortalized human corneal epithelial cells in Test Example 1.

1. Definitions As used herein, the term “corneal epithelial wound” refers to a state in which corneal epithelial tissue or corneal epithelial cells are deficient due to mechanical, chemical or biological stimulation. “Corneal epithelial wound” includes, for example, corneal epithelial defect associated with an endogenous disease such as Sjogren's syndrome, Stevens-Johnson syndrome, corneal ulcer, meibomian adenitis, allergic conjunctivitis, dry eye; postoperative, drug-induced, It includes corneal epithelial defects associated with extrinsic diseases such as trauma, chemical trauma, burns, and contact lens wearing; corneal epithelial defects associated with ocular allergic diseases associated with corneal lesions such as spring catarrhal and atopic keratoconjunctivitis.

  In the present specification, “healing of corneal epithelial wound” means that the corneal epithelial tissue or corneal epithelial cells spread, migrate (migrate), proliferate or differentiate at the site where the corneal epithelium is defective, and the defective site is closed. A biological reaction that goes toward a state.

  In the present specification, “for healing promotion” refers to a substance that promotes a biological reaction toward a state in which a site where the corneal epithelium is defective is blocked. If the site where the corneal epithelium is lost is blocked, it is suitable for healing.

  In the present specification, “providing a healing-promoting action” refers to having a function of promoting a biological reaction toward a closed state at a site where the corneal epithelium is defective.

  As used herein, the "ophthalmic composition" is a pharmaceutical composition for ophthalmic use.

  In the present specification, the "osmotic pressure" is a value measured according to the method defined in "30. Osmotic pressure measuring method (osmolarity measuring method)" of "General test method" of the 17th revised Japanese Pharmacopoeia Is.

2. Description of the Preferred Embodiments The following is a description of the preferred embodiments, it being understood that this embodiment is illustrative of the invention and the scope of the invention is not limited to such preferred embodiments. .. It should be understood that those skilled in the art can easily make modifications, changes and the like within the scope of the present invention with reference to the preferred embodiments described later. Those skilled in the art can appropriately combine any of these embodiments.

3. Ophthalmic Composition Conventionally, as an ophthalmic composition that promotes healing of corneal epithelial wounds, an eye drop containing sodium hyaluronate as an active ingredient, a composition containing xanthan gum as an active ingredient, a composition containing ketorolac and carboxymethylcellulose, and vitronectin. Although eye drops and the like containing as an active ingredient are known, the relationship between healing of corneal epithelial wounds and osmotic pressure of an ophthalmic composition is not known. Under such circumstances, the present inventors have found that a low osmotic ophthalmic composition can promote healing of corneal epithelial wounds. Specifically, it has been found that an ophthalmic composition having an osmotic pressure of 200 to 260 mOsm can effectively promote healing of corneal epithelial wounds.

  That is, in one aspect, the present invention provides an ophthalmic composition for promoting corneal epithelial wound healing, which has an osmotic pressure of 200 to 260 mOsm.

[Osmotic pressure]
The osmotic pressure in the ophthalmic composition of the present invention is 200 to 260 mOsm. By adjusting to such an osmotic pressure, it becomes possible to effectively promote healing of a corneal epithelial wound. However, if the osmotic pressure is too low, discomfort tends to appear when instilled. While suppressing the discomfort at the time of instillation due to such a low osmotic pressure, from the viewpoint of effectively exerting a healing promoting effect on the corneal epithelial wound, the osmotic pressure of the ophthalmic composition of the present invention is preferably 218 to 260 mOsm, more preferably 220 to 260 mOsm, still more preferably 240 to 260 mOsm.

  In order to adjust the osmotic pressure to the range described above, the kind and concentration of the components to be blended in the ophthalmic composition of the present invention may be adjusted, and the osmotic pressure of the ophthalmic composition can be appropriately set by those skilled in the art. It is a matter.

[Ingredients]
The ophthalmic composition of the present invention contains water as a base and may contain various components in order to satisfy the above-mentioned osmotic pressure.

  The ophthalmic composition of the present invention preferably contains an osmotic pressure adjusting agent and / or a buffering agent in order to satisfy the above-mentioned osmotic pressure.

  The osmotic pressure adjusting agent is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include alkali metal salts, alkaline earth metal salts, polyhydric alcohols and sugars. Specific examples of the alkali metal salt include chlorides of alkali metals such as sodium chloride and potassium chloride; alkali metal salts of organic acids such as sodium acetate and potassium acetate; borax, sodium hydrogen sulfite, sodium hydrogen carbonate, sodium carbonate. Inorganic acid alkali metal salts such as disodium hydrogen phosphate, sodium dihydrogen phosphate, and the like. Specific examples of the alkaline earth metal salt include chlorides of alkaline earth metals such as calcium chloride and magnesium chloride. Specific examples of the polyhydric alcohol include glycerin, propylene glycol, butylene glycol, polyethylene glycol and the like. Specific examples of the saccharide include sorbitol, glucose, mannitol and the like. These osmotic pressure adjusting agents may be in the form of solvates such as hydrates. For example, in the case of disodium hydrogen phosphate, it may be disodium hydrogen phosphate dodecahydrate.

  Among these osmotic pressure regulators, preferably alkali metal salts, and polyhydric alcohols, more preferably alkali metal chlorides, inorganic acid alkali metal salts, and polyhydric alcohols, particularly preferably sodium chloride, hydrogen phosphate. Examples include disodium, glycerin, and propylene glycol.

  These osmotic pressure regulators may be used alone or in combination of two or more.

  When the osmotic pressure adjusting agent is contained in the ophthalmic composition of the present invention, the concentration thereof satisfies the above-mentioned osmotic pressure depending on the kind of the osmotic pressure adjusting agent to be used, the kind and the concentration of other components to be blended, and the like. The amount may be appropriately set so that it is 0.001 to 5 w / v%, and preferably 0.005 to 3 w / v%.

  The buffering agent plays a role of adjusting the osmotic pressure and imparting a buffering effect. The buffer used in the present invention is not particularly limited as long as it is pharmaceutically acceptable. For example, phosphate buffer, citrate buffer, borate buffer, Tris buffer, tartaric acid buffer. , Acetate buffers, amino acid buffers and the like.

  Specific examples of the phosphate buffer include phosphoric acid and / or a salt thereof. The salt of phosphoric acid is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include dialkali metal hydrogenphosphate salts such as disodium hydrogenphosphate, dipotassium hydrogenphosphate; sodium dihydrogenphosphate. , Alkali dihydrogen phosphate salts such as potassium dihydrogen phosphate; trialkali metal phosphate salts such as trisodium phosphate, tripotassium phosphate and the like. The salt of phosphoric acid may be in the form of a solvate such as a hydrate. For example, in the case of disodium hydrogen phosphate, the form of dodecahydrate or sodium dihydrogen phosphate. In some cases, it may be in the form of dihydrate. As the phosphate buffer, one type may be selected from phosphoric acid and salts thereof and used alone, or two or more types may be used in combination. Among phosphoric acid and its salts, preferably phosphate, more preferably at least one of dialkali metal hydrogenphosphate and alkali metal dihydrogenphosphate, particularly preferably disodium hydrogenphosphate and dihydrogenphosphate. At least one of sodium may be mentioned.

  Specific examples of the citric acid buffer include citric acid and / or a salt thereof. The citric acid salt is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt. Be done. Also, the salt of citric acid may be in the form of a solvate such as a hydrate. As the citric acid buffer, one kind may be selected from citric acid and salts thereof and used alone, or two or more kinds may be used in combination.

  Specific examples of the borate buffer include boric acid and / or salts thereof. Boric acid is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include orthoboric acid, metaboric acid, tetraboric acid and the like. Among these boric acids, orthoboric acid and tetraboric acid are preferable. These boric acids may be used alone or in combination of two or more. The salt of boric acid is not particularly limited as long as it is pharmaceutically acceptable, but is an alkali metal salt such as sodium salt and potassium salt; alkaline earth metal salt such as calcium salt and magnesium salt; aluminum salt; Examples thereof include organic amine salts such as triethylamine, triethanolamine, morpholine, piperazine and pyrrolidine. The boric acid / or its salt may also be in the form of a hydrate, such as borax. As the boric acid buffer, one kind may be selected from boric acid and salts thereof and used alone, or two or more kinds may be used in combination.

  Specific examples of the Tris buffer include trometamol and / or a salt thereof. The salt of trometamol is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include organic acid salts such as acetate; and organic acid salts such as hydrochloride and sulfonate. As the tris acid buffer, one kind may be selected from trometamol and a salt thereof and used alone, or two or more kinds may be used in combination.

  Specific examples of the tartaric acid buffer include tartaric acid and / or its salt. The tartaric acid salt is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include alkali metal salts such as sodium salts and potassium salts; alkaline earth metal salts such as calcium salts and magnesium salts. .. Further, the salt of tartaric acid may be in the form of a solvate such as a hydrate. As the tartaric acid buffer, one kind may be selected from tartaric acid and salts thereof and used alone, or two or more kinds may be used in combination.

  Specific examples of the acetic acid buffer include acetic acid and / or a salt thereof. The acetic acid salt is not particularly limited as long as it is pharmaceutically acceptable, but examples thereof include alkali metal salts such as sodium salts and potassium salts; alkaline earth metal salts such as calcium salts and magnesium salts; ammonium salts and the like. Is mentioned. Further, the salt of acetic acid may be in the form of a solvate such as a hydrate. As the acetic acid buffer, one kind may be selected from acetic acid and salts thereof and used alone, or two or more kinds may be used in combination.

  Specific examples of the amino acid buffer include acidic amino acids, neutral amino acids, and salts thereof. Specific examples of the acidic amino acid include aspartic acid and glutamic acid. The salt of the acidic amino acid is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include alkali metal salts such as sodium salt and potassium salt. Specific examples of the neutral amino acid include glycine, alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, proline, serine, threonine, cysteine, methionine, asparagine, glutamine, and taurine. The neutral amino acid salt is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include alkali metal salts such as sodium salt and potassium salt. As the amino acid buffer, one kind may be selected from acidic amino acids and salts thereof and used alone, or two or more kinds may be used in combination.

  These buffers may be used alone or in combination of two or more.

  Among these buffers, preferred are phosphate buffers, citrate buffers, borate buffers, and Tris buffers.

  When a buffer is contained in the ophthalmic composition of the present invention, the concentration thereof is appropriately selected so as to satisfy the above-mentioned osmotic pressure depending on the type of the buffer used, the type and concentration of other components to be blended, and the like. It may be set, but for example, 0.001 to 5 w / v%, preferably 0.005 to 3.5 w / v%, and more preferably 0.01 to 2 w / v% can be mentioned.

  As a specific example of an ophthalmic composition containing the osmotic pressure adjusting agent and / or buffer, the osmotic pressure satisfying the above range, sodium chloride is contained, and the osmotic pressure is 200 to 260 mOsm, preferably 210 to 255 mOsm. An ophthalmic composition containing glycerin and having an osmotic pressure of 200 to 260 mOsm, preferably 220 to 258 mOsm; containing propylene glycol, and having an osmotic pressure of 200 to 260 mOsm, preferably 220 to 255 mOsm Ophthalmic composition; containing glycerin, boric acid and borax, and having an osmotic pressure of 200 to 260 mOsm, preferably 219 to 255 mOsm; containing sodium chloride and tromethamol, and having an osmotic pressure of 200 to Examples thereof include an ophthalmic composition having 260 mOsm, preferably 221 mOsm.

  In the ophthalmic composition of the present invention, in addition to the above components, a surfactant, a thickener, a solubilizing agent, a chelating agent, and a cooling agent may be added as long as the above-mentioned osmotic pressure is satisfied. Additives such as agents, stabilizers, pH adjusters and preservatives may be contained.

  The surfactant is not particularly limited as long as it is pharmaceutically acceptable, for example, tyloxapol, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene block copolymer, polyoxyethylene sorbitan fatty acid ester, octoxy. Nonionic surfactants such as nols; amphoteric surfactants such as alkyldiaminoethylglycine, lauryldimethylaminoacetic acid betaine; alkyl sulfates, N-acyl taurine salts, polyoxyethylene alkyl ether phosphates, polyoxyethylene alkyls Anionic surfactants such as ether sulfates; cationic surfactants such as alkylpyridinium salts and alkylamine salts. These surfactants may be used alone or in combination of two or more.

  The thickener is not particularly limited as long as it is pharmaceutically acceptable, for example, carboxyvinyl polymer, polyvinylpyrrolidone, polyethylene glycol, polyvinyl alcohol, xanthan gum, sodium chondroitin sulfate, sodium hyaluronate, gellan gum, alginic acid, Water-soluble polymers such as polycarbophil; celluloses such as hydroxyethyl cellulose, methyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, and the like. These thickeners may be used alone or in combination of two or more.

  The solubilizer is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include polyoxyethylene castor oil, ethanol, polyethylene glycol monostearate, polyoxyl stearate, castor oil, carbomer copolymer type A and the like. Can be mentioned. These solubilizers may be used alone or in combination of two or more.

  The chelating agent is not particularly limited as long as it is pharmaceutically acceptable, for example, edetic acid, citric acid, succinic acid, ascorbic acid, trihydroxymethylaminomethane, nitrilotriacetic acid, 1-hydroxyethane-1, 1-diphosphonic acid, polyphosphoric acid, metaphosphoric acid, hexametaphosphoric acid, these salts, etc. are mentioned. The form of the salt is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include alkali metal salts such as sodium salt and potassium salt. Also, these chelating agents may be in the form of hydrates. These chelating agents may be used alone or in combination of two or more.

  The cooling agent is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include l-menthol, borneol, camphor, eucalyptus oil, geraniol, bergamot oil and the like. These cooling agents may be used alone or in combination of two or more.

  The stabilizer is not particularly limited as long as it is pharmaceutically acceptable, for example, polyvinylpyrrolidone, sulfite, monoethanolamine, cyclodextrin, dextran, ascorbic acid, taurine, tocopherol, dibutylhydroxytoluene, thiol. Examples thereof include sodium sulfate and sodium hydrogen sulfite. These stabilizers may be used alone or in combination of two or more.

  The pH adjuster is not particularly limited as long as it is pharmaceutically acceptable, but examples thereof include acids such as hydrochloric acid, acetic acid, boric acid, aminoethyl sulfonic acid, epsilon aminocaproic acid; sodium hydroxide, potassium hydroxide, Examples include alkali such as borax, triethanolamine, monoethanolamine, sodium hydrogen carbonate and sodium carbonate. These pH adjusters may be used alone or in combination of two or more.

  The concentrations of these additives may be appropriately set within a range that satisfies the above-mentioned osmotic pressure, depending on the type of the additive used, the properties to be imparted to the aqueous liquid agent, and the like.

  Among these additives, suitable examples include at least one selected from the group consisting of edetic acid and its salt, and 1-menthol.

  When the ophthalmic composition of the present invention contains edetic acid and / or a salt thereof, the concentration thereof is not particularly limited, but is, for example, 0.001 to 0.127 w / v%, preferably 0.005 to 0. 0.1 w / v%, and more preferably 0.01 to 0.05 w / v%.

  When l-menthol is contained in the ophthalmic composition of the present invention, its concentration is not particularly limited, but for example, 0.0001 to 0.5 w / v%, preferably 0.0005 to 0.1 w. / v%, and more preferably 0.001 to 0.03 w / v%.

  Furthermore, the ophthalmic composition of the present invention may contain a pharmacological component, if necessary. Examples of pharmacological ingredients that can be combined include, for example, sodium hyaluronate, diquafosol sodium, rebamipide, cyclosporine, and Liftelast, a dry eye therapeutic agent / keratoconjunctival epithelial disorder therapeutic agent; pranoprofen, dipotassium glycyrrhizinate, allantoin, epsilon. Aminocaproic acid, bromfenac sodium, ketorolac tromethamine, nepafenac, berberine chloride, berberine sulfate, sodium azulenesulfonate, zinc sulfate, zinc lactate, lysozyme hydrochloride and other anti-inflammatory agents; chlorpheniramine, chlorpheniramine salts (Chlorpheniramine maleate, etc.), diphenhydramine hydrochloride, and other antihistamines; antiallergic agents, such as sodium cromoglycate, ketotifen fumarate, acitazanolast, amlexanox, pemirolast potassium, tranilast, ibudilast, olopatadine hydrochloride; Antibacterial agents such as norfloxacin, ofloxacin, lomefloxacin, levofloxacin, gentamicin, gatifloxacin, azithromycin; ascorbic acid, flavin adenine dinucleotide sodium, cyanocobalamin, pyridoxine, pyridoxine salts (pyridoxine hydrochloride, etc.), tocopherol acetate, retinol acetate , Vitamins such as retinol palmitate, panthenol, calcium pantothenate, sodium pantothenate; aspartic acid, salts of aspartic acid (sodium aspartate, potassium aspartate, magnesium aspartate, etc.), taurine, sodium chondroitin sulfate, etc. Amino acids; monosaccharides such as glucose (glucose), galactose, mannose, fructose; anticholinesterase agents such as neostigmine and neostigmine salts (neostigmine methyl sulfate); naphazoline, tetrahydrozoline, epinephrine, ephedrine, phenylephrine, dl-methylephedrine. Vasoconstrictors such as sulfadiazine, sulfisoxazole, sulfisomidine, sulfadimethoxine, sulfamethoxypyridazine, sulfamethoxazole, sulfaetidol, sulfamethomidine, sulfaphenazole, sulfaguanidine, phthalylsulfate Sulfa drugs such as fathiazole and succinylsulfathiazole; latanoprost, brinzolamide, timolol maleate, bimatoprost, brimonidine tartrate And the like; antiviral agents such as idoxuridine and the like. The compounds exemplified herein may be in the form of salts or other salts, provided that they are pharmaceutically acceptable. These pharmacological components may be used alone or in combination of two or more.

  Concentrations of these pharmacological components may be appropriately set within a range satisfying the above-mentioned osmotic pressure, depending on the type of the pharmacological component to be used, the type and concentration of other components to be blended, and the like.

  Among these pharmacological components, preferred examples include at least one selected from the group consisting of pyridoxine, chlorpheniramine, cyanocobalamin, aspartic acid, taurine, neostigmine, and salts thereof.

  When pyridoxine and / or a salt thereof is contained in the ophthalmic composition of the present invention, the concentration thereof is not particularly limited, but is, for example, 0.001 to 0.3 w / v%, preferably 0.001 to 0. 2 w / v%, and more preferably 0.01 to 0.1 w / v%.

  When the ophthalmic composition of the present invention contains chlorpheniramine and / or a salt thereof, the concentration thereof is not particularly limited, but is, for example, 0.0006 to 0.1 w / v%, preferably 0.0006. -0.05 w / v%, More preferably, it is 0.006-0.03 w / v%.

  When cyanopobalamin is contained in the ophthalmic composition of the present invention, its concentration is not particularly limited, but for example, 0.0002 to 0.06 w / v%, preferably 0.0002 to 0.04 w / v%. , And more preferably 0.004 to 0.02 w / v%.

  When the ophthalmic composition of the present invention contains aspartic acid and / or a salt thereof, the concentration thereof is not particularly limited, but is, for example, 0.01 to 3 w / v%, preferably 0.01 to 2 w / v%, and more preferably 0.1 to 1 w / v%.

  When taurine is contained in the ophthalmic composition of the present invention, the concentration thereof is not particularly limited, but is, for example, 0.01 to 3 w / v%, preferably 0.01 to 2 w / v%, more preferably , 0.1 to 1 w / v%.

  When neostigmine and / or its salt is contained in the ophthalmic composition of the present invention, the concentration thereof is not particularly limited, but is, for example, 0.001 to 0.02 w / v%, preferably 0.001 to 0. 0.01 w / v%, and more preferably 0.001 to 0.005 w / v%.

  When glucose is contained in the ophthalmic composition of the present invention, the concentration thereof is not particularly limited, but is, for example, 0.0001 to 0.5 w / v%, preferably 0.0005 to 0.2 w / v%. , And more preferably 0.001 to 0.1 w / v%.

  By satisfying a predetermined osmotic pressure, the ophthalmic composition of the present invention does not need to contain a therapeutic agent for keratoconjunctival epithelial disorders such as hyaluronic acid and its alkali metal salts (sodium salt, potassium salt, etc.) as a pharmacological component. , Can promote healing of corneal epithelial wounds. Therefore, one embodiment of the ophthalmic composition of the present invention is that it does not contain hyaluronic acid or its alkali metal salt.

[PH]
The pH of the ophthalmic composition of the present invention is not particularly limited as long as it can be applied to the ocular mucosa, but examples thereof include pH 3 to 10, preferably pH 4 to 9, and more preferably pH 5 to 8.

[Formulation form]
The formulation of the ophthalmic composition of the present invention may be any as long as it contains water as a base, and may be, for example, an aqueous solution, a suspension, an emulsion, or the like, but preferably an aqueous solution. Can be mentioned.

  Further, the ophthalmic composition of the present invention may be in the form of an eye drop or an eye wash, but is preferably an eye drop.

[Usage / Dose / Usage]
The ophthalmic composition of the present invention is used for promoting the healing of corneal epithelial wounds. In the ophthalmic composition of the present invention, the corneal epithelial wound targeted for healing promotion may be caused by either an internal factor or an external factor. In the ophthalmic composition of the present invention, specific examples of the corneal epithelial wound targeted for healing promotion are as exemplified in the section “1. Definition” above. Among the corneal epithelial wounds targeted for healing promotion, preferably, the corneal epithelial defect associated with an endogenous disease such as allergic conjunctivitis or dry eye; the corneal epithelial defect associated with an extrinsic disease such as drug-induced or contact lens wear Can be mentioned.

  The ophthalmic composition of the present invention is used by applying to an eye having a corneal epithelial wound. For example, when the ophthalmic composition of the present invention is an eye drop, it may be applied to the eye having a corneal epithelial wound at 1 to 3 drops per day and 1 to 6 times per day. When the ophthalmic composition of the present invention is an eyewash solution, it may be washed 1 to 6 times a day by using 1 to 30 mL per time for an eye with a corneal epithelial wound.

[Production method]
The ophthalmic composition of the present invention may be produced according to a method known per se, and can be produced, for example, by the method described in the "General Rules for Formulations" section of the 17th Revised Japanese Pharmacopoeia.

4. Method for providing healing promoting action on corneal epithelial wound As described above, the osmotic pressure of the ophthalmic composition can be adjusted to 200 to 260 mOsm to provide the ophthalmic composition with a promoting action on healing of corneal epithelial wound. Therefore, as one aspect, a method for imparting a corneal epithelial wound healing promoting action to an ophthalmic composition, which comprises a step of adjusting the osmotic pressure of the ophthalmic composition to 200 to 260 mOsm, is provided.

  In the present method, the preferable range of osmotic pressure, the kind and concentration of components to be incorporated in the ophthalmic composition, the formulation form of the ophthalmic composition, the use, the dose, the usage, etc. are described in the above "3. Ophthalmic composition". Is as described in the column.

  Hereinafter, the present invention will be specifically described with reference to examples, but the present invention is not limited thereto.

Test Example 1: Investigation of corneal epithelial wound healing promoting action by scratch assay (1)
1. Test material and test method
1-1. cell
Human corneal epithelial cells immortalized with the V40-adenovirus recombinant vector (RIKEN cell bank, resource name RCB2280, Japan) were used.

1-2. Test medium As a growth medium, human insulin (human, recombinant, Fujifilm Wako Pure Chemical Industries, Ltd.) 5 μg / mL, penicillin / streptomycin 1 w / v%, fetal bovine serum (Heat inactivated FBS) (Thermo Fisher Scientific) 5 w / A DMEM / F12 medium containing v% and 10 ng / mL epidermal growth factor (EGF) (Pepro Tech) was prepared. Separately, sodium chloride as an osmotic pressure adjusting solution is 0.359 w / v%, 0.422 w / v%, 0.485 w / v%, 0.548 w / v%, 0.642 w / v%, and 0. An aqueous solution containing 0.837 w / v% was prepared.

  The growth medium and the osmotic pressure adjusting solution were mixed at a volume ratio of 1: 1 to prepare a test medium whose osmotic pressure was adjusted to 210 mOsm, 219 mOsm, 228 mOsm, 240 mOsm, 255 mOsm and 285 mOsm.

1-3. Test Method Cultured immortalized human corneal epithelial cells were diluted to 1.5 × 10 ⁵ cells / mL with a growth medium, and seeded in 24-well plates (Corning) at 1 mL per well. Immortalized human corneal epithelial cells were attached to the bottom of the well in a subconfluent state by incubating at 37 ° C. and 5% CO ₂ for about 24 hours. After the incubation, the medium in the well was removed with an aspirator, and 1 mL of phosphate buffer (DPBS) (Thermo Fisher Scientific) was added to the well. Then, the phosphate buffer in the well was removed with an aspirator, and scratches were performed using Cell Scratcher (ACG Technoglass) to prepare a cell defect region (width about 1.8 mm, length about 13 mm). After preparation of the cell-deficient region, 1 mL of phosphate buffer (DPBS) (Thermo Fisher Scientific) was added. After that, the phosphate buffer was removed with an aspirator, and 1 mL of each osmotic test medium was added. The time when the preparation of the cell-deficient region was completed was defined as the test start time (0 hour), and the extent of repair of the cell-deficient region was evaluated by quantifying the width of the cell-defective region 5 hours later. Specifically, using the all-in-one fluorescence microscope BZ-X700 (KEYENCE), the cell deficient area was photographed at 0 hours and 5 hours, and the photographed pictures were saved as a digital image in a computer, and image analysis software (Image- The width (pixel) of the cell deficient region was measured using Pro ^R Plus ver. 7.0J). The wound healing width (pixel) was calculated by subtracting the width (pixel) of the cell-deficient region after 5 hours from the width (pixel) of the cell-deficient region at 0 hour. In addition, the ratio of the wound healing width under each osmotic pressure condition to the wound healing width under isotonic condition (285 mOsm) (% _{vs isotonic condition} ) was also calculated. In this test, in one measurement (one 24-well plate), measurement in the same test medium was carried out in 4 wells. In addition, this test was performed 3 times in total in different well plates (n = 11 to 12).

2. Test Results The wound healing width under each osmotic pressure condition is shown in Table 1, and the approximate curve of the osmotic pressure and the wound healing width is shown in FIG. As a result, it was confirmed that in the low osmotic pressure range of 200 to 260 mOsm, the wound healing width showed a larger value than in the case of 285 mOsm which is isotonic. That is, the present results revealed that the wound healing of the corneal epithelium is promoted under the low osmotic pressure condition of 200 to 260 mOsm.

Test Example 2: Examination of corneal epithelial wound healing promoting action by scratch assay (2)
As the osmotic pressure adjusting liquid, an aqueous solution containing 1.173 w / v%, 1.786 w / v%, and 2.329 w / v% of glycerin was prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting solution were mixed at a volume ratio of 1: 1 to prepare a test medium having an osmotic pressure adjusted to 220 mOsm, 258 mOsm, and 288 mOsm.

Using the test medium, a test was conducted under the same conditions as in Test Example 1 (n = 6 to 7) to determine the wound healing width, and the osmotic pressure conditions for the wound healing width under the isotonic condition (288 mOsm) were measured. The ratio of wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 2 shows the wound healing width under each osmotic pressure condition. As a result, as in the case of Test Example 1, in the low osmotic pressure range of 200 to 260 mOsm, as compared with the case of 288 mOsm that is isotonic, the wound healing width shows a large value, and 200 to 260 mOsm. It was confirmed that corneal epithelial wound healing was promoted under hypotonic conditions.

  That is, it was revealed that the use of glycerin as the osmotic pressure adjusting agent promotes the wound healing of the corneal epithelium under the low osmotic pressure condition of 200 to 260 mOsm as in the case of using sodium chloride. ..

Test Example 3: Examination of corneal epithelial wound healing promoting action by scratch assay (3)
As an osmotic pressure adjusting liquid, an aqueous solution containing 0.989 w / v%, 1.506 w / v%, and 1.963 w / v% propylene glycol was prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting solution were mixed at a volume ratio of 1: 1 to prepare a test medium having an osmotic pressure adjusted to 220 mOsm, 255 mOsm, and 286 mOsm.

Using the test medium, a test was performed under the same conditions as in Test Example 1 (n = 6 to 8) to determine the wound healing width, and the osmotic pressure conditions for the wound healing width under the isotonic condition (286 mOsm) were measured. The ratio of wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 3 shows the width of wound healing under each osmotic pressure condition. As a result, as in the case of Test Examples 1 and 2, in the low osmotic pressure range of 200 to 260 mOsm, the wound healing width shows a large value as compared to the case of 286 mOsm which is isotonic, and 200 to 260 mOsm. It was confirmed that the wound healing of the corneal epithelium was promoted under the low osmotic pressure condition of 260 mOsm.

  That is, it is clear that even when propylene glycol is used as an osmotic pressure adjusting agent, corneal epithelial wound healing is promoted under a low osmotic pressure condition of 200 to 260 mOsm as in the case of using sodium chloride or glycerin. Became.

Test Example 4: Investigation of corneal epithelial wound healing promoting action by scratch assay (4)
As the osmotic pressure adjusting liquid, aqueous solutions A to F having the compositions shown in Table 4 were prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting liquids A to F were mixed at a volume ratio of 1: 1, and the test medium whose osmotic pressure was adjusted to 219 mOsm, 255 mOsm, 285 mOsm, 220 mOsm, 254 mOsm, and 285 mOsm was used. Prepared.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 14 to 16) to determine the wound healing width, and the osmotic pressure conditions for the wound healing width under the isotonic condition (285 mOsm) are measured. The ratio of wound healing width (% _{vs. isotonic condition} ) was calculated. The ratio under the conditions using the aqueous solutions A and B was calculated in comparison with the isotonic condition adjusted using the aqueous solution C, and the ratio under the conditions using the aqueous solutions D and E was the aqueous solution F. It was calculated in comparison with the isotonic condition adjusted by.

  The wound healing width under each osmotic pressure condition is shown in Table 5. As a result, even if the osmotic pressure is adjusted using glycerin, boric acid, and borax, it is isotonic in the low osmotic pressure range of 200 to 260 mOsm as in the case of Test Examples 1 to 3. The wound healing width was larger than that of 285 mOsm, and it was confirmed that the wound healing of the corneal epithelium was promoted under the low osmotic pressure condition of 200 to 260 mOsm.

Test Example 5: Examination of corneal epithelial wound healing promoting action by scratch assay (5)
As the osmotic pressure adjusting liquid, aqueous solutions G to I having the compositions shown in Table 6 were prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting liquids G to I were mixed at a volume ratio of 1: 1 to prepare a test medium having an osmotic pressure adjusted to 222 mOsm, 254 mOsm, and 283 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 16) to determine the wound healing width, and the wound healing under each osmotic pressure condition with respect to the wound healing width under the isotonic condition (283 mOsm). The width ratio (% _{vs. isotonic condition} ) was calculated.

  The wound healing width under each osmotic pressure condition is shown in Table 7. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting it in the low osmotic pressure range of 200 to 260 mOsm even if it contained sodium edetate hydrate.

Test Example 6: Examination of corneal epithelial wound healing promoting action by scratch assay (6)
As the osmotic pressure adjusting liquid, aqueous solutions J and K having the compositions shown in Table 8 were prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting solutions J and K were mixed at a volume ratio of 1: 1 to prepare test mediums having osmotic pressures adjusted to 218 mOsm and 284 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (218 mOsm) for the wound healing width under the isotonic condition (284 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 9 shows the width of wound healing under each osmotic pressure condition. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting it in the range of low osmotic pressure of 200 to 260 mOsm even when 1-menthol was included.

Test Example 7: Examination of corneal epithelial wound healing promoting action by scratch assay (7)
As the osmotic pressure adjusting liquid, aqueous solutions L and M having the compositions shown in Table 10 were prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting solutions L and M were mixed at a volume ratio of 1: 1 to prepare test mediums having osmotic pressures adjusted to 221 mOsm and 286 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (221 mOsm) for the wound healing width under isotonic conditions (286 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 11 shows the width of wound healing under each osmotic pressure condition. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting the range of the hypoosmotic pressure of 200 to 260 mOsm even if the pyridoxine hydrochloride was contained.

Test Example 8: Examination of corneal epithelial wound healing promoting action by scratch assay (8)
Aqueous solutions N and O having the compositions shown in Table 12 were prepared as osmotic pressure adjusting liquids. The growth medium used in Test Example 1 and the osmotic pressure adjusting solutions N and O were mixed at a volume ratio of 1: 1 to prepare test mediums having osmotic pressures adjusted to 224 mOsm and 287 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (224 mOsm) for the wound healing width under an isotonic condition (287 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 13 shows the wound healing width under each osmotic pressure condition. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting it in the low osmotic pressure range of 200 to 260 mOsm even if it contained trometamol.

Test Example 9: Investigation of corneal epithelial wound healing promoting action by scratch assay (9)
As the osmotic pressure adjusting liquid, aqueous solutions P and Q having the compositions shown in Table 14 were prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting liquids P and Q were mixed at a volume ratio of 1: 1 to prepare a test medium whose osmotic pressure was adjusted to 221 mOsm and 287 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (221 mOsm) for the wound healing width under isotonic condition (287 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 15 shows the wound healing width under each osmotic pressure condition. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting it in the low osmotic pressure range of 200 to 260 mOsm even when the sodium citrate hydrate was included.

Test Example 10: Examination of corneal epithelial wound healing promoting action by scratch assay (10)
Aqueous solutions R and S having the compositions shown in Table 16 were prepared as osmotic pressure adjusting liquids. The growth medium used in Test Example 1 and the osmotic pressure adjusting solutions R and S were mixed at a volume ratio of 1: 1 to prepare test mediums having osmotic pressures adjusted to 224 mOsm and 286 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (224 mOsm) for the wound healing width under isotonic condition (286 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 17 shows the wound healing width under each osmotic pressure condition. As a result, it was confirmed that the wound healing of corneal epithelium was promoted by setting the hypotonic range of 200 to 260 mOsm even when the disodium hydrogen phosphate dodecahydrate was included.

Test Example 11: Investigation of corneal epithelial wound healing promoting action by scratch assay (11)
Aqueous solutions T and U having the compositions shown in Table 18 were prepared as osmotic pressure adjusting liquids. The growth medium used in Test Example 1 and the osmotic pressure adjusting liquids T and U were mixed at a volume ratio of 1: 1 to prepare a test medium whose osmotic pressure was adjusted to 225 mOsm and 286 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (225 mOsm) for the wound healing width under isotonic condition (286 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 19 shows the wound healing width under each osmotic pressure condition. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting the hypotonic range of 200 to 260 mOsm even if glucose was included.

Test Example 12: Investigation of corneal epithelial wound healing promoting action by scratch assay (12)
Aqueous solutions V and W having the compositions shown in Table 20 were prepared as osmotic pressure adjusting liquids. The growth medium used in Test Example 1 and the osmotic pressure adjusting solutions V and W were mixed at a volume ratio of 1: 1 to prepare test mediums having osmotic pressures adjusted to 224 mOsm and 287 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (224 mOsm) for the wound healing width under an isotonic condition (287 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 21 shows the wound healing width under each osmotic pressure condition. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting it in the range of low osmotic pressure of 200 to 260 mOsm even if it contained chlorpheniramine maleate.

Test Example 13: Examination of corneal epithelial wound healing promoting action by scratch assay (13)
As the osmotic pressure adjusting liquid, aqueous solutions X and Y having the compositions shown in Table 22 were prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting liquids X and Y were mixed at a volume ratio of 1: 1 to prepare a test medium whose osmotic pressure was adjusted to 225 mOsm and 289 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (225 mOsm) for the wound healing width under an isotonic condition (289 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 23 shows the width of wound healing under each osmotic pressure condition. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting it in the low osmotic pressure range of 200 to 260 mOsm, even if it contained neostigmine methylsulfate.

Test Example 14: Examination of corneal epithelial wound healing promoting action by scratch assay (14)
As the osmotic pressure adjusting liquid, aqueous solutions AA and BB having the compositions shown in Table 24 were prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting solutions AA and BB were mixed at a volume ratio of 1: 1 to prepare test mediums having osmotic pressures adjusted to 219 mOsm and 286 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (219 mOsm) for the wound healing width under isotonic conditions (286 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 25 shows the wound healing width under each osmotic pressure condition. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting the low osmotic pressure range of 200 to 260 mOsm even if taurine was contained.

Test Example 15: Examination of corneal epithelial wound healing promoting action by scratch assay (15)
As the osmotic pressure adjusting liquid, aqueous solutions CC and DD having the compositions shown in Table 26 were prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting liquids CC and DD were mixed at a volume ratio of 1: 1 to prepare a test medium whose osmotic pressure was adjusted to 224 mOsm and 287 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (224 mOsm) for the wound healing width under an isotonic condition (287 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 27 shows the wound healing width under each osmotic pressure condition. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting it in the range of low osmotic pressure of 200 to 260 mOsm even if it contained potassium L-aspartate.

Test Example 16: Examination of corneal epithelial wound healing promoting action by scratch assay (16)
As the osmotic pressure adjusting liquid, aqueous solutions EE and FF having the compositions shown in Table 28 were prepared. The growth medium used in Test Example 1 and the osmotic pressure adjusting liquids EE and FF were mixed at a volume ratio of 1: 1 to prepare a test medium whose osmotic pressure was adjusted to 224 mOsm and 290 mOsm.

Using the test medium, a test is performed under the same conditions as in Test Example 1 (n = 12) to determine a wound healing width, and a low osmotic pressure condition (224 mOsm) for the wound healing width under isotonic condition (290 mOsm). The ratio of the wound healing width (% _{vs. isotonic condition} ) was calculated.

  Table 29 shows the width of wound healing under each osmotic pressure condition. As a result, it was confirmed that the wound healing of the corneal epithelium was promoted by setting the range of low osmotic pressure of 200 to 260 mOsm even when cyanocobalamin was contained.

Comprehensive Consideration From the results of the above-mentioned Test Examples 1 to 16, corneal epithelium under low osmotic pressure conditions of 200 to 260 mOsm, irrespective of the types of components used for adjusting the osmotic pressure, the types of additives and pharmacological components to be blended, etc. It was revealed that the wound healing of the mouse was promoted.

  In Test Examples 1 to 16, due to different test dates, due to the difference in the state of the immortalized human corneal epithelial cells used in each test, the wound healing was performed at the same osmotic pressure. The width may show different values. However, in all the test examples, it is shown that the wound healing width increases under low osmotic pressure (200 to 260 mOsm) conditions as compared with the case of isotonicity, and the wound of corneal epithelium is observed under the low osmotic pressure condition. It can be said that healing is accelerated.

Test Example 17: Examination of corneal epithelial wound healing promoting action in vivo By the following tests, it can be confirmed that the ophthalmic composition of the present invention exhibits a corneal epithelial wound healing promoting action in vivo.

1. Test material and test method
1-1. Animals used Male white Japanese rabbits are used. Regarding the use of experimental animals, the Animal Experiment Ethics Committee based on the Act on the Protection and Management of Animals (Law No. 105, October 1, 1973, and the final revision, Law No. 38, June 12, 2013) Approve and implement.

1-2. Test Eye Drops Isotonic eye drops and hypotonic eye drops having the compositions shown in Table 30 are prepared as test eye drops.

1-3. Test method 1) Corneal epithelial curettage 50 mg / mL ketamine injection solution (Ketaral ^R intramuscular injection 500 mg; Daiichi Sankyo Propharma) and 20 mg / mL seralactal injection solution (Ceralactal ^R 2% injection solution; Bayer Yakuhin) 3: 1 After anesthetizing the animal by intramuscular injection (1 mL / kg) of the mixed solution, 0.4 w / v% oxybuprocaine hydrochloride ophthalmic solution (Benoxir ^R ophthalmic solution 0.4%; Santen Pharmaceutical) Local anesthesia is administered using. Then, after dislocating the eyeball, a corneal epithelium in the central portion of the cornea was engraved with a diameter of 10 mm using a trepan with a diameter of 10 mm, and the corneal epithelium in the circumference was engraved with a handy router under a stereoscopic microscope. Cure the layers. After the curettage, the corneal surface is washed with a physiological saline solution (Otsuka raw food injection; Otsuka Pharmaceutical Factory), and the eyeball is repositioned in the orbit to complete the corneal epithelial curettage treatment.

2) Administration of test ophthalmic solution Test ophthalmic solution was administered twice a day on the day of corneal epithelial curettage, 4 times a day after curettage 1 and 2 days, and once a day 3 days after curettage, each at intervals of 2 hours or more. And the eye is treated with a micropipette at a dose of 50 μL once for the treated eye.

3) Area measurement of corneal epithelial defect part The time when the corneal epithelial scraping is completed is defined as the test start time (0 hour), and 48 hours after that, the extent of corneal epithelial repair is evaluated by quantifying the corneal epithelial defect area. Specifically, after 48 hours, 10 μL of 0.1 w / v% fluorescein sodium (Fuji Film Wako Pure Chemical Industries, Ltd.) solution was instilled into the treated eye, and immediately a slit lamp was used to take a photograph of the anterior segment of the animal. The fluorescein-stained corneal epithelial defect area is recorded by photographing. The developed photograph was saved as a digital image in a computer, and the area of the corneal epithelial defect stained with fluorescein was measured using image analysis software (Image-Pro ^R Plus ver. 7.0J), and the residual wound after 48 hours was measured. Calculate the area ratio.

Claims (7)

  An ophthalmic composition for promoting corneal epithelial wound healing, which has an osmotic pressure of 200 to 260 mOsm.   The ophthalmic composition for promoting corneal epithelial wound healing according to claim 1, which has an osmotic pressure of 220 to 260 mOsm.   The ophthalmic composition for promoting corneal epithelial wound healing according to claim 1 or 2, which comprises an alkali metal salt and / or a polyhydric alcohol.   The ophthalmic composition for promoting corneal epithelial wound healing according to claim 3, wherein the alkali metal salt is sodium chloride.   The ophthalmic composition for promoting corneal epithelial wound healing according to claim 3, wherein the polyhydric alcohol is propylene glycol and / or glycerin.   The ophthalmic composition for promoting corneal epithelial wound healing according to claim 1, which is an eye drop.   A method for imparting a corneal epithelial wound healing promoting action to an ophthalmic composition, which comprises the step of adjusting the osmotic pressure of the ophthalmic composition to 200 to 260 mOsm.