JP2023067803A - Ophthalmologic composition - Google Patents

Abstract

To provide a new ophthalmologic composition that can protect cornea.SOLUTION: An ophthalmologic composition for corneal protection comprises at least one selected from the group consisting of chondroitin sulfate with a weight average molecular weight of 40,000-70,000 and salts thereof.SELECTED DRAWING: None

Description

The present invention relates to ophthalmic compositions.

Chondroitin sulfate or its salt is a type of acidic mucopolysaccharide, and is used in ophthalmic preparations for the purpose of promoting energy metabolism, promoting metabolism and cell respiration to relieve eye fatigue, and replenishing tear components. are blended (for example, Patent Literature 1).

JP 2011-148791 A

An object of the present invention is to provide a new ophthalmic composition capable of protecting the cornea.

The present inventors unexpectedly found that sodium chondroitin sulfate having a specific weight-average molecular weight has a protective effect on the cornea.

The present invention provides, for example, the following inventions.
[1]
An ophthalmic composition for protecting the cornea, containing at least one selected from the group consisting of chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and salts thereof.
[2]
An ophthalmic composition for repairing or recovering corneal damage, containing at least one selected from the group consisting of chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and salts thereof.
[3]
A corneal protective agent containing at least one selected from the group consisting of chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and salts thereof.
[4]
A corneal injury repairing agent or injury recovery agent containing at least one selected from the group consisting of chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and salts thereof.
[5]
A method for imparting a corneal-protective effect to an ophthalmic composition, comprising incorporating at least one selected from the group consisting of chondroitin sulfate and salts thereof having a weight-average molecular weight of 40,000 to 70,000 into the ophthalmic composition.
[6]
The ophthalmic composition contains at least one selected from the group consisting of chondroitin sulfate having a weight-average molecular weight of 40,000 to 70,000 and salts thereof. How to give effect.

ADVANTAGE OF THE INVENTION According to this invention, the ophthalmic composition which can protect a cornea can be provided. Further, according to the present invention, it is possible to provide an ophthalmic composition capable of repairing or recovering corneal damage.

4 is a graph showing the results of Test Example 1. FIG. 4 is a graph showing the results of Test Example 2. FIG. 4 is a graph showing the results of Test Example 3. FIG. 4 is a graph showing the results of gene expression of occludin in Test Example 4. FIG. 10 is a graph showing the results of gene expression of membrane-type mucin in Test Example 4. FIG. 10 is a photograph showing the results of immunostaining for occludin in Test Example 5. FIG. FIG. 10 is a photograph showing the results of immunostaining for membrane-type mucin in Test Example 5. FIG.

DETAILED DESCRIPTION OF THE INVENTION Embodiments for carrying out the present invention will be described in detail below. However, the present invention is not limited to the following embodiments.

In this specification, unless otherwise specified, the content unit "%" means "w/v %" and is synonymous with "g/100 mL".

The ophthalmic composition according to the present embodiment contains at least one selected from the group consisting of chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and salts thereof (also simply referred to as "component (A)"). contains.

Chondroitin sulfate and salts thereof are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutical) or physiologically acceptable.

Salts of chondroitin sulfate include, for example, alkali metal salts and alkaline earth metal salts. Examples of alkali metal salts include sodium salts and potassium salts. Examples of alkaline earth metal salts include magnesium salts and calcium salts.

Chondroitin sulfate and salts thereof are preferably chondroitin sulfate and alkali metal salts of chondroitin sulfate, more preferably chondroitin sulfate and sodium chondroitin sulfate, and still more preferably sodium chondroitin sulfate.

Chondroitin sulfate and its salts may be natural products or synthetic products, but are generally natural products of animals (preferably mammals, fish, mollusks, etc.; more preferably bovine, shark, squid, A chondroitin sulfate derived from a ray, etc.) and a salt thereof are preferably used, a chondroitin sulfate derived from a shark and/or a ray and a salt thereof are more preferably used, and a chondroitin sulfate and a salt thereof derived from a shark are more preferably used. .

Commercially available chondroitin sulfate and salts thereof can also be used. Chondroitin sulfate and its salt may be used alone or in combination of two or more. In addition, chondroitin sulfate and its salts include chondroitin sulfate and its salts having a weight-average molecular weight of 40,000 to 70,000 defined herein and chondroitin sulfate and its salts having a weight-average molecular weight outside the range defined herein. can be used in combination with For example, sodium chondroitin sulfate with a weight average molecular weight of 56,000 and sodium chondroitin sulfate with a weight average molecular weight of 25,000 can be used in combination. When using a combination of chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and a salt thereof defined herein and chondroitin sulfate having a weight average molecular weight outside the range defined herein and a salt thereof, , chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and a salt thereof may be contained as raw materials.

As used herein, the "weight average molecular weight" can be determined by using gel permeation chromatography in which a multi-angle light scattering detector (MALS detector) and a differential refractive index detector (RI detector) are connected online. . Specifically, the following conditions are presented.
<Standard sample preparation>
10 mL of 0.1 M sodium nitrate aqueous solution was added to 5 mg of chondroitin sulfate or its salt, and the mixture was gently stirred at room temperature to dissolve completely.
<Conditions for measuring weight average molecular weight>
Apparatus: Gel permeation chromatograph - multi-angle light scatterometer Detector: Differential refractive index detector (Optilab rEX manufactured by Wyatt Technology)
Multi-angle light scattering detector (DAWN HELEOS manufactured by Wyatt Technology)
Column: 2 Shodex OHpak SB-806M HQ (φ7.8 mm × 30 cm, manufactured by Showa Denko)
Solvent: 0.1 M sodium nitrate aqueous solution Flow rate: 0.7 mL/min
Column temperature: 23°C
Detector temperature: 23°C
Injection volume: 0.2 mL
Data processing: Wyatt Technology data processing system (ASTRA)
The weight average molecular weight of chondroitin sulfate and its salt calculated by the above method is not particularly limited as long as it is in the range of 40,000 to 70,000. Examples of the lower limit of the weight average molecular weight include 41000 or more, 42000 or more, 43000 or more, 44000 or more, 45000 or more, 46000 or more, 47000 or more, 48000 or more, 49000 or more, and 50000 or more. Examples of the upper limit of the weight average molecular weight include 69000 or less, 68000 or less, 67000 or less, 66000 or less, 65000 or less, 64000 or less, 63000 or less, 62000 or less, 61000 or less, and 60000 or less. The weight average molecular weight ranges are 41,000 to 69,000, 42,000 to 68,000, 43,000 to 67,000, 44,000 to 66,000, 45,000 to 65,000, 46,000 to 64,000, 47,000 to 63,000, 48,000 to 62,000, 49,000 to 61,000, and 500. 00 to 60000 are examples be done.

The content of component (A) in the ophthalmic composition according to the present embodiment is not particularly limited, and is appropriately set according to the type and content of other compounding components, the application and formulation form of the ophthalmic composition, and the like. The content of component (A) is, for example, usually 0.001 to 5 w/v%, based on the total amount of the ophthalmic composition, and 0.005 to 5 w/ v%, more preferably 0.008 to 4 w/v%, even more preferably 0.01 to 3 w/v%, and 0.05 to 2 w/v% Even more preferably, 0.1 to 1 w/v% is particularly preferable, and 0.3 to 1 w/v% is most preferable.

The ophthalmic composition according to this embodiment may further contain a surfactant. When the ophthalmic composition further contains a surfactant, the effects of the present invention are exhibited more remarkably. Surfactants are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutical) or physiologically acceptable, nonionic surfactants, amphoteric surfactants, anionic surfactants , cationic surfactants.

Examples of nonionic surfactants include POE (20) sorbitan monolaurate (polysorbate 20), POE (20) sorbitan monopalmitate (polysorbate 40), POE (20) sorbitan monostearate (polysorbate 60), POE sorbitan fatty acid esters such as POE (20) sorbitan stearate (polysorbate 65), POE (20) sorbitan monooleate (polysorbate 80); POE (40) hydrogenated castor oil (polyoxyethylene hydrogenated castor oil 40), POE ( 60) POE hydrogenated castor oil such as hydrogenated castor oil (polyoxyethylene hydrogenated castor oil 60); POE (3) hydrogenated castor oil (polyoxyethylene castor oil 3), POE (10) castor oil (polyoxyethylene castor oil 10 ), POE castor oil such as POE (35) castor oil (polyoxyethylene castor oil 35); POE alkyl ether such as POE (9) lauryl ether; polyethylene glycol monostearate such as polyoxyl stearate 40; POE (20) POE-POP alkyl ethers such as POP(4) cetyl ether; copolymers, and the like. In the compounds exemplified above, POE is polyoxyethylene, POP is polyoxypropylene, and numbers in parentheses indicate the number of added moles.

Amphoteric surfactants include, for example, alkyldiaminoethylglycine or salts thereof (eg, hydrochlorides, etc.).

Examples of anionic surfactants include alkylbenzenesulfonates, alkylsulfates, polyoxyethylene alkylsulfates, aliphatic α-sulfomethyl esters, α-olefinsulfonic acids and the like.

Examples of cationic surfactants include cetylpyridinium chloride, benzalkonium chloride, benzethonium chloride and the like.

Among these surfactants, nonionic surfactants are preferred, and POE sorbitan fatty acid esters, POE hydrogenated castor oil, POE castor oil, and POE/POP block copolymers are more preferred. Commercially available surfactants can also be used. Surfactants may be used alone or in combination of two or more.

The content of the surfactant in the ophthalmic composition according to this embodiment is not particularly limited, and is appropriately set according to the type of surfactant, the application of the ophthalmic composition, the form of formulation, and the like. As for the content of the surfactant, from the viewpoint of exhibiting the effects of the present invention more remarkably, for example, the total content of the surfactant is 0.001 to 3 w/v% based on the total amount of the ophthalmic composition. It may be 0.005-2 w/v%, 0.01-1 w/v%, or 0.05-1 w/v%.

In the ophthalmic composition according to the present embodiment, the content ratio of the surfactant to the component (A) is not particularly limited. It is appropriately set according to the use of the product, the formulation form, and the like. From the viewpoint of further enhancing the effect of the present invention, the content ratio of the surfactant to the component (A) is, for example, 1 part by mass of the total content of the component (A) contained in the ophthalmic composition according to the present embodiment. On the other hand, even if the total content of the surfactant is 0.001 to 30 parts by mass, 0.005 to 20 parts by mass, 0.01 to 10 parts by mass, or 0.01 to 1 part by mass good.

The ophthalmic composition according to this embodiment preferably further contains a buffering agent. When the ophthalmic composition further contains a buffering agent, the effects of the present invention are exhibited more remarkably. The buffering agent is not particularly limited as long as it is pharmaceutically, pharmacologically (pharmaceutical) or physiologically acceptable. Buffers include, for example, inorganic buffers that are buffers derived from inorganic acids, and organic buffers that are buffers derived from organic acids or organic bases.

Examples of inorganic buffers include borate buffers, phosphate buffers, carbonate buffers and the like. Boric acid buffers include boric acid or salts thereof (alkali metal borates, alkaline earth metal borates, etc.). Phosphate buffers include phosphoric acid and salts thereof (alkali metal phosphate, alkaline earth metal phosphate, etc.). Carbonic acid buffers include carbonic acid or salts thereof (alkali metal carbonates, alkaline earth metal carbonates, etc.). Borate, phosphate, or carbonate hydrates may also be used as the borate buffer, phosphate buffer, or carbonate buffer. More specific examples include boric acid or a salt thereof (sodium borate, potassium tetraborate, potassium metaborate, ammonium borate, borax, etc.) as a borate buffer; Salts (disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, trisodium phosphate, tripotassium phosphate, calcium monohydrogen phosphate, calcium dihydrogen phosphate, etc.); Alternatively, salts thereof (sodium hydrogen carbonate, sodium carbonate, ammonium carbonate, potassium carbonate, calcium carbonate, potassium hydrogen carbonate, magnesium carbonate, etc.) can be exemplified.

Examples of organic buffers include citrate buffers, acetate buffers, lactate buffers, succinate buffers, Tris buffers, AMPD buffers and the like. Citric acid buffers include citric acid or salts thereof (alkali metal citrate, alkaline earth metal citrate, etc.). The acetate buffer includes acetic acid or salts thereof (alkali metal acetate, alkaline earth metal acetate, etc.). Lactic acid buffers include lactic acid or salts thereof (alkali metal lactate, alkaline earth metal lactate, etc.). Succinic acid buffers include succinic acid or salts thereof (such as alkali metal succinate). In addition, hydrates of citrate, acetate, lactate, or succinate may be used as the citrate buffer, acetate buffer, lactate buffer, or succinate buffer. More specific examples include citric acid or a salt thereof (sodium citrate, potassium citrate, calcium citrate, sodium dihydrogen citrate, disodium citrate, etc.) as a citric acid buffer; acetic acid as an acetic acid buffer or salts thereof (ammonium acetate, sodium acetate, potassium acetate, calcium acetate, etc.); lactic acid or salts thereof (sodium lactate, potassium lactate, calcium lactate, etc.) as a lactic acid buffer; monosodium succinate, disodium succinate, etc.). Tris buffers include, for example, trometamol or salts thereof (trometamol hydrochloride, etc.). AMPD buffers include, for example, 2-amino-2-methyl-1,3-propanediol or salts thereof.

As the buffering agent, a boric acid buffer (for example, a combination of boric acid and borax), boric acid and its salts are more preferred, and a combination of boric acid and borax is even more preferred.

A commercially available buffer may be used. A buffering agent may be used individually by 1 type, or may be used in combination of 2 or more types.

The content of the buffering agent in the ophthalmic composition according to the present embodiment is not particularly limited, and is appropriately set according to the type of buffering agent, the type and content of other compounding ingredients, the application and formulation form of the ophthalmic composition, and the like. be. The content of the buffering agent is, for example, 0.01 to 10 w/v%, 0.01 to 10 w/v%, based on the total amount of the ophthalmic composition, from the viewpoint of exhibiting the effects of the present invention more remarkably. 05-5 w/v%, or 0.1-3 w/v%.

In the ophthalmic composition according to the present embodiment, the content ratio of the buffering agent to the component (A) is not particularly limited. It is appropriately set according to the application, dosage form, and the like. From the viewpoint of further enhancing the effects of the present invention, the content ratio of the buffering agent to component (A) is, for example, In addition, the total content of buffering agents may be 0.01 to 100 parts by weight, 0.05 to 50 parts by weight, or 0.1 to 30 parts by weight.

The ophthalmic composition according to this embodiment may further contain neostigmine and salts thereof. Neostigmine and its salts are not particularly limited as long as they are pharmaceutically, pharmacologically (pharmaceutical) or physiologically acceptable. Salts of neostigmine include, for example, neostigmine methyl sulfate. As neostigmine and its salts, neostigmine methyl sulfate is preferred.

Commercially available neostigmine and salts thereof can also be used. Neostigmine and salts thereof may be used singly or in combination of two or more.

The content of neostigmine and salts thereof in the ophthalmic composition according to the present embodiment is 0.0001 to 0.05 w/v% based on the total amount of the ophthalmic composition. is preferably 0.0005 to 0.01 w/v%, more preferably 0.0008 to 0.008 w/v%, and 0.001 to 0.005 w/v% is more preferred.

The content ratio of neostigmine and its salt to the component (A) in the ophthalmic composition according to the present embodiment is not particularly limited. It is appropriately set according to the use, formulation form, etc. of the ophthalmic composition. The content ratio of neostigmine and its salt to the component (A) is, from the viewpoint of further enhancing the effect of the first present invention, for example, the total content of the component (A) contained in the ophthalmic composition according to the present embodiment is 1 The total content of neostigmine and its salts is preferably 0.0001 to 5 parts by mass, more preferably 0.0001 to 1 part by mass, and 0.0005 to 0.8 parts by mass. It is more preferably 0.001 to 0.5 parts by mass, particularly preferably 0.001 to 0.05 parts by mass, and 0.005 to 0.025 parts by mass. is most preferred.

The ophthalmic composition according to this embodiment may further contain vitamin A. Vitamin A is not particularly limited as long as it is pharmaceutically, pharmacologically (pharmaceutical) or physiologically acceptable. Specific examples of vitamin A include retinol, retinal, retinoic acid, derivatives thereof, and salts thereof.

Examples of vitamin A derivatives include esters with monovalent carboxylic acids such as retinol palmitate, retinol acetate, retinol butyrate, retinol propionate, retinol octylate, retinol laurate, retinol oleate and retinol linolenate. mentioned.

Examples of vitamin A salts include organic acid salts [e.g., monocarboxylates (acetate, trifluoroacetate, butyrate, palmitate, stearate, etc.), polyvalent carboxylates (fumaric acid salt, maleate, succinate, malonate, etc.), oxycarboxylate (lactate, tartrate, citrate, etc.), organic sulfonate (methanesulfonate, toluenesulfonate, tosylate salts, etc.)], inorganic acid salts (e.g., hydrochlorides, sulfates, nitrates, hydrobromides, phosphates, etc.), salts with organic bases (e.g., methylamine, triethylamine, triethanolamine, morpholine , piperazine, pyrrolidine, tripyridine, salts with organic amines such as picoline, etc.), salts with inorganic bases [e.g., ammonium salts; alkali metals (sodium, potassium, etc.), alkaline earth metals (calcium, magnesium, etc.), aluminum salts with metals such as].

The vitamin A is preferably a derivative of retinol, more preferably an ester of retinol and a monovalent carboxylic acid, more preferably retinol palmitate and retinol acetate, and even more preferably retinol palmitate.

As the vitamin A, synthetic products may be used, or extracts obtained from natural products (for example, vitamin A oil, etc.) may be used. Vitamin A oil is a fatty oil obtained from animal tissue containing retinol, a concentrate thereof, or a mixture thereof with vegetable oil added as appropriate. Commercially available vitamin A can also be used. Vitamin A may be used individually by 1 type, or may be used in combination of 2 or more type.

The content of vitamin A in the ophthalmic composition according to the present embodiment, based on the total amount of the ophthalmic composition, is 0.1 to 100,000 IU/100 mL, 0.5 to 75,000 IU/100 mL, Or it may be 10,000 to 50,000 IU/100 mL.

"IU" means an international unit determined by the method described in the Japanese Pharmacopoeia 17th Edition, Vitamin A Determination Method, etc. For example, in each article of the Japanese Pharmacopoeia 17th Edition, it is stated that retinol acetate contains 2.5 million units or more of vitamin A per 1g, and retinol palmitate contains 1.5 million units or more of vitamin A per 1g. ing.

In the ophthalmic composition according to the present embodiment, the content ratio of vitamin A relative to component (A) is not particularly limited. It is appropriately set according to the use of the product, the formulation form, and the like. From the viewpoint of further enhancing the effect of the third aspect of the present invention, the content ratio of the vitamin A relative to the component (A) is, for example, 1 mass of the total content of the component (A) contained in the ophthalmic composition according to the present embodiment. part, the total content of vitamin A may be 0.1 to 1,000,000 IU/g, 0.05 to 750,000 IU/g, or 10,000 to 500,000 IU/g .

The ophthalmic composition according to this embodiment may further contain menthol. Menthol is not particularly limited as long as it is pharmaceutically, pharmacologically (pharmaceutical) or physiologically acceptable.

Menthol may be d-, l- or dl-menthol, exemplified by l-menthol, d-menthol and dl-menthol. Essential oils containing menthol may also be used. Examples of such essential oils include mint oil, cool mint oil, spearmint oil, peppermint oil and the like.

The content of menthol in the ophthalmic composition according to the present embodiment is not particularly limited, and is appropriately set according to the type and content of other compounding ingredients, the application and formulation form of the ophthalmic composition, and the like. The content of menthol is 0.00001 to 0.5 w/v%, 0.0001 to 0.1 w/v%, or 0.001 to 0.05 w/v% based on the total amount of the ophthalmic composition. may When using an essential oil containing menthol, the blending ratio of the essential oil is set so that the content of menthol in the essential oil to be blended satisfies the above blending ratio.

In the ophthalmic composition according to the present embodiment, the content ratio of menthol to component (A) is not particularly limited, and the type of component (A), the type and content of other compounding components, the use and formulation form of the ophthalmic composition. etc., and is set as appropriate. The content ratio of menthol to the component (A) is, for example, 0.00001 to 0.00001 to 1 part by mass of the total content of the component (A) contained in the ophthalmic composition according to the present embodiment. It may be 10 parts by weight, 0.0001 to 1 part by weight, or 0.001 to 0.1 parts by weight.

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

The ophthalmic composition according to the present embodiment can be adjusted to have an osmotic pressure ratio within a range acceptable to living organisms, if necessary. An appropriate osmotic pressure ratio can be appropriately set according to the application, formulation form, method of use, etc. of the ophthalmic composition, and can be, for example, 0.4 to 5.0, and 0.6 to 3.0. , more preferably 0.8 to 2.2, even more preferably 0.8 to 2.0. The osmotic pressure ratio is the ratio of the osmotic pressure of the sample to 286 mOsm (osmotic pressure of 0.9 w/v% sodium chloride aqueous solution) based on the 17th revision of the Japanese Pharmacopoeia, and the osmotic pressure is the osmotic pressure measurement method described in the Japanese Pharmacopoeia. (freezing point depression method). The standard solution for osmotic pressure ratio measurement (0.9 w/v% sodium chloride aqueous solution) was obtained by drying sodium chloride (Japanese Pharmacopoeia standard reagent) at 500 to 650 ° C. for 40 to 50 minutes and then placing it in a desiccator (silica gel). After allowing to cool, 0.900 g thereof is accurately weighed and dissolved in purified water to prepare exactly 100 mL, or a commercially available standard solution for osmotic pressure ratio measurement (0.9 w/v % sodium chloride aqueous solution) can be used.

The viscosity of the ophthalmic composition according to this embodiment is not particularly limited as long as it is within a pharmaceutically, pharmacologically (pharmaceutical) or physiologically acceptable range. As the viscosity of the ophthalmic composition according to the present embodiment, for example, the viscosity at 20 ° C. measured with a rotational viscometer (TV-20 type viscometer, manufactured by Toki Sangyo Co., Ltd., rotor; 1 ° 34' × R24) is 1 It is preferably from 1 to 10,000 mPa s, more preferably from 1 to 8,000 mPa s, still more preferably from 1 to 1,000 mPa s, even more preferably from 1 to 100 mPa s, and from 1 to 20 mPa. · s is particularly preferred, and 1.5 to 10 mPa·s is most preferred.

The ophthalmic composition according to the present embodiment contains, in addition to the above components, an appropriate amount of a combination of components selected from various pharmacologically active components and physiologically active components within a range that does not impair the effects of the present invention. good too. The ingredients are not particularly limited, and examples thereof include active ingredients in ophthalmic drugs described in the 2017 version of the Approval Standards for Manufacturing and Marketing of OTC Drugs (supervised by the Japanese Society of Regulatory Science). Specific examples of components used in ophthalmic drugs include the following components.
Antiallergic agents: for example, cromoglycate sodium, tranilast, pemirolast potassium, acitazanolast, amlexanox, ibudilast and the like.
Antihistamines: e.g. diphenhydramine or salts thereof (e.g. diphenhydramine hydrochloride), iproheptine or salts thereof (e.g. iproheptine hydrochloride), chlorpheniramine or salts thereof (e.g. chlorpheniramine maleate), levocabastine or salts thereof (e.g. hydrochloric acid) levocabastine), ketotifen or a salt thereof (eg, ketotifen fumarate), pemirolast potassium, olopatadine or a salt thereof (eg, olopatadine hydrochloride), and the like.
Anti-inflammatory agents: for example, methyl salicylate, glycol salicylate, allantoin, tranexamic acid, lysozyme, lysozyme chloride, indomethacin, pranoprofen, ibuprofen, ibuprofen piconol, ketoprofen, felbinac, bendazac, piroxicam, bufexamac, butyl flufenamate, epsilon- aminocaproic acid, berberine chloride, berberine sulfate, sodium azulene sulfonate, glycyrrhizic acid or a salt thereof (eg, dipotassium glycyrrhizinate, monoammonium glycyrrhizinate) and the like.
Steroid agents: for example, fluticasone propionate, fluticasone furoate, mometasone furoate, beclomethasone propionate, flunisolide and the like.
Decongestants: for example, tetrahydrozoline hydrochloride, tetrahydrozoline nitrate, naphazoline hydrochloride, naphazoline nitrate, epinephrine, epinephrine hydrochloride, ephedrine hydrochloride, phenylephrine hydrochloride, dl-methylephedrine hydrochloride and the like.
Ocular muscle modulating drug: For example, a cholinesterase inhibitor having an active center similar to that of acetylcholine, specifically tropicamide, helenien, atropine sulfate, pilocarpine hydrochloride and the like.
Vitamins: for example, tocopherol acetate, flavin adenine dinucleotide sodium, cyanocobalamin, pyridoxine hydrochloride, panthenol, calcium pantothenate, ascorbic acid, sodium ascorbate and the like.
Amino acids: for example, L-aspartic acid or salts thereof (for example, potassium L-aspartate, sodium L-aspartate, magnesium L-aspartate, calcium L-aspartate, magnesium potassium L-aspartate (L-asparagine an equal mixture of magnesium acid and potassium L-aspartate)), L-arginine, glutamic acid, glycine, alanine, lysine, γ-aminobutyric acid, γ-aminovaleric acid, trimethylglycine and salts thereof, and the like.
Astringents: For example, zinc white, zinc lactate, zinc sulfate and the like.
Others: For example, sulfamethoxazole, sulfisoxazole, sulfisomidine and salts thereof and the like.

In the ophthalmic composition according to the present embodiment, various additives are appropriately selected in accordance with conventional methods according to the application and formulation form, as long as the effects of the present invention are not impaired, and one or more additives are added. may be used in combination and contained in an appropriate amount. Examples of such additives include various additives described in Pharmaceutical Excipient Encyclopedia 2016 (edited by Japan Pharmaceutical Excipients Association). Typical ingredients include the following additives.
Carrier: For example, an aqueous solvent such as water or hydrous ethanol.
Chelating agents: for example, ethylenediaminediacetic acid (EDDA), ethylenediaminetriacetic acid, ethylenediaminetetraacetic acid (EDTA), N-(2-hydroxyethyl)ethylenediaminetriacetic acid (HEDTA), diethylenetriaminepentaacetic acid (DTPA) and the like.
Base: For example, octyldodecanol, titanium oxide, potassium bromide, Plastibase and the like.
pH adjuster: for example, hydrochloric acid, acetic acid, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, triethanolamine, monoethanolamine, diisopropanolamine and the like.
Perfumes or cooling agents: e.g. menthone, camphor, borneol, geraniol, cineol, citronellol, carvone, anethole, eugenol, limonene, linalool, linalyl acetate, thymol, cymene, terpineol, pinene, camphene, isoborneol, fenchen, nerol , myrcene, myrcenol, linalool acetate, lavandulol, eucalyptus oil, bergamot oil, peppermint oil, coolmint oil, spearmint oil, peppermint oil, fennel oil, cinnamon oil, rose oil, camphor oil, etc. These may be d-, l- or dl-isomers.
Thickeners: cellulosic polymer compounds such as methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, sodium carboxymethylcellulose; polyvinylpyrrolidone, polyvinyl alcohol and other polyvinyl polymer compounds; carboxyvinyl polymer; guar gum; hydroxypropylguar gum gum arabic; karaya gum; xanthan gum; agar; alginic acid and its salts (sodium salt, etc.); chitin and its derivatives; chitosan and its derivatives; carrageenan; monosaccharides such as glucose and the like.
Stabilizers: for example, edetic acid, edetate salts (edetate disodium, edetate calcium disodium, edetate trisodium, edetate tetrasodium), sodium formaldehyde sulfoxylate (Rongalite), aluminum monostearate, monostearate glycerin stearate, cyclodextrin, monoethanolamine, dibutylhydroxytoluene, sodium bisulfite, sodium pyrosulfite and the like.
Preservatives: for example, alkylpolyaminoethylglycine quaternary ammonium salts (e.g., benzalkonium chloride, benzethonium chloride, etc.), chlorhexidine gluconate, polydronium chloride, zinc chloride, sodium benzoate, ethanol, chlorobutanol, sorbic acid, Potassium sorbate, sodium dehydroacetate, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, butyl parahydroxybenzoate, oxyquinoline sulfate, phenethyl alcohol, benzyl alcohol, biguanide compounds (specifically, polyhexanide hydrochloride (poly hexamethylenebiguanide), Alexidine, etc.), Glokyl (trade name, manufactured by Rhodia), etc.
Tonicity agents: e.g. potassium chloride, calcium chloride, sodium chloride, magnesium chloride, potassium acetate, sodium acetate, sodium bicarbonate, sodium carbonate, sodium thiosulfate, magnesium sulfate, glycerin, propylene glycol, sodium bisulfite, sodium sulfite etc.
Sugar alcohols: For example, xylitol, sorbitol, mannitol, glycerin and the like. These may be d-, l- or dl-isomers.
Oils: For example, vegetable oils such as sesame oil, castor oil, soybean oil and olive oil; animal oils such as squalane; mineral oils such as liquid paraffin and petrolatum.

When the ophthalmic composition according to the present embodiment contains water, the water content is, for example, based on the total amount of the ophthalmic composition, from the viewpoint of exhibiting the effects of the present invention more remarkably. , 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 90 w/v% or more and 99.2 w/v% or less is more preferred.

The water used in the ophthalmic composition according to this embodiment may be pharmaceutically, pharmacologically (pharmaceutical) or physiologically acceptable. Examples of such water include distilled water, ordinary water, purified water, sterile purified water, water for injection, and distilled water for injection. Their definitions are based on the 17th revision of the Japanese Pharmacopoeia.

The ophthalmic composition according to the present embodiment can be prepared by adding and mixing a desired amount of component (A) and, if necessary, other components to a desired concentration. For example, it can be prepared by dissolving or dispersing these components in purified water, adjusting to a predetermined pH and osmotic pressure, and sterilizing by filtration sterilization or the like.

The ophthalmic composition according to this embodiment can take various formulation forms depending on the purpose. Formulations include, for example, liquids, gels, semi-solids (ointments, etc.) and the like.

The ophthalmic composition according to the present embodiment includes, for example, eye drops (also referred to as eye drops or eye drops. In addition, eye drops include eye drops that can be applied while wearing contact lenses, eye drops for wearing contact lenses, and artificial tears. ), eye washes (also referred to as eye washes or eye washes. Eye washes include eye washes that can be washed while wearing contact lenses and eye washes that are instilled into the eye.), contact lens compositions [ contact lens wetting solution, contact lens care composition (contact lens disinfectant, contact lens preservative, contact lens cleaning agent, contact lens cleaning preservative), contact lens packaging solution, etc.]. The term "contact lens" includes hard contact lenses and soft contact lenses (both ionic and non-ionic, including both silicone hydrogel contact lenses and non-silicone hydrogel contact lenses).

Since the ophthalmic composition according to the present embodiment has a protective effect on the cornea, it can be suitably used as an ophthalmic composition for protecting the cornea. In addition, the ophthalmic composition according to the present embodiment is used to prevent inflammation (preferably inflammation caused by dry eyes and eye fatigue, more preferably dry eyes and eye fatigue caused by use of digital devices and aging) Inflammation), wearing of contact lenses, etc., and since it has an effect of repairing or recovering the cornea, it can be suitably used as an ophthalmic composition for repairing or recovering corneal damage. In addition, since the ophthalmic composition according to the present embodiment increases transepithelial electrical resistance and enhances the expression of occludin, it is used for improving the barrier function of the cornea, suppressing foreign body sensation, or preventing infectious diseases or allergies. It can also be suitably used as an ophthalmic composition. In addition, since the ophthalmic composition according to the present embodiment increases the expression of membrane-type mucin, it is also suitable as an ophthalmic composition for improving the tear retention function of the cornea, stabilizing tear fluid, or suppressing foreign body sensation. can be used for In addition, since the ophthalmic composition according to the present embodiment has a corneal protective action and a corneal repairing or recovering action, it causes eye fatigue, blurred vision, and eye discomfort (especially when wearing contact lenses). Discomfort) and other symptoms can be suitably used. In addition, since the ophthalmic composition according to the present embodiment increases transepithelial electrical resistance and enhances the expression of occludin, it can be suitably used for symptoms such as itchy eyes, watery eyes, and foreign body sensation. and can be suitably used for the prevention of eye diseases. In addition, since the ophthalmic composition according to the present embodiment improves the function of retaining tear fluid, it can be suitably used for symptoms such as dry eyes, foreign body sensation, and tired eyes due to dry eyes.

The ophthalmic composition according to this embodiment is preferably an eye drop (including an eye drop that can be instilled while wearing contact lenses), since the effects of the present invention can be exhibited more remarkably. When the ophthalmic composition according to the present embodiment is an eye drop, the usage and dosage are not particularly limited as long as they are effective and have few side effects. In the case of children above, the method of using 1 to 3 drops, 1 to 2 drops, or 2 to 3 drops 2 to 4 times a day, or 5 to 6 times a day is exemplified, and the effect of the present invention is demonstrated. From the viewpoint of achieving a more pronounced effect, a method of applying 1 to 2 drops per time, 3 to 4 times a day, is preferred, and a method of applying 1 to 2 drops per time, 4 times a day is more preferred. The ophthalmic composition according to the present embodiment can be used by those who have subjective symptoms such as dry eyes, eye fatigue, blurred vision, and eye discomfort (especially eye discomfort when wearing contact lenses). By using it for a certain period of time (preferably 1 week or more, more preferably 2 weeks or more), the frequency of occurrence of these subjective symptoms can be suppressed.

The ophthalmic composition according to this embodiment is provided in an arbitrary container. The container for containing the ophthalmic composition according to this embodiment is not particularly limited, and may be made of glass or plastic, for example. It is preferably made of plastic. Examples of plastics include polyethylene terephthalate, polyarylate, polyethylene naphthalate, polycarbonate, polyethylene, polypropylene, polyimide, copolymers of these monomers, and mixtures of two or more thereof. Preferred is polyethylene terephthalate. Further, the container containing the ophthalmic composition according to the present embodiment may be a transparent container in which the inside of the container is visible, or an opaque container in which the inside of the container is difficult to be seen. A transparent container is preferred. Here, the "transparent container" includes both a colorless transparent container and a colored transparent container.

A nozzle may be attached to the container containing the ophthalmic composition according to the present embodiment. The material of the nozzle is not particularly limited, and may be, for example, glass or plastic. It is preferably made of plastic. Examples of plastics include polybutylene terephthalate, polyethylene, polypropylene, polyethylene terephthalate, polyethylene naphthalate, copolymers of monomers constituting these, and mixtures of two or more of these. From the viewpoint of further enhancing the effects of the present invention, the nozzle material is preferably polypropylene, polyethylene, polyethylene terephthalate, polybutylene terephthalate, or polyethylene naphthalate, and more preferably polyethylene.

The container containing the ophthalmic composition according to the present embodiment may be of a multi-dose type that accommodates multiple doses, or may be of a unit dose type that accommodates a single dose.

The ophthalmic composition according to the present 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. More preferably, it is packed in a container of 6 to 16 mL, and even more preferably packed in a container with an internal volume of 10 to 15 mL. Also, it may be filled in a container with an internal volume of 0.1 to 3 mL, or may be filled in a container with an internal volume of 0.2 to 1 mL.

As described above, the ophthalmic composition according to the present embodiment has a corneal protective effect and an effect of repairing or recovering the cornea damaged by inflammation or the like. (2) chondroitin sulfate with a weight average molecular weight of 40,000 to 70,000; (3) chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and a salt thereof in an ophthalmic composition; and (4) chondroitin having a weight average molecular weight of 40,000 to 70,000 in the ophthalmic composition. Provided is a method for imparting a corneal injury repairing or injury recovery action to the ophthalmic composition, comprising containing at least one selected from the group consisting of sulfuric acid and salts thereof.

The present invention will be specifically described below based on test examples, but the present invention is not limited to these. The sodium chondroitin sulfate used in the test examples below is as follows, and the sodium chondroitin sulfate having a weight-average molecular weight of about 56,000 is derived from sharks.
Chondroitin sulfate sodium weight average molecular weight about 56000: Seikagaku Corporation; grade NK
Weight average molecular weight about 25000: Maruha Nichiro Co., Ltd.;

[Test Example 1: Corneal protective effect on normal corneal epithelial cell line (transepithelial electrical resistance)]
Corneal epithelial cell lines were cultured to confluence on cell culture inserts coated with laminin 511E8 fragment (0.5 μg/cm ² ) (Nippi). Differentiation medium [2% B27 Supplement (Life Technologies), 1% penicillin-streptomycin solution (Life Technologies), 20 ng/mL KGF (Keratinocyte Growth Factor) (Wako), 10 μM Y- 27632 (Wako) was included DMEM/F-12 (2:1 (v/v)) medium (Life Technologies)] was used. Subsequently, the cells were starved by overnight culture in a basal medium [DMEM/F-12 (2:1 (v/v) medium containing 1% penicillin-streptomycin solution)]. After that, sodium chondroitin sulfate (Seikagaku Corporation) with a weight average molecular weight of about 56000 was added to the basal medium so that the concentration was 0.5%. Transepithelial electrical resistance (TER) was measured. A basal medium to which sodium chondroitin sulfate was not added was used as a control. The results are shown in Table 1 and FIG. Here, it is evaluated that the higher the transepithelial electrical resistance, the stronger the corneal protective effect.

Sodium chondroitin sulfate having a weight-average molecular weight of about 56,000 was confirmed to increase transepithelial electrical resistance, strengthen the corneal barrier function, and strongly protect the cornea.

[Test Example 2: Corneal damage recovery effect on corneal epithelial cell line during inflammation (transepithelial electrical resistance)]
Corneal epithelial cell lines were cultured to confluence on cell culture inserts coated with laminin 511E8 fragment (0.5 μg/cm ² ). Differentiation medium was used for culture. Subsequently, the cells were cultured overnight in the same basal medium as in Test Example 1 to starve the cells. Thereafter, TNF (tumor necrosis factor) (PEPROTECH) was added (10 ng/mL) as an inflammatory factor to the basal medium, and then sodium chondroitin sulfate with a weight average molecular weight of about 56,000 or sodium chondroitin sulfate with a weight average molecular weight of about 25,000 was added. After each addition to a concentration of 0.5%, culture was performed for 48 hours, and transepithelial electrical resistance (TER) was measured using Millicell-ERS2. The culture condition in which neither TNF nor sodium chondroitin sulfate was added was used as a negative control, and the culture condition in which TNF was added but no sodium chondroitin sulfate was added was used as a positive control. The results are shown in Table 2 and FIG. Here, it is evaluated that the higher the transepithelial electrical resistance, the stronger the inhibitory effect on corneal damage.

It was confirmed that sodium chondroitin sulfate with a weight average molecular weight of about 56,000 has a higher transepithelial electrical resistance than sodium chondroitin sulfate with a weight average molecular weight of about 25,000, and has a high inhibitory effect on corneal damage. Therefore, chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and a salt thereof can be applied to the recovery of the cornea damaged by inflammation.

[Test Example 3: Corneal protective action (gene expression) in normal corneal epithelial cell line]
Corneal epithelial cell lines were cultured on cell culture dishes coated with laminin 511E8 fragment (0.5 μg/cm ² ) until confluent. The same differentiation medium as in Test Example 1 was used for culture. Subsequently, the cells were cultured overnight in the same basal medium as in Test Example 1 to starve the cells. Thereafter, sodium chondroitin sulfate having a weight average molecular weight of about 56,000 or sodium chondroitin sulfate having a weight average molecular weight of about 25,000 was added to the basal medium to a concentration of 0.5%, and after culturing for 24 hours, Maxwell RSC simplyRNA tissue was added. Cells were harvested using Kit (Promega), total RNA was extracted, and changes in gene expression levels of occludin (OCLN) relative to GAPDH used as an endogenous control were measured. A basal medium to which sodium chondroitin sulfate was not added was used as a control.
For extraction of total RNA, Maxwell RSC simplyRNA Tissue Kit (Promega) was used. Reverse transcription PCR was performed using ReverTra Ace qPCR RT Master Mix (TOYOBO) to prepare cDNA. Using the Cybergreen method, mRNA expression levels of GAPDH, OCLN and MUC16 were evaluated by quantitative real-time PCR using QuantStudio 7 Flex (Thermo Fisher Scientific). After quantitative real-time PCR, the CT value of each sample was calculated by automatic analysis of QuantStudio™ Design & Analysis Software.
The ΔCT value of the target factor in each sample was calculated from equation (1).
Formula (1): ΔCT value = (CT value of target factor) - (CT value of GAPDH)
The average value of the ΔCT values of the samples under static culture conditions was calculated, and the ΔΔCT value of each sample was calculated from Equation (2).
Formula (2): ΔΔCT value = (ΔCT value of target factor) - (average value of ΔCT values of samples under static culture conditions)
The relative mRNA expression level of each sample with respect to static culture conditions was calculated from the formula (3).
Formula (3): Relative mRNA expression level of each sample=2-ΔΔCT value The results are shown in Table 3 and FIG. Here, it is evaluated that the higher the gene expression of occludin, the stronger the corneal protective effect.

Sodium chondroitin sulfate with a weight-average molecular weight of about 56,000 increases the gene expression level of occludin and has stronger corneal barrier function-strengthening and corneal-protecting effects than sodium chondroitin sulfate with a weight-average molecular weight of about 25,000. was confirmed.

[Test Example 4: Corneal damage recovery effect on corneal epithelial cell line during inflammation (gene expression)]
Corneal epithelial cell lines were cultured on cell culture dishes coated with laminin 511E8 fragment (0.5 μg/cm ² ) until confluent. The same differentiation medium as in Test Example 1 was used for culture. Subsequently, the cells were cultured overnight in the same basal medium as in Test Example 1 to starve the cells. Then, TNF (10 ng/mL) was added to the basal medium as an inflammatory factor, and then sodium chondroitin sulfate with a weight average molecular weight of about 56,000 or chondroitin sulfate with a weight average molecular weight of about 25,000 was added to the basal medium. Sodium was added to each to a concentration of 0.5%, and after culturing for 24 hours, cells were collected using Maxwell RSC simply RNA Tissue Kit (Promega). After that, RT-qPCR was performed to measure changes in gene expression levels of occludin (OCLN) and membranous mucin (MUC16) relative to GAPDH used as an endogenous control. Measurement conditions conform to Test Example 3. The culture condition in which neither TNF nor sodium chondroitin sulfate was added was used as a negative control, and the culture condition in which TNF was added but no sodium chondroitin sulfate was added was used as a positive control. The results are shown in Table 4, Figures 4 and 5. Here, it is evaluated that the greater the gene expression of occludin and membranous mucin, the stronger the inhibitory effect on corneal damage.

Sodium chondroitin sulfate with a weight-average molecular weight of about 56,000 increased the gene expression levels of occludin and membranous mucin compared to sodium chondroitin sulfate with a weight-average molecular weight of about 25,000, confirming that it has a strong inhibitory effect on corneal disorders. rice field. Therefore, chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and a salt thereof can be applied to the recovery of the cornea damaged by inflammation. In addition, since the gene expression level of membranous mucin increased, chondroitin sulfate and its salts with a weight average molecular weight of 40,000 to 70,000 can be applied to improve the corneal tear retention function and stabilize tear fluid. .

[Test Example 5: Corneal injury recovery effect on corneal epithelial cell line during inflammation (immunostaining)]
Corneal epithelial cell lines were cultured on cell culture dishes coated with laminin 511E8 fragment (0.5 μg/cm ² ) until confluent. The same differentiation medium as in Test Example 1 was used for culture. Subsequently, the cells were cultured overnight in the same basal medium as in Test Example 1 to starve the cells. Thereafter, TNF (10 ng/mL) was added to the basal medium as an inflammatory factor, and then sodium chondroitin sulfate with a weight average molecular weight of about 56,000 or sodium chondroitin sulfate with a weight average molecular weight of about 25,000 was added to the basal medium at a concentration of 0.5. After culturing for 48 hours, the cells were fixed with 4% paraformaldehyde/phosphate buffer (Wako). Subsequently, after blocking (room temperature, 1 hour) with 5% NST [5% Normal Donkey Serum (Jackson Immuno Research), TBS (Takara) containing 0.3% Triton X-100 (Sigma)], anti-occludin (OCLN ) antibody (1:200; ab216327, abcam) and an anti-membrane type mucin (MUC16) antibody (1:1000; ab1107, abcam) were used to perform a primary antibody reaction (4° C., overnight). After washing twice with TBS for 5 minutes, treatment with secondary antibody (1:200; Thermo Fisher Scientific) labeled with Alexa Fluor 488 or Alexa Fluor 568 and Hoechst 33342 (1:100; B2261-25MG, Merck) (room temperature). , 1 hour), the cells were observed under a fluorescence microscope. The culture condition in which neither TNF nor sodium chondroitin sulfate was added was used as a negative control, and the culture condition in which TNF was added but no sodium chondroitin sulfate was added was used as a positive control. FIG. 6 shows a photograph of immunostaining for occludin, and FIG. 7 shows a photograph of immunostaining for membranous mucin. Here, in FIG. 6 and FIG. 7, (a) is a negative control, (b) is a positive control, (c) is sodium chondroitin sulfate with a weight average molecular weight of about 56000, and (d) is a weight average molecular weight. Sodium chondroitin sulfate with a weight of about 25,000 was added, and the expression of occludin and membranous mucin was promoted as the area of the white portion in the photograph increased, and it was evaluated that the inhibitory effect on corneal damage was stronger.

6 and 7, sodium chondroitin sulfate with a weight average molecular weight of about 56,000 promotes the expression of both occludin and membranous mucin compared to sodium chondroitin sulfate with a weight average molecular weight of about 25,000. It could be confirmed. Therefore, chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and a salt thereof can be applied to the recovery of the cornea damaged by inflammation. In addition, since the expression of membranous mucin was promoted, chondroitin sulfate and its salts having a weight average molecular weight of 40,000 to 70,000 can be applied to improve the corneal tear fluid retention function and stabilize tear fluid.

[Formulation example]
Eye drops are prepared by a conventional method with the formulations shown in Tables 5 to 11 below. In addition, the unit of each component amount in Tables 5 to 11 below is w/v % except for those specified in the table.

Claims (2)

An ophthalmic composition for protecting the cornea, containing at least one selected from the group consisting of chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and salts thereof. An ophthalmic composition for repairing or recovering corneal damage, containing at least one selected from the group consisting of chondroitin sulfate having a weight average molecular weight of 40,000 to 70,000 and salts thereof.

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