JP2021183606A - Liquid formulation containing brimonidine - Google Patents

Abstract

To provide ophthalmic liquid preparations containing brimonidine or a salt thereof.SOLUTION: We have found that an ophthalmic liquid preparation containing a low concentration of brimonidine supplemented with chondroitin sulfate ester and/or a salt thereof has high transferability of conjunctival tissue and reduces cytotoxicity due to brimonidine, and provide an ophthalmic liquid preparation containing low-concentration brimonidine and chondroitin sulfate ester and/or a salt thereof.SELECTED DRAWING: Figure 2

Description

The present invention relates to a liquid preparation containing brimonidine or a salt thereof.

Brimonidine and its salts are known as α-2 adrenergic receptor agonists. The human eye has many α-2 adrenergic receptors (hereinafter, may be abbreviated as α-2 receptors), and the agonist of the α-2 receptor is an intraocular humor and an actuarial membrane. It has the effect of lowering intraocular pressure by promoting the outflow of aqueous humor through the scleral outflow tract. Based on this effect, α-2 receptor agonists have conventionally been used for the treatment of glaucoma and ocular hypertension. In addition, the α-2 receptor agonist has an action of causing a reduction in the lumen size of the α-2 receptor concentrated arteriole, particularly the terminal arteriole. By this action, vasoconstriction can occur, the redness of the eye can be reduced, the whiteness can be increased, and the aesthetic appearance of the eye can be improved (Patent Document 1: Japanese Patent No. 5671459; Patent Document 2: Patent Document 2: Patent Document 2: Patent Document 2: Patent Document 2: Patent Document 2: Patent Document 2: Patent Document 2: Patent Document 2: No. 5738890).

For a pharmaceutical product in which brimonidine and / or a salt thereof and timolol and / or a salt thereof are used in combination, a pharmaceutical formulation technique focusing on the pharmaceutical stability is also being studied. For example, Patent Document 3 (Japanese Unexamined Patent Publication No. 2019-104727) discloses an aqueous liquid preparation containing brimonidine and / or a salt thereof, a water-soluble polymer, and chlorhexidine gluconate. In this document, examples of the water-soluble polymer include carboxymethyl cellulose and / or a salt thereof, polyvinyl alcohol, polyvinylpyrrolidone, hydroxypropylmethyl cellulose, and hyaluronic acid, and the combination of these components has high viscosity stability. It is disclosed that deterioration is suppressed. Further, Patent Document 4 (Japanese Unexamined Patent Publication No. 2019-178083) discloses an aqueous solution for eye drops having a pH of less than 7.4 containing brimonidine or a salt thereof and a water-soluble polymer. In this document, polysaccharides, cellulose polymers, and synthetic polymers are mentioned as water-soluble polymers, and when sodium carboxymethyl cellulose, hydroxypropylmethyl cellulose, and polyvinylpyrrolidone are used, precipitation of insoluble matter is suppressed and storage stability is achieved. Is disclosed to be excellent.

Japanese Patent No. 5671459 Japanese Patent No. 5738890 Japanese Unexamined Patent Publication No. 2019-104727 Japanese Unexamined Patent Publication No. 2019-178083

It is an object of the present invention to provide an ophthalmic liquid preparation having more excellent efficacy in formulating brimonidine or a salt thereof as an ophthalmic liquid preparation for alleviating or suppressing red eye.

The present inventors have conducted diligent studies for the purpose of providing an ophthalmic liquid preparation containing a low concentration of brimonidine, and found that a low concentration brimonidine is contained by incorporating a chondroitin sulfate ester and / or a salt thereof. It has been found that the cytotoxicity of liquid formulations is reduced. It was also found that an ophthalmic liquid preparation containing a low concentration of brimonidine to which a chondroitin sulfate ester and / or a salt thereof was added has high conjunctival tissue transferability.

As an embodiment of the present invention, the following liquid preparation for ophthalmology is provided.
[1] An ophthalmic liquid preparation containing 0.01 to 0.05 w / v% of brimonidine and / or a salt thereof, and a chondroitin sulfate ester and / or a salt thereof.
[2] The liquid preparation for ophthalmology according to item 1, wherein the salt of chondroitin sulfate is a sodium salt.
[3] The liquid preparation for ophthalmology according to item 1 or 2, wherein the content of chondroitin sulfate ester and / or a salt thereof is 0.05 to 0.5 w / v%.

Further, as an embodiment of the present invention, the following method for reducing cytotoxicity is provided.
[4] Brimonidine in eye drop administration, which comprises blending a chondroitin sulfate ester and / or a salt thereof with an ophthalmic liquid preparation containing 0.01 to 0.05 w / v% of brimonidine and / or a salt thereof. And / or a method for reducing cytotoxicity of a salt thereof.

Further, as an embodiment of the present invention, the following cytotoxicity reducing agent is provided.
[5] Brimonidine in eye drop administration, which comprises blending a chondroitin sulfate ester and / or a salt thereof with an ophthalmic liquid preparation containing 0.01 to 0.05 w / v% of brimonidine and / or a salt thereof. And / or a cytotoxicity-reducing agent of a salt thereof.

Further, as an embodiment of the present invention, the following method for promoting conjunctival migration is provided.
[6] Brimonidine in eye drop administration, which comprises blending a chondroitin sulfate ester and / or a salt thereof with an ophthalmic liquid preparation containing 0.01 to 0.05 w / v% of brimonidine and / or a salt thereof. And / or a method for promoting conjunctival transfer of a salt thereof.

Further, as an embodiment of the present invention, the following conjunctival transfer promoter is provided.
[7] Brimonidine in eye drop administration, which comprises blending a chondroitin sulfate ester and / or a salt thereof with an ophthalmic liquid preparation containing 0.01 to 0.05 w / v% of brimonidine and / or a salt thereof. And / or a conjunctival transfer promoter of a salt thereof.

According to the present invention, an ophthalmic liquid preparation containing a low concentration of brimonidine and chondroitin sulfate ester and / or a salt thereof has at least one effect of reducing the cytotoxicity of brimonidine and improving the conjunctival tissue transferability of brimonidine. Demonstrate.

FIG. 1 is a graph showing the effects of brimonidine tartrate and sodium chondroitin sulfate on cultured cells. The vertical axis shows the absorbance of the well after color development by the Cell counting kit, and the higher the absorbance, the higher the number of living cells. Low control refers to the group that does not contain cells, and high control refers to the group that uses cell culture medium instead of the test solution containing the drug. Although the absorbance decreased in the group to which brimonidine tartrate was added (BRN alone), the group to which sodium chondroitin sulfate was added (CS alone) and the group to which brimonidine tartrate and sodium chondroitin sulfate were added (CS + BRN) had high control. It showed the same degree of absorbance as. FIG. 2 is a graph showing the effects of brimonidine tartrate and sodium chondroitin sulfate on cultured cells. The vertical axis shows the absorbance of the well after color development by the Cell counting kit, and the higher the absorbance, the higher the number of living cells. Low control refers to the group that does not contain cells, and high control refers to the group that uses cell culture medium instead of the test solution containing the drug. Although the absorbance decreased in the group to which brimonidine tartrate was added (BRN 0.025% alone), the group to which brimonidine tartrate and 0.1 w / v% sodium chondroitin sulfate were added (CS 0.1% + BRN 0.025%), The group to which brimonidine tartrate and 0.3 w / v% sodium chondroitin sulfate were added (CS 0.3% + BRN 0.025%), the group to which brimonidine tartrate and 0.5% sodium chondroitin sulfate were added (CS 0.5%). At + BRN 0.025%), the absorbance was similar to that of High Control. FIG. 3 is a graph showing the effects of brimonidine tartrate and sodium chondroitin sulfate on cultured cells. The vertical axis shows the absorbance of the well after color development by the Cell counting kit, and the higher the absorbance, the higher the number of living cells. Low control refers to the group that does not contain cells, and high control refers to the group that uses cell culture medium instead of the test solution containing the drug. The group to which only 0.5 w / v% chondroitin sulfate sodium was added (CS 0.5% alone), the group to which 0.025 w / v% brimonidine tartrate and 0.5 w / v% chondroitin sulfate sodium were added (CS0. 5% + BRN 0.025%), 0.05w / v% brimonidine tartrate and 0.5w / v% sodium chondroitin sulfate (CS 0.5% + BRN 0.05%) were comparable to high control. The absorbance was shown.

It is understood that the terms used herein are used in the meaning commonly used in the art unless otherwise noted. Accordingly, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The numerical range specified herein shall include its lower and upper limits.

(Definition)
As used herein, "brimonidine" is referred to as IUPAC name: 5-bromo-N- (4,5-dihydro-1H-imidazol-2-yl) quinoxaline-6-amine (5-Bromo-N- (4,5)). -Dihydro-1H-imidazol-2-yl) refers to a compound of quinoxaline-6-amine). Further, in the present specification, the concentration of brimonidine and / or a salt thereof is the concentration converted to brimonidine tartrate unless otherwise specified.

As used herein, the term "low concentration brimonidine" refers to brimonidine having a concentration of 0.05 w / v% or less.

As used herein, the "liquid preparation for ophthalmology" is an aqueous liquid preparation based on water.

As used herein, the term "cytotoxicity" refers to the property of brimonidine causing cell death, dysfunction or inhibition of proliferation of cells of ocular tissue.

In the present specification, "reduction of cytotoxicity" or "reduction of cytotoxicity" means suppressing the influence of cytotoxicity and continuing to maintain the original function of the cell.

As used herein, the term "cytotoxicity reducing method" means a method used to suppress the effect of brimonidine on cytotoxicity on ocular tissues and to maintain the original function of cells.

As used herein, the term "cytotoxicity reducing agent" means an agent formulated to suppress the effect of brimonidine on cytotoxicity on ocular tissues and to maintain the original function of cells.

As used herein, the term "conjunctival transferability" is an index indicating the ease with which brimonidine in an ophthalmic liquid preparation administered by eye drops can be transferred to the conjunctiva, which is the site of action.

As used herein, the term "method for promoting conjunctival transfer" means a method performed to increase the amount of brimonidine transferred to the conjunctiva, which is a site of action, in an ophthalmic liquid preparation administered by eye drops.

As used herein, the term "conjunctival transfer promoter" means an agent formulated to increase the amount of brimonidine transferred to the conjunctiva, which is the site of action, in an ophthalmic liquid preparation administered by eye drops.

(Explanation of Preferred Embodiment)
Hereinafter, preferred embodiments of the present invention will be described. It is understood that the embodiments provided below are provided for a better understanding of the invention and the scope of the invention should not be limited to the following description. Therefore, it is clear that a person skilled in the art can appropriately make modifications within the scope of the present invention in consideration of the description in the present specification. It is also understood that the following embodiments may be used alone or in combination thereof.

In the present invention, any salt can be mentioned as a pharmaceutically acceptable salt of brimonidine. Pharmaceutically acceptable salts of brimonidine include hydrochloride, sulfate, phosphate, acetate, citrate, oxalate, malonate, salicylate, malate, fumarate, succinate, Examples thereof include ascorbate, maleate, methanesulfonate, tartrate and other inorganic carboxylates well known to those skilled in the art, preferably tartrate. In addition, in this specification, brimonidine and / or a salt thereof may be referred to as "BRN".

In the present invention, the low concentration brimonidine refers to brimonidine having a concentration of 0.05 w / v% or less. As an example, the upper limit of the concentration may be 0.04 w / v% or 0.03 w / v% from the viewpoint of preventing side effects. The lower limit of the concentration is not limited as long as brimonidine is contained, but 0.01 w / v% may be used. As an example, in view of the effect of this drug, 0.015 w / v% or 0.02 w / v. % May be used.

In the present invention, the chondroitin sulfate ester and / or the salt thereof contained in the liquid preparation containing brimonidine or a salt thereof is a kind of glycosaminoglycan found in a living body. The chondroitin sulfate ester has a structure in which the disaccharides of D-glucuronic acid (GlcA) and N-acetyl-D-galactosamine (GalNAc) are repeated, and the 4-position and / or 6-position of GalNAc has a sulfate group. , The structure may differ depending on the species of origin. In the chondroitin sulfate ester and / or its salt of the present invention, the disaccharides of D-glucuronic acid (GlcA) and N-acetyl-D-galactosamine (GalNAc) are repeated, and the 4-position and / or 6-position of GalNAc is a sulfate group. It is not limited to those having the above, and includes all commercially available chondroitin sulfate esters. The chondroitin sulfate ester may exist in the form of any salt, and may exist as a sodium salt or a potassium salt as an example. Chondroitin sulfate ester and / or a salt thereof is added to eye drops for the purpose of protecting the corneal surface layer based on its water retention effect. In the present invention, brimonidine or a salt thereof exerts an action of reducing cytotoxicity. This action can also be referred to as a cytoprotective action. Further, in the present invention, the conjunctival transferability of brimonidine is enhanced by blending chondroitin sulfate ester or a salt thereof. The chondroitin sulfate ester and / or a salt thereof may be blended at 0.005 w / v% to 5 w / v%, more preferably 0.05 to 0.5 w / v%. From the viewpoint of reducing cytotoxicity, the lower limit of the concentration may be 0.01 w / v%, 0.05 w / v%, or 0.1 w / v% as an example. From the viewpoint of blending with eye drops, the upper limit of the concentration may be 3 w / v%, 1 w / v% or 0.5 w / v% as an example. In addition, in this specification, a chondroitin sulfate ester and / or a salt thereof may be referred to as "CS". As an example of an embodiment, the liquid preparation of the present invention containing brimonidine or a salt thereof and a chondroitin sulfate ester and / or a salt thereof may be characterized by not containing chlorhexidine gluconic acid or a salt thereof.

The conjunctiva is a membrane that covers the sclera, which is the white of the eye, and also covers the inside of the eyelid, and is mainly composed of conjunctival epithelial cells. The epithelial layer of the conjunctiva contains blood vessels, fibrous tissue, and lymphatic vessels. It is in contact with the cornea at the boundary between the white and black eyes, and the cornea and conjunctiva form the outermost layer of the eye exposed to the outside world. Since the conjunctiva is exposed to the outside world, it is easily attacked by bacteria and viruses, and inflammation is likely to occur. Even when inflammation does not occur, lack of sleep and overuse of the eyes increase blood flow to supply oxygen and nutrients to the eyes, resulting in redness. In the present invention, in order for brimonidine to exert the effect of reducing or whitening the redness of the eye, it is necessary that the active ingredient of the pharmaceutical product administered by eye drops reaches the capillaries in the conjunctiva and acts.

The conjunctival transferability is an index showing the ease of transfer of the active ingredient (brimonidine or a salt thereof in the present invention) of the ophthalmic liquid preparation administered by eye drops to the conjunctiva, which is the site of action. Corneal epithelial cells and conjunctival epithelial cells each form tight junctions between cells to form a hydrophobic membrane resulting from the cell membrane. This limits the permeation of water-soluble drugs through the cornea and conjunctiva. The transferability of the instilled drug through the cornea and conjunctiva is determined by measuring the drug concentration in the aqueous humor and the drug concentration in the conjunctival tissue after instillation, but it is difficult to measure in humans and animals. Analysis with a predictive model through experiments is being performed. As a simple method for measuring intraocular transferability, a method based on the octanol / water partition coefficient is known. The octanol / water partition coefficient can be used to evaluate not only intraocular transferability but also conjunctival transferability.

In the present invention, the liquid preparation for ophthalmology is an aqueous liquid preparation mainly based on water, but may further contain any liquid base that can be used as an eye drop. The liquid preparation for ophthalmology of the present invention is prepared so as to have a pH and an osmotic pressure acceptable as an eye drop. The pH of the liquid preparation for ophthalmology can be adjusted to 5.0 to 9.0 using a pH adjuster, for example 5.5 to 8.5. As an example, it can be adjusted to 6.0 to 8.0. As an example of the osmotic pressure ratio of the liquid preparation for ophthalmology, the amount of the component added to the liquid preparation for ophthalmology is adjusted so as to be preferably 0.5 to 2.5, more preferably 0.7 to 1.5.

The liquid preparation for ophthalmology of the present invention is preferably an eye drop. The liquid preparation for ophthalmology of the present invention may be a liquid preparation that relieves or suppresses red eye. Relieving or suppressing red eye refers to increasing the whiteness of the white part of the eye, and can also be referred to as eye whitening.

The liquid preparation for ophthalmology of the present invention can contain any component that can be used for eye drops as long as the effect of the present invention is not impaired. In addition to brimonidine tartrate, which is the active ingredient of the present invention, any active ingredient and an ingredient such as an additive may be contained. Such ingredients include decongestants, focus control function improvers, anti-inflammatory / astringent agents, antihistamines, vitamins, nutritional ingredients, sulfa agents, preservatives, pH regulators, isotonic agents, thickeners, antioxidants. Agents, solubilizers, stabilizers, surfactants, fragrances or refreshing agents, etc. may be mentioned, but are not intended to be limited thereto. Only one kind of these active ingredients and additives may be used from each category, or a plurality of kinds may be used in combination.

As the decongestant, for example, epinephrine, ephedrine, tetrahydrozoline, naphazoline, phenylephrine, methylephedrine, or salts thereof may be used.

As the focus adjusting function improving agent, neostigmine methylsulfate can be used.

As the anti-inflammatory / astringent agent, for example, ε-aminocaproic acid, allantin, velverine or a salt thereof, azulene sulfonic acid or a salt thereof, glycyrrhizinic acid or a salt thereof, zinc sulfate, zinc lactate, lysoteam chloride and the like can be used.

As the antihistamine, diphenhydramine hydrochloride and chlorpheniramine maleate can be used.

As the nutritional component, amino acids or salts thereof may be used. As amino acids or salts thereof, amino acids are not only amino acids but also substances having a sulfate group instead of the carboxyl group of the amino acid, for example, taurine. Examples of amino acids include glycine, alanine, methionine, valine, threonine, glutamine, glutamic acid, aspartin, aspartic acid, cysteine, histidine, isoleucine, leucine, lysine, phenylalanine, tryptophan, arginine, proline, tyrosine and serine. .. Amino acids preferably include aspartic acid, methionine and glycine. The amino acids other than glycine may be L-form amino acids, D-form amino acids, or DL-form amino acids.

Vitamins are a general term for organic compounds other than carbohydrates, proteins, and lipids that cannot synthesize a sufficient amount of nutrients necessary for the survival or growth of an organism in the body of the organism. Vitamins are roughly classified into water-soluble vitamins and fat-soluble vitamins. Examples of water-soluble vitamins include vitamin Bs and vitamin C (ascorbic acid). Examples of fat-soluble vitamins include vitamin A, vitamin D, vitamin E, and vitamin K. The over-the-counter drug manufacturing (import) approval standard stipulates vitamins to be blended in eye drops, and from this viewpoint, vitamin A, vitamin B, and vitamin E are particularly preferable.

As vitamin A, retinol and related substances thereof can be used. Retinol-related substances include retinal, retinoic acid, retinol palmitate and other retinoids such as isotretinoin, alitretinoin, aitretinoin, etretinate, adaparene, tazarotene, bexarotene and the like. Retinyl palmitate and retinyl acetate are preferable from the viewpoint of blending as eye drops. Since vitamins A act on epithelial cells and induce proliferation, they can be added to eye drops for the purpose of protecting the cornea and conjunctiva. It can also be added to eye drops for the treatment of ophthalmic diseases such as night blindness, dry conjunctival disease, dry corneal disease, and keratomalacia.

As the B vitamins, vitamin B1 (thiamin and the like), vitamin B2, vitamin B3 (niacin and the like), vitamin B5, vitamin B6, vitamin B7 (biotin and the like), vitamin B9 (folic acid and the like) and vitamin B12 can be used. Vitamins include derivative provitamins and pharmaceutically acceptable salts. Among the B vitamins, vitamins B2, B5, B6 and B12 are particularly preferable from the viewpoint of being blended in eye drops.

As vitamin B2, riboflavin, riboflavin phosphate, riboflavin butyrate, riboflavin acetate, flavin adenine dinucleotide, flavin mononucleotide, pharmaceutically acceptable salts thereof and the like can be used. Examples of the salt include sodium salt and potassium salt. Flavin adenine dinucliotide sodium is preferable from the viewpoint of being compounded as an eye drop. Vitamin B2 is directly involved in oxidative reduction, and when used as an eye drop, promotes enzymatic respiratory metabolism of corneal and conjunctival cells, and has a protective effect on the cornea and conjunctiva. Vitamin B2 can also be added to eye drops for the treatment of keratitis presumed to be associated with vitamin B2 deficiency or metabolic disorders.

As vitamin B5, panthenol, pantothenic acid or derivatives thereof or salts thereof can be used. As an example, in addition to panthenol and pantothenic acid, as derivatives or salts thereof, pantethine, pantetheine, pantotheneal alcohol, pantotheneyl ethyl ether, pantetheine pantothenic acid alcohol, calcium pantothenate, sodium pantothenate and the like can be mentioned. From the viewpoint of being used as eye drops, panthenol, calcium pantothenate, and sodium pantothenate are preferable as vitamin B5.

As vitamin B6, pyridoxal, pyridoxamine, pyridoxine or pharmaceutically acceptable salts thereof can be used. Pyridoxine hydrochloride is preferable from the viewpoint of being used as an eye drop. Vitamin B6 is involved in protein metabolism as a coenzyme of amino acid decarboxylase and transaminase in vivo, and can be added to eye drops to suppress eye fatigue.

Vitamin B12 is a compound having a structure in which cobalt is coordinated to the choline ring, and specific examples thereof include cyanocobalamin, mecobalamin (methylcobalamin), hydroxocobalamin, adenosylcobalamin, hydroxocobalamin hydrochloride, and hydroxocobalamin acetate. Vitamin B12 can be added to eye drops for pharmacological actions such as improvement of tired eyes and eyestrain.

As vitamin Es, tocopherol, tocotrienol, tocophersolan, or derivatives thereof can be used. The tocopherol and tocotrienol may be any of α-, β-, γ-, and δ-, and may be any of d-form and dl-form. From the viewpoint of being used as an eye drop, d-α-tocopherol acetate can be mentioned as an example.

Vitamin C includes ascorbic acid or a salt thereof. Vitamin Ds include vitamin D2 (ergosterol, ergocalciferol), D3 (7-dehydrocholesterol), previtamin D3 (cholecalciferol, 25-hydroxycholecalciferol, calcitriol (1,25-dihydroxycholecalcifer)). Ferrol), calcitronic acid), vitamin D4 (dihydroergocalciferol), vitamin D5 (dihydrotaxterol, calcipotriol, tacalcitol, paricalcitol) and the like. Examples of vitamin Ks include phylloquinone (K1), menaquinone (K2), and menadione (K3).

As the sulfa agent, for example, sulfamethoxazole, sodium suffamethoxazole, sfluisoxazole, sodium sulfisomidin and the like can be used.

Examples of preservatives include methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, butyl paraoxybenzoate, oxyquinoline sulfate, benzalconium chloride, chlorobutanol, sodium chlorite, and benzododecinium bromide. , Sorbate, sodium dehydroacetate, benzoate, benzyl alcohol, alkylpolyaminoethylglycine, boric acid, boarite and the like can be used.

Examples of the pH adjuster include buffers such as citric acid buffer, acetate buffer, carbon dioxide buffer, borate buffer, and phosphate buffer, as well as hydrochloric acid, acetic acid, boric acid, carbonic acid, sulfuric acid, and phosphorus. Acids such as acid, citric acid and tartrate, and bases such as sodium hydroxide, sodium hydrogencarbonate, sodium carbonate, triethanolamine and monoethanolamine can be used.

Examples of the tonicity agent include sugars and salts, and examples of the salts include sodium hydrogen sulfite, sodium sulfite, potassium chloride, calcium chloride, sodium chloride, magnesium chloride, potassium acetate, sodium acetate, sodium hydrogen carbonate, and carbonic acid. Sodium, sodium thiosulfate, magnesium sulfate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate and the like can be used. As the saccharide, any monosaccharide or polysaccharide can be used, and as an example, glucose, cyclodextrin, xylitol, sorbitol, mannitol and the like can be used.

As the thickener, for example, polyvinyl alcohol, carboxyvinyl polymer, hydroxyethyl cellulose, hydroxypropylmethyl cellulose, methyl cellulose, alginic acid, hyaluronic acid, polyvinylpyrrolidone and the like or salts thereof can be used.

As the solubilizing agent, for example, nonionic surfactants such as polyoxyethylene sorbitan monooleate, polyoxyethylene hydrogenated castor oil, tyroxapol, and pluronic; polyhydric alcohols such as glycerin and macrogol can be used.

As the stabilizer, for example, polyvinylpyrrolidone, sulfite, monoethanolamine, glycerin, propylene glycol, polyethylene glycol, cyclodextrin, dextran, ascorbic acid, edetate, taurine, tocopherol and the like can be used.

Examples of the surfactant include nonionic surfactants such as tyroxalol, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene block copolymer, polyoxyethylene sorbitan fatty acid ester, and octoxinol; alkyldiaminoethylglycine, Amphoteric surfactants such as lauryldimethylaminoacetic acid betaine; anionic surfactants such as alkyl sulfates, N-acyltaurine salts, polyoxyethylene alkyl ether phosphates, polyoxyethylene alkyl ether sulfates; alkylpyridinium salts, Cationic surfactants such as alkylamine salts can be used.

Examples of fragrances or refreshing agents include menthol, ethanol, camphor, geraniol, borneol, menthol, ryuno, uikyo oil, cool mint oil, spearmint oil, peppermint oil, peppermint oil, peppermint oil, bergamot oil, eucalyptus oil and rose. Oil etc. can be used.

The optional components described above may be used for the purpose of actions other than those listed above. For example, ethanol used as a refreshing agent may be added to an ophthalmic solution as a preservative.

All references referred to herein are incorporated herein by reference in their entirety.

The embodiments of the present invention described below are for illustration purposes only and do not limit the technical scope of the present invention. The technical scope of the invention is limited only by the description of the claims. Modifications of the present invention, for example, addition, deletion and replacement of the constituent elements of the present invention may be made on condition that the gist of the present invention is not deviated.

Test Example 1: Reduction of cytotoxicity of brimonidine tartrate by blending sodium chondroitin sulfate 1. Cell culture 500 mL DMEM / F12 (Thermo Fisher Scientific), 5 mL Pen Strep (Thermo Fisher Scientific), 25 mL 5% HI FBS (Thermo Fisher Scientific), 1 mL insulin (Thermo Fisher Scientific), 1 mL insulin 1 mL of EGF (5 μg / mL) (Pepro Tech) was mixed to prepare a cell culture medium. A subcultured immortalized human corneal epithelial cell line (HCE-t strain: obtained from RIKEN BioResource Research Center) was adjusted to 5 × 10 ⁵ cells / mL in the prepared cell culture medium, and 100 μL was added to the culture well. (Corning International Inc.) was dispensed, 100 μL of DPBS (Thermo Fisher Epithelium) was added, and the cells were _{cultured in a 37 ° C. 5% CO 2} atmosphere for 16 hours or more and settled in wells.

2. 2. Cytotoxicity test (1)
Purified water is added to boric acid (Fujifilm Wako Junyaku) to dissolve it, and brimonidine tartrate (Hinewy Pharma. Tech. Co., Ltd), chondroitin sulfate sodium (malhanichiro), and sodium chloride are added, and hydrochloric acid and hydroxide are added. The pH was adjusted to 6.5 using an aqueous sodium solution to prepare a test solution having the composition shown in Table 1. The values in the table are w / v% excluding pH.

It was confirmed by a microscope that sufficient cells were present in the wells and settled in the wells, and the cell culture medium in the wells was aspirated. Each well was washed with 100 μL DPBS. 100 μL of the prepared test solution was added to the wells. Incubated in a 37 ° C. incubator for about 30 minutes. After incubation, the test solution in each well was aspirated, washed with 100 μL of DPBS, 100 μL of a solution having a ratio of DMEM / F12 90% and Cell counting kit-8 10% was added, and the mixture was incubated in an incubator at 37 ° C. for about 1 hour. After incubation, the absorbance at 450 nm was measured with a microplate reader. The test in the well containing no cells was set as low control, and the test using cell culture medium instead of the test solution was set as high control. The results are shown in FIG.

3. 3. Cell toxicity test (2) -Examination of sodium chondroitin sulfate concentration Purified water was added to boric acid (Fujifilm Wako Junyaku) to dissolve it, and brimonidine tartrate (Hinewy Pharma. Tech. Co., Ltd), chondroitin sulfate. Sodium ester (Maruhanichiro) and sodium chloride were added, and the pH was adjusted to 6.5 using hydrochloric acid and an aqueous solution of sodium hydroxide to prepare a test solution having the composition shown in Table 2. The values in the table are w / v% excluding pH.

It was confirmed by a microscope that sufficient cells were present in the well and settled in the well, and the medium in the well was aspirated. Each well was washed with 100 μL DPBS. 100 μL of the prepared test solution was added to the wells. Incubated in a 37 ° C. incubator for about 30 minutes. After incubation, the test solution in each well was aspirated, washed with 100 μL of DPBS, 100 μL of a solution having a ratio of DMEM / F12 90% and Cell counting kit-8 10% was added, and the mixture was incubated in an incubator at 37 ° C. for about 1 hour. After incubation, the absorbance at 450 nm was measured with a microplate reader. The test in the well containing no cells was set as low control, and the test using cell culture medium instead of the test solution was set as high control. The results are shown in FIG.

4. Cell toxicity test (3) -Examination of concentration of brimonidine tartrate Add purified water to boric acid (Fujifilm Wako Pure Pharmaceutical) to dissolve it, brimonidine tartrate (Hinewy Pharma. Tech. Co., Ltd), chondroitin sulfate ester. Sodium (Maruhanichiro) and sodium chloride (Fujifilm Wako Junyaku) were added, and the pH was adjusted to 6.5 using hydrochloric acid and an aqueous sodium hydroxide solution to prepare a test solution having the composition shown in Table 3. The values in the table are w / v% excluding pH.

It was confirmed by a microscope that sufficient cells were present in the well and settled in the well, and the medium in the well was aspirated. Each well was washed with 100 μL DPBS. 100 μL of the prepared test solution was added to the wells. Incubated in a 37 ° C. incubator for about 30 minutes. After incubation, the test solution in each well was aspirated, washed with 100 μL of DPBS, 100 μL of a solution having a ratio of DMEM / F12 90% and Cell counting kit-8 10% was added, and the mixture was incubated in an incubator at 37 ° C. for about 1 hour. After incubation, the absorbance at 450 nm was measured with a microplate reader. The test in the well containing no cells was set as low control, and the test using cell culture medium instead of the test solution was set as high control. The results are shown in FIG.

Test Example 2: Measurement (prescription) of the amount of brimonidine tartrate transferred to the rabbit conjunctiva Each sample was prepared by a conventional method. That is, boric acid (Fujifilm Wako Pure Chemical Industries, Ltd.), borax (Nakalaitesk), and brimonidine tartrate (Hinewy Pharma. Tech. Co., Ltd) are added and dissolved in purified water to dissolve them, and then sodium chondroitin sulfate (Maruhanichiro). Was added and dissolved, the pH was adjusted with a pH adjuster, and the mixture was mixed with purified water to prepare a test solution having the composition shown in Table 4. The values in the table are w / v% excluding pH.

(Test operation)
1-1) Tube1: Four samples in each group were dispensed into 1 mL sample tubes (1.5 mL microtubes, Zartstat).
1-2) Tube2: Four samples in each group and a sample tube with a filter (Ultrafree-MC-GV 0.45 μm, millipore) were prepared.
2) A rabbit excised eyeball (Kitayama Labes) was obtained and the conjunctiva was cut out.
3) The conjunctiva was washed by immersing it in physiological saline (Otsuka Raw Food Injection, Otsuka Pharmaceutical Factory).
4) Moisture on the surface was lightly wiped off with a Kimwipe, and the weight of the conjunctiva was measured.
5) After measuring the weight of the conjunctiva, 1-1) it was put into Tube1 and shaken at 1,500 rpm with a shaker (block bath shaker-MyBL-100CS, AS ONE) for 5 minutes.
6) The conjunctiva was taken out from the centrifuge tube of 5).
7) Take 300 μL of the sample from the centrifuge tube of 6) excluding the conjunctiva, add it to 1-2) Tube2, and centrifuge it with a desktop micro high-speed centrifuge (CT-12RE, Hitachi Koki) (15,000 rpm, 10 min). , 4 ° C), and the filtrate was used as a sample.

(Determining the amount of conjunctival transfer)
For Example 6 and Comparative Example 5, the brimonidine tartrate content of each (1) test solution without immersing the conjunctiva and (2) test operation 7) was measured by high performance liquid chromatography (HPLC) method, and (1) From the difference between 1) and (2), the amount of conjunctival transfer per sample was obtained, and from the conjunctival weight measured in 4), the content of brimonidine tartrate transferred per 1 g of conjunctiva was obtained for each individual, and the average value of 4 cases. Was taken as the amount of conjunctival transfer.
<High performance liquid chromatograph conditions>
Column: Inner diameter 4.6 mm x length 75 mm Octadecylsilylated silica gel ("SymmeteryC18 3.5" manufactured by Waters)
Detector: Ultraviolet-visible absorptiometer Measurement wavelength: 264nm
Mobile phase: 5.175 g of ammonium dihydrogen phosphate was dissolved in 900 mL of water, and 100 mL of acetonitrile for liquid chromatography was added.

［式中、Ｃ_実施例６は実施例６のブリモニジン酒石酸塩結膜移行量、Ｃ_比較例５は比較例５のブリモニジン酒石酸塩結膜移行量とした。］ The rate of increase in the amount of brimonidine tartrate transferred to the conjunctiva (C _rate ) due to the addition of sodium chondroitin sulfate was calculated according to the following formula.

[In the formula, C _{Example 6} was the brimonidine tartrate conjunctival transfer amount of Example 6, and C _{Comparative Example 5} was the brimonidine tartrate conjunctival transfer amount of Comparative Example 5. ]

The brimonidine tartrate solution was blended with sodium chondroitin sulfate to improve the transferability of brimonidine tartrate to the rabbit conjunctiva.

Claims (3)

An ophthalmic liquid preparation containing 0.01 to 0.05 w / v% of brimonidine and / or a salt thereof, and a chondroitin sulfate ester and / or a salt thereof. The liquid preparation for ophthalmology according to claim 1, wherein the salt of the chondroitin sulfate ester is a sodium salt. The liquid preparation for ophthalmology according to claim 1 or 2, wherein the content of chondroitin sulfate ester and / or a salt thereof is 0.05 to 0.5 w / v%.

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