Liquid preparations containing brimonidine

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

[Technical Field]

[0001] The present invention relates to liquid formulations containing brimonidine or a salt thereof. [Background technology]

[0002] Brimonidine and its salts are known as α-2 adrenergic receptor agonists. The human eye contains many α-2 adrenergic receptors (hereinafter sometimes abbreviated as α-2 receptors). α-2 receptor agonists reduce intraocular pressure by inhibiting aqueous humor production and promoting aqueous humor outflow via the uveoscleral outflow pathway. Based on this effect, α-2 receptor agonists have traditionally been used to treat glaucoma and ocular hypertension. Furthermore, α-2 receptor agonists reduce the lumen size of α-2 receptor-rich arterioles, particularly terminal arterioles. This action results in vasoconstriction, reducing redness and increasing whiteness of the eye, thereby improving the aesthetic appearance of the eye (Patent Document 1: Japanese Patent No. 5671459; Patent Document 2: Japanese Patent No. 5738890).

[0003] Regarding formulations containing brimonidine and / or its salts in combination with timolol and / or its salts, formulation technologies focusing on formulation stability have also been investigated. For example, Patent Document 3 (JP 2019-104727 A) discloses an aqueous liquid preparation containing brimonidine and / or its salts, a water-soluble polymer, and chlorhexidine gluconate. This document discloses that the water-soluble polymers include carboxymethylcellulose and / or its salts, polyvinyl alcohol, polyvinylpyrrolidone, hydroxypropylmethylcellulose, and hyaluronic acid, and that the inclusion of these components results in high viscosity stability and suppressed deterioration. Furthermore, Patent Document 4 (JP 2019-178083 A) discloses an aqueous ophthalmic solution containing brimonidine or its salts and a water-soluble polymer, with a pH of less than 7.4. This document discloses that examples of water-soluble polymers include polysaccharides, cellulose polymers, and synthetic polymers, and that when sodium carboxymethylcellulose, hydroxypropylmethylcellulose, and polyvinylpyrrolidone are used, the precipitation of insoluble matter is suppressed and storage stability is excellent. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 5671459 [Patent Document 2] Patent No. 5738890 [Patent Document 3] Japanese Patent Application Publication No. 2019-104727 [Patent Document 4] Japanese Patent Application Publication No. 2019-178083 Summary of the Invention [Problem to be solved by the invention]

[0005] The present invention aims to provide an ophthalmic liquid preparation having superior efficacy when brimonidine or a salt thereof is formulated into an ophthalmic liquid preparation for alleviating or suppressing eye congestion. [Means for solving the problem]

[0006] The present inventors have conducted extensive research with the aim of providing an ophthalmic liquid formulation containing low concentrations of brimonidine, and have found that the incorporation of chondroitin sulfate and / or a salt thereof reduces the cytotoxicity of the liquid formulation containing low concentrations of brimonidine. They have also found that the ophthalmic liquid formulation containing low concentrations of brimonidine to which chondroitin sulfate and / or a salt thereof has been added exhibits high conjunctival tissue penetration.

[0007] As one embodiment of the present invention, there is provided the following liquid ophthalmic formulation. [1] An ophthalmic liquid preparation containing 0.01 to 0.05 w / v% brimonidine and / or a salt thereof and chondroitin sulfate and / or a salt thereof. [2] The ophthalmic liquid formulation according to Item 1, wherein the salt of chondroitin sulfate ester is a sodium salt. [3] The ophthalmic liquid preparation according to item 1 or 2, wherein the content of chondroitin sulfate and / or a salt thereof is 0.05 to 0.5 w / v%.

[0008] Furthermore, as one embodiment of the present invention, there is provided a method for reducing cytotoxicity as described below. [4] A method for reducing the cytotoxicity of brimonidine and / or its salts when administered by eye drop administration, comprising blending chondroitin sulfate and / or its salts with an ophthalmic liquid preparation containing 0.01 to 0.05 w / v% brimonidine and / or its salts.

[0009] Furthermore, as one embodiment of the present invention, the following cytotoxicity-reducing agent is provided. [5] An agent for reducing the cytotoxicity of brimonidine and / or its salts when administered by eye drop administration, characterized by incorporating chondroitin sulfate and / or its salts into an ophthalmic liquid formulation containing 0.01 to 0.05 w / v% brimonidine and / or its salts.

[0010] Furthermore, as one embodiment of the present invention, there is provided a method for promoting conjunctival migration as described below. [6] A method for promoting the conjunctival transfer of brimonidine and / or its salts by ophthalmic administration, comprising blending chondroitin sulfate and / or its salts with an ophthalmic liquid preparation containing 0.01 to 0.05 w / v% brimonidine and / or its salts.

[0011] Furthermore, as one embodiment of the present invention, there is provided the conjunctival migration promoter shown below. [7] A conjunctival transfer promoter for brimonidine and / or its salts when administered by eye drop, characterized by incorporating chondroitin sulfate and / or its salts into an ophthalmic liquid preparation containing 0.01 to 0.05 w / v% brimonidine and / or its salts. [Effects of the Invention]

[0012] According to the present invention, an ophthalmic liquid formulation containing low concentrations of brimonidine and chondroitin sulfate and / or a salt thereof exhibits at least one of the effects of reducing the cytotoxicity of brimonidine and improving the transfer of brimonidine to conjunctival tissue. [Brief explanation of the drawings]

[0013] [Figure 1] Figure 1 is a graph showing the effects of brimonidine tartrate and chondroitin sulfate sodium on cultured cells. The vertical axis shows the absorbance of the wells after color development using the cell counting kit; higher absorbance indicates a higher number of viable cells. The low control refers to a group containing no cells, and the high control refers to a group in which cell culture medium was used instead of the drug-added test solution. While the brimonidine tartrate group (BRN alone) showed a decrease in absorbance, the chondroitin sulfate sodium group (CS alone) and the brimonidine tartrate and chondroitin sulfate sodium group (CS + BRN) showed absorbance levels similar to those of the high control. [Figure 2]Figure 2 shows the effects of brimonidine tartrate and chondroitin sulfate sodium on cultured cells. The vertical axis indicates the absorbance of the wells after color development using a cell counting kit; higher absorbance indicates a higher number of viable cells. The low control is a group without cells, and the high control is a group in which cell culture medium was used instead of the drug-containing test solution. While the brimonidine tartrate group (BRN 0.025% alone) showed a decrease in absorbance, the brimonidine tartrate and 0.1 w / v% chondroitin sulfate sodium group (CS 0.1% + BRN 0.025%), the brimonidine tartrate and 0.3 w / v% chondroitin sulfate sodium group (CS 0.3% + BRN 0.025%), and the brimonidine tartrate and 0.5% chondroitin sulfate sodium group (CS 0.5% + BRN 0.025%) showed absorbance levels comparable to those of the high control. [Figure 3] Figure 3 shows the effects of brimonidine tartrate and chondroitin sulfate sodium on cultured cells. The vertical axis indicates the absorbance of the wells after color development using a cell counting kit; higher absorbance indicates a higher number of viable cells. The low control is a group containing no cells, and the high control is a group in which cell culture medium was used instead of a drug-containing test solution. The group containing 0.5 w / v% chondroitin sulfate sodium alone (CS 0.5% alone), the group containing 0.025 w / v% brimonidine tartrate and 0.5 w / v% chondroitin sulfate sodium (CS 0.5% + BRN 0.025%), and the group containing 0.05 w / v% brimonidine tartrate and 0.5 w / v% chondroitin sulfate sodium (CS 0.5% + BRN 0.05%) showed absorbance levels comparable to those of the high control. DETAILED DESCRIPTION OF THE INVENTION

[0014] It is understood that the terms used in this specification are used in the sense commonly used in the art unless otherwise specified. Therefore, unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by those skilled in the art to which this invention belongs. The numerical range specified in this specification is intended to include its lower and upper limits.

[0015] (definition) As used herein, "brimonidine" refers to the compound with the IUPAC name 5-Bromo-N-(4,5-dihydro-1H-imidazol-2-yl)quinoxalin-6-amine. Furthermore, unless otherwise specified, the concentrations of brimonidine and / or its salts are expressed in terms of brimonidine tartrate.

[0016] As used herein, "low concentration brimonidine" refers to brimonidine at a concentration of 0.05 w / v % or less.

[0017] As used herein, the term "ophthalmic liquid preparation" refers to an aqueous liquid preparation based on water.

[0018] As used herein, the term "cytotoxicity" refers to the property of brimonidine to cause cell death, dysfunction, or growth inhibition in cells of ocular tissues.

[0019] As used herein, "reducing cytotoxicity" or "reducing cytotoxicity" refers to suppressing the effects of cytotoxicity and maintaining the inherent functions of cells.

[0020] As used herein, the term "method for reducing cytotoxicity" refers to a method for suppressing the cytotoxic effects of brimonidine on ocular tissues and for maintaining the inherent functions of cells.

[0021] As used herein, the term "cytotoxicity-reducing agent" refers to an agent that is added to suppress the cytotoxic effects of brimonidine on ocular tissues and to maintain the cells' inherent functions.

[0022] As used herein, the term "conjunctival transferability" is an index showing the ease with which brimonidine in an ophthalmic liquid preparation administered by eye drop transfers to the conjunctiva, which is the site of action.

[0023] As used herein, the term "method for promoting conjunctival transfer" refers to a method carried out to increase the amount of brimonidine in an ophthalmic liquid formulation that transfers to the conjunctiva, the site of action, when administered by eye drop.

[0024] As used herein, the term "conjunctival transport promoter" refers to an agent incorporated to increase the amount of brimonidine transported to the conjunctiva, the site of action, in an ophthalmic liquid formulation administered by eye drop.

[0025] DESCRIPTION OF THE PREFERRED EMBODIMENTS Preferred embodiments of the present invention will be described below. The embodiments provided below are provided for a better understanding of the present invention, and it is understood that the scope of the present invention should not be limited to the following description. Therefore, it is clear that those skilled in the art can make appropriate modifications within the scope of the present invention in consideration of the description in this specification. It is also understood that the following embodiments can be used alone or in combination.

[0026] In the present invention, the pharmaceutically acceptable salt of brimonidine can include any salt.The pharmaceutically acceptable salt of brimonidine can include hydrochloride, sulfate, phosphate, acetate, citrate, oxalate, malonate, salicylate, malate, fumarate, succinate, ascorbate, maleate, methanesulfonate, tartrate and other inorganic carboxylic acid salts well known to those skilled in the art, and preferably tartrate.In addition, in this specification, brimonidine and / or its salt can also be referred to as "BRN".

[0027] In the present invention, low-concentration brimonidine refers to brimonidine at a concentration of 0.05 w / v% or less. For example, the upper limit of the concentration may be 0.04 w / v% or 0.03 w / v% in order to prevent side effects. The lower limit of the concentration is not limited as long as brimonidine is contained, but may be 0.01 w / v%. For example, in consideration of the effects of the present drug, 0.015 w / v% or 0.02 w / v% may be used.

[0028] In the present invention, the chondroitin sulfate and / or its salts incorporated into the liquid preparation containing brimonidine or its salt are a type of glycosaminoglycan found in vivo. Chondroitin sulfate mainly has a structure in which the disaccharide unit D-glucuronic acid (GlcA) and N-acetyl-D-galactosamine (GalNAc) repeats, with sulfate groups at the 4th and / or 6th positions of GalNAc, but the structure may vary depending on the biological species from which it is derived. The chondroitin sulfate and / or its salts of the present invention are not limited to those in which the disaccharide unit D-glucuronic acid (GlcA) and N-acetyl-D-galactosamine (GalNAc) repeats, with sulfate groups at the 4th and / or 6th positions of GalNAc, but include all commercially available chondroitin sulfates. Chondroitin sulfate may exist in any salt form, such as sodium or potassium salts. Chondroitin sulfate and / or its salts are incorporated into eye drops to protect the corneal surface due to their moisturizing effect. In the present invention, they exhibit the effect of reducing the cytotoxicity of brimonidine or its salts. This effect can also be referred to as a cytoprotective effect. Furthermore, in the present invention, the incorporation of chondroitin sulfate or its salts enhances the conjunctival migration of brimonidine. Chondroitin sulfate and / or its salts can be incorporated 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, for example, 0.01 w / v%, 0.05 w / v%, or 0.1 w / v%. From the viewpoint of incorporation into eye drops, the upper limit of the concentration may be, for example, 3 w / v%, 1 w / v%, or 0.5 w / v%. In this specification, chondroitin sulfate and / or its salts may be abbreviated as "CS." In one embodiment, the liquid preparation of the present invention containing brimonidine or a salt thereof and chondroitin sulfate and / or a salt thereof may be characterized as not containing chlorhexidine gluconate or a salt thereof.

[0029] The conjunctiva is a membrane that covers the sclera (the white of the eye) and lines the inside of the eyelid, and is primarily composed of conjunctival epithelial cells. The conjunctival epithelial layer contains blood vessels, fibrous tissue, and lymphatic vessels. It contacts the cornea at the boundary between the white and black of the eye, and the cornea and conjunctiva constitute the outermost layer of the eye exposed to the outside world. Because the conjunctiva is exposed to the outside world, it is susceptible to bacterial and viral attack and inflammation. Even in the absence of inflammation, lack of sleep or overuse of the eyes can increase blood flow to supply oxygen and nutrients to the eyes, causing congestion. In order for brimonidine to exert its effect of reducing eye redness or whitening in the present invention, the active ingredient of the instilled formulation must reach and act on the capillaries in the conjunctiva.

[0030] Conjunctival penetration is an index showing the ease with which the active ingredient (in this invention, brimonidine or a salt thereof) of an ophthalmic liquid formulation administered by eye drops penetrates the conjunctiva, the site of action. Corneal epithelial cells and conjunctival epithelial cells form tight junctions between the cells, forming hydrophobic membranes originating from the cell membrane. This restricts the permeation of water-soluble drugs through the cornea and conjunctiva. The intraocular penetration of an instilled drug via the cornea or conjunctiva is determined by measuring the drug concentration in the aqueous humor or conjunctival tissue after instillation. However, since measurement in humans is difficult, analysis using a predictive model based on animal experiments is performed. A simple method for measuring intraocular penetration is known, based on the octanol / water partition coefficient. The octanol / water partition coefficient can be used to evaluate not only intraocular penetration but also conjunctival penetration.

[0031] In the present invention, the ophthalmic liquid preparation is an aqueous liquid preparation mainly based on water, but may further contain any liquid base that can be used in eye drops. The ophthalmic liquid preparation of the present invention is prepared so as to have a pH and osmolality acceptable for eye drops. The pH of the ophthalmic liquid preparation can be adjusted to 5.0 to 9.0, for example, 5.5 to 8.5, using a pH adjuster. It can also be adjusted to 6.0 to 8.0. The amounts of components added to the ophthalmic liquid preparation are adjusted so that the osmolality ratio of the ophthalmic liquid preparation is preferably 0.5 to 2.5, more preferably 0.7 to 1.5, for example.

[0032] The ophthalmic liquid preparation of the present invention is preferably an eye drop. The ophthalmic liquid preparation of the present invention may be a liquid preparation that relieves or suppresses eye redness. Relieving or suppressing eye redness refers to increasing the whiteness of the white of the eye, which can also be called eye whitening.

[0033] The ophthalmic liquid formulation of the present invention may contain any component that can be used in eye drops, provided that the effects of the present invention are not impaired. In addition to brimonidine tartrate, the active ingredient of the present invention, optional active ingredients and additives may be included. Examples of such ingredients include, but are not limited to, decongestants, focus-adjusting agents, anti-inflammatory and astringent agents, antihistamines, vitamins, nutrients, sulfonamides, preservatives, pH adjusters, isotonicity agents, thickeners, antioxidants, solubilizers, stabilizers, surfactants, fragrances, and refreshing agents. These active ingredients and additives may be used singly or in combination from each category.

[0034] As the decongestant, for example, epinephrine, ephedrine, tetrahydrozoline, naphazoline, phenylephrine, methylephedrine, or salts thereof may be used.

[0035] Neostigmine methylsulfate can be used as the accommodative agent.

[0036] Examples of anti-inflammatory and astringent agents that can be used include ε-aminocaproic acid, allantoin, berberine or a salt thereof, azulene sulfonic acid or a salt thereof, glycyrrhizic acid or a salt thereof, zinc sulfate, zinc lactate, and lysozyme chloride.

[0037] As the antihistamine, diphenhydramine hydrochloride and chlorpheniramine maleate can be used.

[0038] Amino acids or salts thereof can be used as nutritional components. The amino acids or salts thereof include, in addition to amino acids, substances having a sulfate group instead of the carboxyl group of an amino acid, such as taurine. Examples of amino acids include glycine, alanine, methionine, valine, threonine, glutamine, glutamic acid, asparagine, aspartic acid, cysteine, histidine, isoleucine, leucine, lysine, phenylalanine, tryptophan, arginine, proline, tyrosine, and serine. Preferred amino acids include aspartic acid, methionine, and glycine. The amino acids, excluding glycine, may be L-amino acids, D-amino acids, or DL-amino acids.

[0039] Vitamins are a general term for organic compounds other than carbohydrates, proteins, and lipids that are essential nutrients in trace amounts for the survival and growth of living organisms but cannot be synthesized in sufficient quantities within the organism's body. Vitamins are broadly divided into water-soluble vitamins and fat-soluble vitamins. Water-soluble vitamins include B vitamins and vitamin C (ascorbic acid). Fat-soluble vitamins include vitamins A, D, E, and K. The over-the-counter drug manufacturing (import) approval standards stipulate the vitamins that may be incorporated into eye drops. From this perspective, vitamins A, B, and E are particularly preferred.

[0040] Examples of vitamin A include retinol and its related substances. Examples of retinol-related substances include retinal, retinoic acid, retinol palmitate, and other retinoids, such as isotretinoin, alitretinoin, acitretin, etretinate, adapalene, tazarotene, and bexarotene. From the perspective of formulation into eye drops, retinol palmitate and retinol acetate are preferred. Since vitamin A acts on epithelial cells to induce their proliferation, it can be incorporated into eye drops for the purpose of protecting the cornea and conjunctiva. It can also be incorporated into eye drops for the treatment of ophthalmic diseases such as night blindness, xeroconjunctival disease, xerocorneal disease, and keratomalacia.

[0041] Examples of B vitamins that can be used include vitamin B1 (thiamine, etc.), vitamin B2, vitamin B3 (niacin, etc.), vitamin B5, vitamin B6, vitamin B7 (biotin, etc.), vitamin B9 (folic acid, etc.), and vitamin B12. Vitamins include provitamins, which are derivatives, and pharmaceutically acceptable salts. Among the B vitamins, vitamins B2, B5, B6, and B12 are preferred, particularly from the viewpoint of being incorporated into eye drops.

[0042] Vitamin B2 may be riboflavin, riboflavin phosphate, riboflavin butyrate, riboflavin acetate, flavin adenine dinucleotide, flavin mononucleotide, or pharmaceutically acceptable salts thereof. Examples of salts include sodium and potassium salts. Flavin adenine dinucleotide sodium is preferred for use in eye drops. Vitamin B2 is directly involved in oxidation-reduction reactions, and when used in eye drops, it promotes the enzymatic respiratory metabolism of corneal and conjunctival cells, thereby protecting the cornea and conjunctiva. Vitamin B2 can also be incorporated into eye drops to treat keratitis thought to be caused by vitamin B2 deficiency or metabolic disorders.

[0043] As vitamin B5, panthenol, pantothenic acid, or derivatives or salts thereof can be used. In addition to panthenol and pantothenic acid, examples of derivatives or salts thereof include pantethine, pantetheine, pantothenyl alcohol, pantothenyl ethyl ether, pantetheine pantothenyl alcohol, calcium pantothenate, and sodium pantothenate. From the viewpoint of use as an eye drop, panthenol, calcium pantothenate, and sodium pantothenate are preferred as vitamin B5.

[0044] Vitamin B6 may be pyridoxal, pyridoxamine, pyridoxine, or a pharmaceutically acceptable salt thereof. From the viewpoint of use as an eye drop, pyridoxine hydrochloride is preferred. Vitamin B6 is involved in protein metabolism as a coenzyme for amino acid decarboxylase and aminotransferase in the body, and can be incorporated into eye drops to suppress eye fatigue.

[0045] Vitamin B12 is a compound with a structure in which cobalt is coordinated to a corrin ring, and specific examples include cyanocobalamin, mecobalamin (methylcobalamin), hydroxocobalamin, adenosylcobalamin, hydroxocobalamin hydrochloride, hydroxocobalamin acetate, etc. Vitamin B12 can be incorporated into eye drops for its pharmacological effects, such as improving tired eyes and eye strain.

[0046] Examples of vitamin E include tocopherol, tocotrienol, tocophersolan, and derivatives thereof. Tocopherol and tocotrienol may be α-, β-, γ-, or δ-tocopherol, and may be either d- or dl-tocopherol. For use as an eye drop, d-α-tocopherol acetate is an example.

[0047] Vitamin C includes ascorbic acid and its salts. Vitamin D compounds include vitamin D2 (ergosterol, ergocalciferol), D3 (7-dehydrocholesterol), previtamin D3 (cholecalciferol, 25-hydroxycholecalciferol, calcitriol (1,25-dihydroxycholecalciferol), calcitronic acid), vitamin D4 (dihydroergocalciferol), and vitamin D5 (dihydrotachysterol, calcipotriol, tacalcitol, paricalcitol). Vitamin K compounds include phylloquinone (K1), menaquinone (K2), and menadione (K3).

[0048] Examples of sulfonamide drugs that can be used include sulfamethoxazole, sulfamethoxazole sodium, sulfisoxazole, and sulfisomidine sodium.

[0049] Examples of preservatives that can be used include methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, butyl parahydroxybenzoate, oxyquinoline sulfate, benzalkonium chloride, chlorobutanol, sodium chlorite, benzododecinium bromide, sorbate, sodium dehydroacetate, benzoate, benzyl alcohol, alkylpolyaminoethylglycine, boric acid, and borax.

[0050] Examples of pH adjusters that can be used include buffers such as citrate buffers, acetate buffers, carbonate buffers, borate buffers, and phosphate buffers, as well as acids such as hydrochloric acid, acetic acid, boric acid, carbonic acid, sulfuric acid, phosphoric acid, citric acid, and tartaric acid, and bases such as sodium hydroxide, sodium bicarbonate, sodium carbonate, triethanolamine, and monoethanolamine.

[0051] Examples of the isotonic agent include sugars and salts, and examples of the salts that can be used include sodium hydrogen sulfite, sodium sulfite, potassium chloride, calcium chloride, sodium chloride, magnesium chloride, potassium acetate, sodium acetate, sodium hydrogen carbonate, sodium carbonate, sodium thiosulfate, magnesium sulfate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, etc. As the sugar, any monosaccharide or polysaccharide can be used, and examples of the sugar that can be used include glucose, cyclodextrin, xylitol, sorbitol, mannitol, etc.

[0052] Examples of thickeners that can be used include polyvinyl alcohol, carboxyvinyl polymer, hydroxyethyl cellulose, hydroxypropylmethyl cellulose, methyl cellulose, alginic acid, hyaluronic acid, polyvinylpyrrolidone, and salts thereof.

[0053] Examples of solubilizing agents that can be used include nonionic surfactants such as polyoxyethylene sorbitan monooleate, polyoxyethylene hydrogenated castor oil, tyloxapol, and Pluronic; and polyhydric alcohols such as glycerin and macrogol.

[0054] Examples of stabilizers that can be used include polyvinylpyrrolidone, sulfites, monoethanolamine, glycerin, propylene glycol, polyethylene glycol, cyclodextrin, dextran, ascorbic acid, edetate, taurine, and tocopherol.

[0055] Examples of surfactants that can be used include nonionic surfactants such as tyloxapol, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene block copolymers, polyoxyethylene sorbitan fatty acid esters, and octoxynol; amphoteric surfactants such as alkyldiaminoethylglycine and lauryldimethylaminoacetic acid betaine; anionic surfactants such as alkyl sulfates, N-acyltaurine salts, polyoxyethylene alkyl ether phosphates, and polyoxyethylene alkyl ether sulfates; and cationic surfactants such as alkylpyridinium salts and alkylamine salts.

[0056] Examples of fragrances or freshening agents that can be used include menthol, ethanol, camphor, geraniol, borneol, menthol, rhubarb, fennel oil, cool mint oil, spearmint oil, peppermint water, peppermint oil, bergamot oil, eucalyptus oil, and rose oil.

[0057] The optional ingredients mentioned above may be used for purposes other than those listed above. For example, ethanol, which is used as a cooling agent, may be added to ophthalmic solutions as a preservative.

[0058] All documents mentioned herein are incorporated by reference in their entirety.

[0059] The following examples of the present invention are for illustrative purposes only and do not limit the technical scope of the present invention. The technical scope of the present invention is limited only by the claims. The present invention may be modified, for example, by adding, deleting, or substituting components of the present invention, provided that the modifications do not depart from the spirit of the present invention. [Example]

[0060] Test Example 1: Reduction of cytotoxicity of brimonidine tartrate by adding sodium chondroitin sulfate 1.Cell culture Cell culture medium was prepared by mixing 500 mL of DMEM / F12 (Thermo Fisher Scientific), 5 mL of Pen Strep (Thermo Fisher Scientific), 25 mL of 5% HI FBS (Thermo Fisher Scientific), 1 mL of insulin (5 μg / mL) (Fujifilm Wako), and 1 mL of EGF (5 μg / mL) (Pepro Tech). A subcultured immortalized human corneal epithelial cell line (HCE-t line: obtained from RIKEN BioResource Research Center) was placed at 5 × 10 in the prepared cell culture medium. 5 The concentration was adjusted to cells / mL, and 100 μL was dispensed into a culture well (Corning International Inc.). 100 μL of DPBS (Thermo Fisher Scientific) was added, and the cells were cultured at 37°C in a 5% CO atmosphere for 16 hours or more to allow the cells to settle in the well.

[0061] 2. Cytotoxicity test (1) Purified water was added to boric acid (Fujifilm Wako Pure Chemical Industries, Ltd.) to dissolve it, and brimonidine tartrate (Hinewy Pharma.Tech.Co.,Ltd.), sodium chondroitin sulfate (Maruha Nichiro), and sodium chloride were added. The pH was adjusted to 6.5 using hydrochloric acid and aqueous sodium hydroxide to prepare a test solution with the composition shown in Table 1. The values ​​in the table, except for pH, are in w / v%. [Table 1]

[0062] After confirming the presence and adhesion of sufficient cells in the wells under a microscope, the cell culture medium was aspirated. Each well was washed with 100 μL of DPBS. 100 μL of the prepared test solution was added to the well. The wells were incubated in a 37°C incubator for approximately 30 minutes. After incubation, the test solution was aspirated from each well, washed with 100 μL of DPBS, and 100 μL of a solution consisting of 90% DMEM / F12 and 10% Cell Counting Kit-8 was added. The wells were incubated in a 37°C incubator for approximately 1 hour. After incubation, the absorbance at 450 nm was measured using a microplate reader. A test using a well without cells served as the low control, and a test using cell culture medium instead of the test solution served as the high control. The results are shown in Figure 1.

[0063] 3. Cytotoxicity Test (2) - Examination of the Concentration of Sodium Chondroitin Sulfate Boric acid (Fujifilm Wako Pure Chemical Industries) was dissolved in purified water, and brimonidine tartrate (Hinewy Pharma.Tech.Co.,Ltd), chondroitin sulfate sodium (Maruha Nichiro), and sodium chloride were added, and the pH was adjusted to 6.5 using hydrochloric acid and aqueous sodium hydroxide to prepare a test solution with the composition shown in Table 2. The values ​​in the table are in w / v% except for pH. [Table 2]

[0064] After confirming the presence and adhesion of sufficient cells in the wells under a microscope, the medium in the wells was aspirated. Each well was washed with 100 μL of DPBS. 100 μL of the prepared test solution was added to the wells. The wells were incubated in a 37°C incubator for approximately 30 minutes. After incubation, the test solution in each well was aspirated, washed with 100 μL of DPBS, and 100 μL of a solution consisting of 90% DMEM / F12 and 10% Cell Counting Kit-8 was added. The wells were incubated in a 37°C incubator for approximately 1 hour. After incubation, the absorbance at 450 nm was measured using a microplate reader. A test using a well without cells served as the low control, and a test using cell culture medium instead of the test solution served as the high control. The results are shown in Figure 2.

[0065] 4. Cytotoxicity Test (3) - Examination of Brimonidine Tartrate Concentration Purified water was added to boric acid (Fujifilm Wako Pure Chemical Industries) to dissolve it, and brimonidine tartrate (Hinewy Pharma.Tech.Co.,Ltd), chondroitin sulfate sodium (Maruha Nichiro), and sodium chloride (Fujifilm Wako Pure Chemical Industries) were added. The pH was adjusted to 6.5 using hydrochloric acid and aqueous sodium hydroxide to prepare a test solution with the composition shown in Table 3. The values ​​in the table are in w / v% except for pH. [Table 3]

[0066] After confirming the presence and adhesion of sufficient cells in the wells under a microscope, the medium in the wells was aspirated. Each well was washed with 100 μL of DPBS. 100 μL of the prepared test solution was added to the wells. The wells were incubated in a 37°C incubator for approximately 30 minutes. After incubation, the test solution in each well was aspirated, washed with 100 μL of DPBS, and 100 μL of a solution consisting of 90% DMEM / F12 and 10% Cell Counting Kit-8 was added. The wells were incubated in a 37°C incubator for approximately 1 hour. After incubation, the absorbance at 450 nm was measured using a microplate reader. A test using a well without cells served as the low control, and a test using cell culture medium instead of the test solution served as the high control. The results are shown in Figure 3.

[0067] Test Example 2: Measurement of the amount of brimonidine tartrate transferred to rabbit conjunctiva (Prescription) Each sample was prepared by the usual method. Specifically, boric acid (Fujifilm Wako Pure Chemical Industries, Ltd.), borax (Nacalai Tesque), and brimonidine tartrate (Hinewy Pharma. Tech. Co., Ltd.) were added to purified water and dissolved, and then sodium chondroitin sulfate (Maruha Nichiro) was added and dissolved. The pH was then adjusted with a pH adjuster, and the solution was diluted with purified water to prepare the test solution with the composition shown in Table 4. The values ​​in the table are in w / v% except for pH. [Table 4]

[0068] (Test operation) 1-1) Tube 1: Four tubes for each group. The specimen was dispensed into 1 mL sample tubes (1.5 mL microtubes, Zaltostat). 1-2) Tube 2: Four sample tubes with filters (Ultrafree-MC-GV 0.45 μm, Millipore) were prepared for each group. 2) Rabbit eyeballs (Kitayama Labes) were obtained and the conjunctiva was excised. 3) The conjunctiva was washed by immersion in physiological saline (Otsuka saline injection, Otsuka Pharmaceutical Factory). 4) The moisture on the surface was gently wiped off with a Kimwipe, and the conjunctival weight was measured. 5) After measuring the conjunctival weight, the conjunctiva was placed in 1-1) Tube 1 and shaken at 1,500 rpm for 5 minutes using a shaker (Block Bath Shaker - MyBL-100CS, AS ONE). 6) The conjunctiva was removed from the centrifuge tube in 5). 7) 300 μL of the sample was taken from the centrifuge tube in 6) after removing the conjunctiva, added to 1-2) Tube 2, and centrifuged (15,000 rpm, 10 min, 4°C) in a tabletop high-speed microcentrifuge (CT-12RE, Hitachi Koki), and the filtrate was used as the sample.

[0069] (Determination of the amount of transfer to the conjunctiva) For Example 6 and Comparative Example 5, the brimonidine tartrate content of each of (1) the test solution without immersion in the conjunctiva and (2) the specimens of test procedure 7) was measured by high-performance liquid chromatography (HPLC), and the conjunctival transfer amount per specimen was calculated from the difference between (1) and (2). Furthermore, the conjunctival weight measured in 4) was used to calculate the brimonidine tartrate content per 1 g of conjunctiva for each individual, and the average value of the four specimens was used as the conjunctival transfer amount. <High-performance liquid chromatography conditions> Column: 4.6 mm inner diameter x 75 mm length, octadecylsilanized silica gel ("Symmetry C18 3.5" manufactured by Waters) Detector: UV-visible spectrophotometer 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.

[0070] Increase in the amount of brimonidine tartrate transferred to the conjunctiva by the addition of sodium chondroitin sulfate (C rate ) was calculated according to the following formula:

number

[0071] [Table 5]

[0072] The addition of sodium chondroitin sulfate to the brimonidine tartrate solution improved the transfer of brimonidine tartrate to the rabbit conjunctiva.

Claims

1. An ophthalmic liquid preparation containing 0.01 to 0.05 w / v % of brimonidine and / or a salt thereof and chondroitin sulfate and / or a salt thereof.

2. 2. The ophthalmic liquid formulation according to claim 1, wherein the salt of chondroitin sulfate ester is a sodium salt.

3. 3. The liquid ophthalmic preparation according to claim 1, wherein the content of chondroitin sulfate and / or a salt thereof is 0.05 to 0.5 w / v %.

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