TWI743858B - Methods to stabilize loxidine in water-based eye drops - Google Patents

Abstract

The present invention relates to diquafosol (diquafosol) or its salt containing 0.1-10% (w/v) concentration, and chlorhexidine (chlorhexidine) with a concentration of 0.0001-10% (w/v) Water-based eye drops.

Description

Methods to stabilize loxidine in water-based eye drops

The present invention relates to an aqueous eyedrop, which is an aqueous eyedrop containing diquafosol or its salt at a concentration of 0.1-10% (w/v) (hereinafter, also referred to as diquafosol) Eye drops), and are aqueous eye drops containing chlorhexidine at a concentration of 0.0001 to 0.1% (w/v).

Generally speaking, there are types of eye drops that are used multiple times within a certain period of time after opening (multi-dose eye drops) and one-time use (single-dose eye drops). In particular, in order to prevent product corruption due to microbial contamination during use, and to ensure the storage stability of eye drops, preservatives are usually included in multi-dose eye drops.

As for preservatives, benzalkonium chloride (hereinafter, also referred to as BAK), which has an excellent antiseptic effect, has been widely used. On the other hand, it is known that when BAK is used in a high concentration, it may cause corneal disorders. In addition, it is also accused that when an eye drop containing BAK is dripped into the user's eye wearing a soft contact lens, BAK will contact the contact lens and deform the soft contact lens itself. Therefore, it is generally prohibited to instill eye drops containing BAK while wearing soft contact lenses.

In addition, diquafoxol is also known as ^{purinergic receptor agonists such as P 1} , P ⁴ -bis(uridine-5') tetraphosphate, Up ₄ U, etc. In Japan, it contains 3% (w/v) The concentration of diquafoxol sodium eye drops (product name: Diquas ^{( registered trademark )} eye drops 3%) has been used in the treatment of dry eye. Here, Diquas ^{( registered trademark )} drops 3% eye drops and contains BAK for the above reasons.

In addition, Patent Document 1 has disclosed an aqueous eye drop containing diquafosol or its salt at a concentration of 0.1-10% (w/v) and a chelating agent at a concentration of 0.0001-1% (w/v). [Prior Technical Literature] [Patent Literature]

[Patent Document 1] JP 2013-227291 A

[The problem to be solved by the invention]

The subject of the present invention is to provide a diquafox ophthalmic solution that does not contain BAK and has higher safety. Furthermore, the subject of the present invention is to provide quafosol eye drops containing a preservative that is physically and chemically stable during preparation and long-term storage of eye drops. [Means to solve the problem]

The inventors of the present invention conducted intensive studies to solve the above-mentioned problems and found that diquafosol or its salt at a concentration of 0.1 to 10% (w/v) and a concentration of 0.0001 to 0.1% (w/v) Lohexidine-based aqueous eye drops (hereinafter, also referred to as "this eye drops") have excellent preservation efficiency, and the present invention has been completed. Furthermore, the present inventors found that this ophthalmic solution suppresses the deformation of soft contact lenses. In addition, the present inventors found that the present eyedrops are better for culturing immortalized human corneal epithelial cells than eyedrops containing BAK and eyedrops that do not contain diquafosol or its salt and contain loxidine. Shows high viable cell activity. In addition, the inventors found that the present eye drops significantly increase NIBUT (non-invasive liquid layer break time) in eyes wearing soft contact lenses, that is, in eyes wearing soft contact lenses, The liquid layer is stabilized. On the other hand, such an effect has not been observed with artificial liquid. Since the occurrence/exacerbation of dry eye symptoms with soft contact lenses is due to the decrease in the stability of the liquid layer, the stabilization of the liquid layer caused by this ophthalmic solution can be used in the eyes with soft contact lenses Prevention and/or treatment of dry eye. In addition, the eye drops are also useful for the prevention and/or treatment of eye dryness and/or eye discomfort in eyes wearing soft contact lenses. Furthermore, the present inventors have found that loxidine-like quafosine ophthalmic solution containing a specific concentration range of 0.001 to 0.005% (w/v) maintains high levels of ophthalmic solution during preparation. The survival rate of hexidine, and the lohexidine in this eyedrop has excellent long-term stability.

That is, the present invention provides the aqueous eye drops disclosed below. (1) An aqueous eyedrop containing diquafosol or its salt at a concentration of 0.1-10% (w/v) and loxidine at a concentration of 0.0001-0.1% (w/v). (2) The aqueous eye drops as described in (1), wherein the loxidine is chlorhexidine gluconate. (3) The aqueous eye drops as described in (1) or (2), wherein the concentration of loxidine in the eye drops is 0.0005 to 0.05% (w/v). (4) The aqueous eye drops as described in (1) or (2), wherein the concentration of loxidine in the eye drops is 0.001 to 0.005% (w/v). (5) The aqueous eye drops as described in (1) or (2), wherein the concentration of loxidine in the eye drops is greater than 0.001% (w/v) to less than 0.005% (w/v) . (6) The aqueous eye drops as described in (1) or (2), wherein the concentration of loxidine in the eye drops is 0.0015 to 0.005% (w/v). (7) The aqueous eye drops as described in (1) or (2), wherein the concentration of loxidine in the eye drops is 0.002 to 0.005% (w/v). (8) The aqueous eyedrops as described in any one of (1) to (7), wherein the concentration of diquafosol or its salt in the eyedrops is 3% (w/v). (9) The aqueous eye drops as described in any one of (1) to (8), which further contains a chelating agent. (10) The aqueous eye drops as described in (9), wherein the chelating agent is edetic acid or a salt thereof. (11) The aqueous eye drops as described in any one of (1) to (10), which is for soft contact lenses. (12) The aqueous eye drops as described in (11), wherein the soft contact lens is a silicone hydrogel contact lens.

In addition, the present invention provides methods for suppressing deformation of soft contact lenses disclosed below. (13) A method for suppressing the deformation of soft contact lenses, which uses the aqueous eye drops as described in any one of (1) to (12).

Furthermore, the present invention also relates to the following. (14) An aqueous ophthalmic solution for the prevention and/or treatment of dry eye, which contains diquafosol or its salt at a concentration of 0.1-10% (w/v), and 0.0001-0.1% (w/v) v) Lohexidine class of concentration. (15) Use of an aqueous eyedrop for the prevention and/or treatment of dry eye, which contains diquafosol or its salt at a concentration of 0.1-10% (w/v), and 0.0001-0.1% ( w/v) Lohexidine class of concentration. (16) A method for preventing and/or treating dry eye, which comprises administering diquafosol or its salt at a concentration of 0.1-10% (w/v), and 0.0001-0.1% (w/v) The concentration of lohxidine-like water-based eye drops.

In addition, the present invention provides the aqueous eye drops disclosed below. (17) The aqueous eyedrops according to any one of (1) to (12), which are used in the eyes with soft contact lenses for the prevention and/or treatment of dry eye. (18) The aqueous eye drops as described in any one of (1) to (12), which are used in the eyes with soft contact lenses to improve the stability of the liquid layer. (19) The aqueous eyedrops according to any one of (1) to (12), which are used in the eyes with soft contact lenses for the prevention or treatment of eye dryness or eye discomfort. (20) The aqueous eye drops according to any one of (17) to (19), wherein the soft contact lens is a silicone hydrogel contact lens.

In addition, the present invention provides the method disclosed below. (21) A method for suppressing the decrease in the residual rate of lohexidine in aqueous eye drops, which is used in diquafosol or its salt and lohexidine at a concentration of 0.1 to 10% (w/v) The preparation of a fixed type of water-based eye drops uses loxidine at a concentration of 0.0001 to 0.005% (w/v) to suppress the decrease in the residual rate of loxidine in the water-based eye drops . (22) A method of stabilizing loxidine in an aqueous ophthalmic solution containing diquafosol or its salt at a concentration of 0.1-10% (w/v) and loxidine According to the method, the concentration of the loxidine is greater than 0.001% (w/v) to less than 0.1% (w/v). [Effects of Invention]

It is clear from the results of the test examples described later that this ophthalmic solution has an excellent preservation effect. Furthermore, this eye drops can be used for soft contact lenses because it inhibits the deformation of soft contact lenses. In addition, compared with the eyedrops containing BAK and the eyedrops containing no diquafosol or its salt and containing lohexidine, this eyedrop showed higher viable cells in cultured immortalized human corneal epithelial cells. active. According to this, this ophthalmic solution is highly safe for living organisms, especially for corneal and conjunctival epithelium, and is useful for use in diseases such as dry eye and other unstable corneal and conjunctival epithelium. In addition, the eye drops are attached to the eyes wearing soft contact lenses, which significantly raises NIBUT. On the other hand, no such effect has been observed with artificial liquid. In other words, the eye drops are attached to the eyes wearing soft contact lenses to stabilize the liquid layer. The occurrence/exacerbation of dry eye symptoms caused by wearing soft contact lenses is due to the decrease in the stability of the liquid layer. The stabilization of the liquid layer caused by the eye drops is caused by wearing soft contact lenses. It is useful for the prevention and/or treatment of dry eye in the eyes. In addition, the eye drops are also useful for the prevention and/or treatment of eye dryness and/or eye discomfort in eyes wearing soft contact lenses. Furthermore, the diquafosine ophthalmic solution containing loxidine in a specific concentration range, that is, 0.001-0.005% (w/v) concentration, maintains galloxidine during the preparation of the ophthalmic solution. The residual rate of the serotonin, and the loxidine in the eye drops has excellent long-term stability.

[Form to implement the invention]

Those related to the present invention will be described in further detail.

Diquafox is a compound represented by the following chemical structural formula.

Regarding the "salt of diquafosol", there is no particular limitation if it is a medically acceptable salt, and examples include metal salts with lithium, sodium, potassium, calcium, magnesium, zinc, etc.; with hydrochloric acid, hydrobromic acid, Salts of inorganic acids such as hydroiodic acid, nitric acid, sulfuric acid, phosphoric acid; and acetic acid, fumaric acid, maleic acid, succinic acid, citric acid, tartaric acid, adipic acid, gluconic acid, grape heptanoic acid, Glucuronic acid, terephthalic acid, methanesulfonic acid, lactic acid, hippuric acid, 1,2-ethanedisulfonic acid, isethionic acid, lactobionic acid, oleic acid, hydroxynaphthoic acid, polygalacturonic acid, hard Fatty acid, tannic acid, trifluoromethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, lauryl sulfate, methyl sulfate, naphthalenesulfonic acid, sulfosalic acid and other organic acid salts; and methyl bromide, methyl iodide, etc. Quaternary ammonium salt; salt with bromide, chloride, iodide, etc.; salt with ammonia; with triethylenediamine, 2-aminoethanol, 2,2-iminobis(ethanol) , 1-Deoxy-1-(methylamino)-2-D-sorbitol, 2-amino-2-(hydroxymethyl)-1,3-propanediol, procaine, N, Salts of organic amines such as N-bis(phenylmethyl)-1,2-ethaneamine diamine.

In the present invention, "diquafosol or its salt" also includes the hydrate and organic solvate of diquafosol (free form) or its salt.

In the case where diquafoxol or its salt has polymorphs and polymorph groups (polymorph systems), these polymorphs and polymorph groups (polymorph systems) are also included in the present invention The scope. In this article, the polymorphic group (polymorphic system) refers to the various crystal forms and their respective crystal forms in each stage of the situation where the crystal form has changed according to the conditions and states of the production, crystallization, and storage of the crystals. The whole process.

As a preferred sodium salt of quarafoxol of the "diquafoxol or its salt" of the present invention, the tetrasodium salt of quarafoxol represented by the following chemical structural formula (hereinafter, also simply referred to as "diquafoxol" Sodium ") is particularly good.

Diquafoxol or its salt can be produced by the method disclosed in Japanese Special Publication No. 2001-510484.

The eye drops may also contain active ingredients other than diquafosol or its salt, but it is preferable to contain diquafosol or its salt as the only active ingredient.

The concentration of quafoxol or its salt in the eye drops is 0.1-10%(w/v), but 1-10%(w/v) is better, 3%(w/v) is particularly good .

In the present invention, "aqueous eye drops" means eye drops that use water as a solvent (base).

In the present invention, lohexidine and its salts are included in the "lohexidine class". Loxidine is a compound represented by the following chemical structural formula, and is also called 1,1'-hexamethylene bis[5-(4-chlorophenyl)biguanide] compound.

啉、哌

、吡咯啶、三吡啶、甲吡啶等之有機胺之鹽等)、與無機鹼之鹽[例如，銨鹽；與鹼金屬(鈉、鉀等)、鹼土類金屬(鈣、鎂等)、鋁等之金屬之鹽等]等。此等之鹽尤列舉較佳為有機酸鹽及/或無機酸鹽，更較佳為羥基羧酸鹽、單羧酸鹽及/或無機酸鹽，進一步較佳為葡糖酸鹽、乙酸鹽及/或鹽酸鹽，特佳為葡糖酸鹽。此等之洛赫西定之鹽可以1種單獨使用，亦可將2種以上任意組合來使用。In the present invention, within the above-mentioned lohexidine, the "salt of lohexidine" is not particularly limited as long as it is a pharmaceutically acceptable salt. Specifically, organic acid salts [e.g., single Carboxylate (acetate, trifluoroacetate, butyrate, palmitate, stearate, etc.), polycarboxylate (fumarate, maleate, succinate, Malonate, etc.), hydroxycarboxylate (gluconate, lactate, tartrate, citrate, etc.), organic sulfonate (methanesulfonate, toluenesulfonate, toluenesulfonate) Salt (tosylate, etc.), etc.], inorganic acid salts (for example, hydrochloride, sulfate, nitrate, hydrobromide, phosphate, etc.), salts with organic bases (for example, with methylamine, triethyl Base amine, triethanolamine,

Morpholine, piperazine

, Pyrrolidine, tripyridine, picoline and other organic amine salts, etc.), salts with inorganic bases [e.g., ammonium salts; with alkali metals (sodium, potassium, etc.), alkaline earth metals (calcium, magnesium, etc.), aluminum And other metal salts, etc.] and so on. In particular, these salts are preferably organic acid salts and/or inorganic acid salts, more preferably hydroxycarboxylates, monocarboxylates and/or inorganic acid salts, and still more preferably gluconates and acetates. And/or hydrochloride, particularly preferably gluconate. These loxidine salts can be used alone or in any combination of two or more.

Loxidine and its salts can be synthesized by well-known methods, and can also be obtained as commercially available products.

The concentration of loxidine in this eyedrop is 0.0001～0.1%, but 0.0005～0.05%(w/v) is better, 0.001～0.005%(w/v) or greater than 0.001%(w/v) ) To less than 0.005% (w/v), preferably 0.0015 to 0.005% (w/v), more preferably 0.002 to 0.005% (w/v). More specifically, 0.0015(w/v)%, 0.0020(w/v)%, 0.0025(w/v)%, 0.0030(w/v)%, 0.0035(w/v)%, 0.0040(w/v) )%, 0.0045(w/v)%, 0.0050(w/v)% are preferred.

In the present invention, the "chelating agent" is not particularly limited as long as it is a compound that chelates metal ions, but for example, ethylene diamine tetraacetic acid (ethylene diamine tetraacetic acid), ethylene tetraacetic acid Amine monosodium, disodium ethylenediamine tetraacetate, trisodium ethylenediamine tetraacetate, tetrasodium ethylenediamine tetraacetate, potassium ethylenediamine tetraacetate, tripotassium ethylenediamine tetraacetate, ethylenediamine tetraacetate Potassium and other ethylene diamine tetraacetic acid or its salts; citric acid, monosodium citrate, disodium citrate, trisodium citrate, monopotassium citrate, dipotassium citrate, tripotassium citrate and other citric acid or its salts Salt; Metaphosphoric acid or its salt such as metaphosphoric acid, sodium metaphosphate, potassium metaphosphate; Pyrophosphoric acid or its salt such as pyrophosphate, tetrasodium pyrophosphate, tetrapotassium pyrophosphate, etc.; Polyphosphoric acid, sodium polyphosphate, potassium polyphosphate Polyphosphoric acid or its salt; monosodium malate, disodium malate, monopotassium malate, dipotassium malate, etc. malic acid or its salt; sodium tartrate, potassium tartrate, potassium sodium tartrate, etc. tartaric acid or its salt ; Phytic acid (phytic acid) sodium, potassium phytate and other phytic acid or its salt. In addition, in the present invention, "ethylenediamine tetraacetate, citric acid, metaphosphoric acid, pyrophosphoric acid, polyphosphoric acid, malic acid, tartaric acid, phytic acid and their salts" also include their respective free bodies or their salts The hydrate and organic solvate.

In the present invention, the chelating agent is ethylene diamine tetraacetic acid, ethylene diamine tetraacetic acid salt (ethylene diamine tetraacetic acid salt), citric acid, citric acid salt (citrate), metaphosphoric acid, metaphosphoric acid salt ( Metaphosphate), polyphosphoric acid, and salt of polyphosphoric acid (polyphosphate) are preferred, sodium salt of ethylene diamine tetraacetate (including hydrate of disodium ethylene diamine tetraacetate), citric acid (including Hydrates such as citric acid monohydrate), sodium salt of metaphosphoric acid (sodium metaphosphate), and sodium salt of polyphosphoric acid (sodium polyphosphate) are particularly preferred.

In the present invention, the best among the tetraacetic acid ethylenediamine salts is disodium edetate hydrate (hereinafter, also simply referred to as "tetraacetic acid ethylenediamine sodium hydrate").

Moreover, these chelating agents can be used individually by 1 type, and can also be used in combination of 2 or more types arbitrarily.

The concentration of the chelating agent in the eye drops is, for example, 0.0001 to 1% (w/v), but 0.0005 to 0.5% (w/v) is preferred, and 0.001 to 0.1% (w/v) is particularly preferred.

In the aqueous eye drops of the present invention, nonionic surfactants can be blended as necessary. The nonionic surfactant is not particularly limited as long as it is within a medically acceptable range, but examples include polyoxyethylene fatty acid esters, polyoxyethylene sorbitan fatty acid esters, and polyoxyethylene castor oil. Derivatives, polyoxyethylene polyoxypropylene glycol, sucrose fatty acid ester, etc. Polyoxyethylene fatty acid esters include polyoxyl 40 stearate, etc., and polyoxyethylene sorbitan fatty acid esters include polysorbate 80 and polysorbate. Alcohol ester 60, polysorbate 40, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan trioleate, polysorbate 65, etc., as far as polyoxyethylene castor oil derivatives are concerned, Examples include polyoxyethylene hardened castor oil 10, polyoxyethylene hardened castor oil 40, polyoxyethylene hardened castor oil 50, polyoxyethylene hardened castor oil 60, polyethylene glycol 5 castor oil, polyethylene glycol 9 castor oil, Polyethylene glycol 15 castor oil, polyethylene glycol 35 castor oil, polyethylene glycol 40 castor oil, etc. For polyoxyethylene polypropylene glycol, polyoxyethylene (160) polyoxypropylene (30) glycol can be mentioned , Polyoxyethylene (42) polyoxypropylene (67) glycol, polyoxyethylene (54) polyoxypropylene (39) glycol, polyoxyethylene (196) polyoxypropylene (67) glycol, polyoxyethylene ( 20) Polyoxypropylene (20) diol and the like.

In the present invention, nonionic surfactants include, for example, polyoxyethylene sorbitan fatty acid esters, preferably polysorbate 80, polysorbate 60, polysorbate 40, and polyoxyethylene sorbitan. Ethylene sorbitan monolaurate, polyoxyethylene sorbitan trioleate, polysorbate 65, etc., particularly preferably polysorbate 80.

Moreover, these nonionic surfactants can be used individually by 1 type, and can also be used in combination of 2 or more types arbitrarily.

The concentration of the non-ionic surfactant in the eye drops is, for example, 0.0001-10% (w/v), but 0.0005-1% (w/v) is preferred, and 0.0005-0.1% (w/v) is Especially good.

In this eyedrop, widely used techniques can be used, and pharmaceutically acceptable additives can be added as necessary. For example, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium acetate, ε-amino group can be selected and added as necessary. Buffers such as caproic acid; isotonizing agents such as sodium chloride, potassium chloride, concentrated glycerin, etc.

The pH of this ophthalmic solution may be within the allowable range of ophthalmic preparations, and it is usually within the range of 4-8. It is suitable to add pH regulators such as hydrochloric acid or sodium hydroxide to this eyedrop.

The eye drops can also be used as soft contact lenses when wearing soft contact lenses. As for soft contact lenses, for example, contact lenses or silicone contact lenses containing hydroxyethyl methacrylate as a main component can be cited.

There are no particular restrictions on the type of the soft contact lens to which this eyedrop is applied, and it may be ionic or non-ionic, hydrated or non-aqueous. For example, in addition to reusable lenses, it can be applied to all soft contact lenses currently on the market or in the future, such as disposable lenses for one day, discarded lenses for one week, and discarded lenses for two weeks.

This eyedrop and the usage of this eyedrop can be appropriately changed according to the dosage form, the severity of the symptoms that should be administered to the patient, age, weight, doctor's judgment, etc. For example, if the eyedrop is selected as the dosage form, it can be divided into 1 It is administered locally 1 to 10 times a day, preferably 2 to 8 times a day, and more preferably 4 to 6 times a day.

According to the definition of dry eye as "a chronic disease caused by various factors in the conjunctiva and epithelium, and accompanied by eye discomfort or visual abnormalities", dry keratoconjunctivitis (KCS) is included in dry eye. In the present invention, the occurrence of dry eye symptoms caused by wearing soft contact lenses is also included in dry eye.

Symptoms of dry eye include not only subjective symptoms such as dry eyes, eye discomfort, eye fatigue, heaviness, photophobia, eye pain, blurred vision, etc., but also hyperemia, corneal and conjunctival epithelium Obstacles, etc. objectively seen.

Regarding the cause of dry eye, there are many unclear points, but there have been reports of Sjogren syndrome; congenital adenopathy; sarcoidosis; transplantation caused by bone marrow transplantation For host disease (GVHD: Graft Versus Host Disease); ophthalmic pemphigus; Stevens-Johnson syndrome (Stevens-Johnson syndrome); trachoma and other causes of occlusion; diabetes; corneal refractive surgery (LASIK: Laser (-assisted) in Situ Keratomileusis (laser-assisted in situ keratomileusis), etc., decreased reflex secretion; meibomian gland dysfunction; decreased oil layer due to blepharitis, etc.; protruding eyes, Insufficient blinking or incomplete eyelid closure caused by rabbit eyes; decreased mucin secretion by embryonic cells; and VDT operation as the cause.

This eye drops can be used for the "prevention and/or treatment of dry eye".

In the present invention, "prevention and/or treatment of dry eye" refers to the prevention and/or treatment or amelioration of the symptoms and/or the definition of symptoms associated with dry eye, and not only refers to the dry eye associated with dry eye Prevention and/or treatment or improvement of subjective symptoms such as sensation, eye discomfort, eye fatigue, heaviness, photophobia, eye pain, blurring vision, etc., including hyperemia and corneal conjunctiva associated with dry eye Prevention and/or treatment or improvement of epithelial disorders, etc. In addition, the "prevention and/or treatment of dry eye" also includes the prevention and/or treatment or improvement of dry eye symptoms by improving the stability of the liquid layer in the eye wearing soft contact lenses. In addition, the prevention and/or treatment or amelioration of dry eye symptoms also means that the prevention and/or treatment or improvement of dry eye symptoms in patients with dry eye who wear soft contact lenses to worsen dry eye symptoms are caused by wearing soft contact lenses Prevention and/or treatment or improvement of the symptoms of dry eye.

In the present invention, the improvement of the stability of the liquid layer means that the liquid is improved in quantity or quality. In addition, the stability of the liquid layer can be confirmed by measuring the BUT (break time of the liquid layer). Among BUTs, in particular, a measurer that is close to a more natural state without the load of staining liquid or the like is called NIBUT (Non-invasive liquid layer destruction time).

In the present invention, "prevention or treatment of eye dryness or eye discomfort in eyes wearing soft contact lenses" means the eye dryness accompanied by the instability of the liquid layer caused by wearing soft contact lenses Or prevention or treatment of eye discomfort or prevention or treatment of eye dryness or eye discomfort caused by corneal and conjunctival epithelial disorders caused by the aforementioned instability.

The present invention provides the preparation of an aqueous ophthalmic solution containing diquafosol or its salt and lohxidine at a concentration of 0.1-10% (w/v) by using 0.005% (w/v) A method for suppressing the reduction of the residual rate of loxidine in the aqueous eye drops with loxidine at the following concentration. Among them, the concentration of loxidine used is 0.005% (w/v) or less, or 0.0001 to 0.005% (w/v) is preferred.

The present invention provides a method for stabilizing loxidine in an aqueous ophthalmic solution containing diquafosol or its salt at a concentration of 0.1-10% (w/v) and loxidine. Among them, it is preferable that the concentration of loxidine is greater than 0.001% (w/v), or greater than 0.001% (w/v) to 0.1% (w/v) or less. [Example]

The following presents a preservation effect test, a deformation inhibition evaluation test for soft contact lenses, a cytotoxicity test using corneal epithelial cells, an evaluation test for the ascending effect of NIBUT, a comparative test for the ascending effect of NIBUT, a test for determining the residual rate of loxidine gluconate, and The results of the long-term stability test of loxidine gluconate, but these examples are for a better understanding of the present invention and do not limit the scope of the present invention. In addition, CL is an abbreviation for contact lens, and SCL is an abbreviation for soft contact lens.

Test Example 1. Preservation effect test Put eye drops 1 to 6 in the prescriptions shown in Table 1, and conduct a preservation effect test.

(Sample preparation) Eye drops 1: Prepare eye drops 1 according to the prescription shown in Table 1. Specifically, diquafosol sodium (3g), sodium hydrogen phosphate hydrate (0.2g), sodium chloride (0.39g), potassium chloride (0.15g), sodium ethylenediamine tetraacetate hydrate (0.01 g) Polysorbate 80 (0.0005g) and loxidine gluconate (0.002g) were dissolved in water to make 100 mL, and pH adjuster was added to make pH 7.2.

Eye drops 2～6: According to the prescription shown in Table 1, eye drops 2 to 6 were prepared in the same manner as eye drops 1.

[Table 1] (Unit: g/100mL) Eye drops 1 Eye drops 2 Eye drops 3 Eye drops 4 Eye drops 5 Eye drops 6 Diquafox Sodium 3 3 3 3 3 3 Sodium hydrogen phosphate hydrate 0.2 0.2 0.2 0.2 0.2 0.2 Sodium chloride 0.39 0.39 0.39 0.39 0.39 0.39 Potassium Chloride 0.15 0.15 0.15 0.15 0.15 0.15 Sodium ethylenediamine tetraacetate hydrate 0.01 0.01 0.01 0.01 0.007 0.01 Polysorbate 80 0.0005 0.0005 0.0005 - - - Loxidine gluconate 0.002 0.0025 0.002 0.0025 0.002 0.002 pH 7.2 7.5 7.8 7.5 7.2 7.2

(experiment method) The preservation effectiveness test was conducted in accordance with the preservation effectiveness test method of the 16th edition of the Japanese Pharmacopoeia. In this experiment, E. coli, P. aeruginosa, S. aureus, C. albicans and A. brasiliensis were used as Test bacteria.

(test results) The test results are shown in Table 2.

[Table 2] Eye drops 1 Eye drops 2 Eye drops 3 Eye drops 4 Eye drops 5 Eye drops 6 Escherichia coli Two weeks ＞4.3 ＞5.6 ＞4.3 ＞5.7 ＞4.6 ＞4.3 4 weeks ＞4.3 ＞5.6 ＞4.3 ＞5.7 ＞4.6 ＞4.3 Pseudomonas aeruginosa Two weeks ＞4.7 ＞5.8 ＞4.7 ＞5.6 ＞4.6 ＞4.7 4 weeks ＞4.7 ＞5.8 ＞4.7 ＞5.6 ＞4.6 ＞4.7 Staphylococcus aureus Two weeks ＞4.9 ＞6.0 ＞4.9 ＞5.6 ＞4.5 ＞4.9 4 weeks ＞4.9 ＞6.0 ＞4.9 ＞5.6 ＞4.5 ＞4.9 Candida albicans Two weeks ＞4.5 ＞5.7 ＞4.5 ＞5.4 ＞4.5 4.5 4 weeks ＞4.5 ＞5.7 ＞4.5 ＞5.4 ＞4.5 ＞4.5 Brazil Aspergillus Two weeks 1.7 1.3 1.7 1.0 0.7 1.4 4 weeks 2.4 1.7 2.5 1.7 0.9 1.5 Judgment (JP) Fit Fit Fit Fit Fit Fit

In addition, the test results in Table 2 use log reduction to indicate how much the number of bacteria during the inspection is reduced compared to the number of bacteria inoculated. For example, in the case of "1", it means the number of bacteria during the inspection. The bacterial count is reduced to 10% of the inoculated bacterial count.

As shown in Table 2, eye drops 1 to 6 show compliance with the preservation efficacy test standards of the Japanese Pharmacopoeia.

(Survey) The above results show that the eye drops have excellent preservation effect.

Test Example 2. Deformation suppression evaluation test of soft contact lenses Use eye drops 4 to study the effect on soft contact lenses.

(Sample preparation) According to the prescription shown in Table 1, eye drops 4 were prepared in the same way as eye drops 1.

(Test method) According to the FDA's contact lens classification shown in Table 3, the current group IV contact lenses (2 weeks ACUVUE ^{( registered trademark )} ) were immersed in eye drops 4 for 24 hours, and the contact lenses before and after were calculated The change amount of the diameter and the base curve shall be examined whether it satisfies the judgment criteria shown in Table 4 below. In addition, the properties of each contact lens after the end of the test were observed. In addition, this judgment criterion is set based on the Ministry of Health, Labour and Welfare Announcement No. 349 Contact Lens Standards for Eyesight Correction (October 5, 2013).

[table 3] Category ¹⁾ nature Group I Water content is less than 50%, non-ionic Group II Water content is less than 50%, non-ionic Group III Water content is above 50%, ionic Group IV Water content is above 50%, ionic ¹⁾ U.S. Food and Drug Administration (FDA) classification

(Judgment criteria)

[Table 4] diameter The tolerance is within ±0.20mm of the displayed diameter. Base curve The tolerance is within ±0.20mm of the displayed basic curve. Traits There are no internal bubbles, impurities, discoloration, or surface damage, and it has a smooth rounded shape.

(result) The results are shown in Table 5.

[table 5] project result Diameter (mm) +0.02 Base curve (mm) +0.02 Traits No change determination Meet the benchmark

As shown in Table 5, the contact lenses after long-term immersion meet the criteria. Accordingly, the eye drops 4 have been shown to suppress the deformation of soft contact lenses.

(Survey) Based on the above results, this ophthalmic solution can be used for soft contact lenses because it inhibits the deformation of soft contact lenses.

Test Example 3. Cytotoxicity test using corneal epithelial cells In order to study the effect of this eye drops on corneal epithelial cells, a cytotoxicity test using corneal epithelial cells was carried out.

(Sample preparation) According to the prescription shown in Table 6, eye drops 7, 8, 9, 10 were prepared in the same manner as eye drops 1.

[Table 6] (Unit: g/100mL) Eye drops 7 Eye drops 8 Eye drops 9 Eye drops 10 Diquafox Sodium 3 - 3 - Sodium hydrogen phosphate hydrate 0.2 0.2 0.2 0.2 Sodium chloride 0.39 0.39 0.41 0.41 Potassium Chloride 0.15 0.15 0.15 0.15 Sodium ethylenediamine tetraacetate hydrate 0.01 0.01 0.01 0.01 Benzalkonium chloride ¹⁾ - - 0.002 0.002 Loxidine gluconate 0.0025 0.0025 - - pH 7.5 7.5 7.5 7.5 ¹⁾ BAK, the chemical structure formula is [C ₆ H ₅ CH ₂ N(CH ₃ ) ₂ C ₁₂ H ₂₅ ]Cl.

(Test method) SV40 immortalized human corneal epithelial cells (HCE-T: Institute of Physics and Chemistry, Biological Resource Center, cell number: RCB2280) were seeded on 96-well plates (1×10 ⁴ cells/well) to contain 10% Cultured in D-MEM/F12 medium of FBS for 1 day. On the next day, the culture medium was exchanged with eyedrop 7, eyedrop 8, eyedrop 9 or eyedrop 10, and the aforementioned corneal epithelial cells were cultured for 15 minutes. Using Cell Proliferation Assay Kit) (manufactured by Promega, catalog number: G3580), the viability of living cells (equivalent to absorbance at 490 nm) was measured.

(result) The test results are shown in Figure 1.

It can be clearly seen from Figure 1 that the diquafoxol eye drops containing loxidine gluconate (eyedrop 7) are better than the eyedrops containing BAK (eyedrop 9, eyedrop 10) and do not contain diquafos Phosphosol or its salt containing loxidine-like eye drops (Eye drops 8) shows higher viable cell activity in cultured immortal human corneal epithelial cells.

(Survey) This eyedrop is because the cultured immortalized human corneal epithelial cells show high viable cell activity. It is safe for living organisms, especially for corneal and conjunctival epithelium. It is not suitable for dry eye and other corneal and conjunctival epithelium. Used in stable diseases.

Test Example 4. Evaluation Test 1 of NIBUT's Ascending Action By wearing soft contact lenses, study the NIBUT of diquafos in the eyes with decreased stability of the liquid layer.

(Sample preparation) According to the prescription shown in Table 1, eye drops 4 were prepared in the same way as eye drops 1.

(Test method) Using a dry eye observation device (DR-1, KOWA) to measure the eye drop 4 (20μL) on the eyes of a cynomolgus monkey ^{wearing soft contact lenses (product name: Menicon Soft MA ( registered trademark ))} /Eye) NIBUT before and 15, 30, 45, and 60 minutes after eye application. Artificial liquid (product name: Soft Santear ^{( registered trademark )} ) was used as a control drug (N=10-11 eyes).

(result) The test results are shown in Figure 2. It can be clearly seen from Fig. 2 that when the eyedrop 4 was instilled into the eye wearing the soft contact lens, a significant increase in NIBUT was observed at all measurement points 60 minutes after the eye was instilled. On the other hand, no increase in NIBUT was observed in artificial dripping eyes.

(Survey) The above results show that the eye drops improve the stability of the liquid layer caused by wearing soft contact lenses. This kind of effect of this eye drops is remarkable even when compared with the artificial liquid that is generally used for the treatment of dry eye. Accordingly, this ophthalmic solution is useful for the prevention and/or treatment of dry eye in eyes wearing soft contact lenses. In addition, the eye drops are also useful for the prevention and/or treatment of eye dryness and/or eye discomfort in eyes wearing soft contact lenses.

Test Example 5. Evaluation test 2 of NIBUT's rising effect By wearing soft contact lenses, study the NIBUT of diquafos in the eyes with decreased stability of the liquid layer.

(Sample preparation) According to the prescription shown in Table 1, eye drops 4 were prepared in the same way as eye drops 1.

(Test method) Measured with a dry eye observation device (DR-1, KOWA) in ^{the eyes of a cynomolgus monkey wearing soft contact lenses (product name: Menicon Soft MA ( registered trademark )} ), and drops eye drops 4 (20μL/eye) NIBUT before and 5, 15, 30, 45, and 60 minutes after eye instillation. Artificial liquid (product name: Soft Santear ^{( registered trademark )} ) and sodium hyaluronate (product name: Hyalein ^{( registered trademark )} Mini eye drops 0.1%) were used as control drugs (N=11 eyes).

(result) The test results are shown in Figure 3. It can be clearly seen from Fig. 3 that when eyedrop 4 was instilled into the eye with soft contact lenses, a significant increase in NIBUT was observed at all measurement points 60 minutes after eye application. On the other hand, no increase in NIBUT was observed during artificial drip infusion. In addition, the increase in NIBUT was observed 5 minutes after the injection of sodium hyaluronate, but the increase was lower than that of the eyedrop 4, and the increase in NIBUT was not observed 15 minutes after the injection.

(Survey) From the above results, it is shown that the eye drops improve the stability of the liquid layer caused by wearing soft contact lenses. This kind of effect of this eyedrop is remarkable even when compared with artificial liquid and sodium hyaluronate eyedrops that are generally used in the treatment of dry eye. Accordingly, this ophthalmic solution is useful for the prevention and/or treatment of dry eye in eyes wearing soft contact lenses. In addition, the eye drops are also useful for the prevention and/or treatment of eye dryness and/or eye discomfort in eyes wearing soft contact lenses.

Test Example 6. Comparison test of NIBUT ascending effect To compare the NIBUT of this eyedrop and eyedrops containing BAK (eyedrops containing diquafosol sodium and BAK) in the eyes with reduced stability of the liquid layer due to wearing soft contact lenses.

(Sample preparation) According to the prescription shown in Table 1, eye drops 4 were prepared in the same way as eye drops 1. In addition, as a comparative example, an eyedrop 11 containing BAK was prepared instead of loxidine gluconate in the eyedrop 4. Specifically, diquafoxol sodium (3g), sodium hydrogen phosphate hydrate (0.2g), sodium chloride (0.41g), potassium chloride (0.15g) and BAK (0.0075g) are dissolved in water to prepare 100 mL, add a pH adjuster to make pH 7.5. Eyedrop 4 and eyedrop 11 are both eyedrops containing the same concentration of active ingredient (diquafoxol sodium). In addition, eyedrops 4 and 11 both meet the preservation efficacy test standards of the Japanese Pharmacopoeia, and are eyedrops with the same preservation effectiveness.

(Test method) Using a dry eye observation device (DR-1, KOWA) to measure 4 points of eye drops in the eyes of cynomolgus monkeys ^{wearing soft contact lenses (product name: Menicon Soft MA ( registered trademark ))} NIBUT (N=11 eyes) before eye drop 11 (20μL/eye) and 30 minutes after eye application.

(result) The test results are shown in Table 7.

[Table 7] Eye drops 4 Eye drops 11 Click the NIBUT in front of you (sec) 3.40 3.39 NIBUT 30 minutes after eye click (sec) 8.20 6.25

The measurement of the original eye drops (eye drops 4) and the eye drops containing BAK (eye drops 11) that meet the preservation efficacy test standards of the Japanese Pharmacopoeia and have the same preservation effect. NIBUT after 30 minutes of eye application is used as The results of the comparative study show that this eyedrop has a higher NIBUT-rising effect than the eyedrop containing BAK.

(Survey) The above results show that this eyedrop is better than eyedrops containing BAK to improve the stability of the liquid layer caused by wearing soft contact lenses.

Test Example 7. Determination Test of Residual Rate of Loxidine Gluconate To study the effect of the concentration of loxidine gluconate on the residual rate of loxidine gluconate after diquafosol ophthalmic solution preparation.

(Sample preparation) Eye drops 12: According to the prescription shown in Table 8, eye drops 12 were prepared. Specifically, it is modulated as follows. 1) Prepare a 0.5% (w/v) loxidine gluconate solution. 2) Prepared containing 0.4% (w/v) sodium hydrogen phosphate hydrate, 0.78% (w/v) sodium chloride, 0.3% (w/v) potassium chloride and 0.02% (w/v) ethylene diacetate Aqueous solution of sodium amine hydrate (2 times thick liquid). 3) Add 1) the prepared solution to water, and further add 2) the prepared solution. 4) After stirring using a magnetic stirrer, add diquafoxone sodium, and further add a pH adjuster (sodium hydroxide or hydrochloric acid) to adjust the pH to 7.5. 5) Adjust the full amount with water so as to have the concentration shown in Table 8. 6) Use a 0.22μm filter to filter and prepare eye drops 12. Eye drops 13, Eye drops 14: The eye drops 13 and the eye drops 14 are prepared in the same manner as the eye drops 12.

[Table 8] (Unit: g/100mL) Eye drops 12 Eye drops 13 Eye drops 14 Diquafox Sodium 3 3 3 Sodium hydrogen phosphate hydrate 0.2 0.2 0.2 Sodium chloride 0.39 0.39 0.39 Potassium Chloride 0.15 0.15 0.15 Sodium ethylenediamine tetraacetate hydrate 0.01 0.01 0.01 Loxidine gluconate 0.0025 0.005 0.01 pH 7.5 7.5 7.5

(experiment method) Measure the content of lohxidine gluconate in the above eye drops 12-14, and calculate the residual rate.

(result) The test results are shown in Table 9.

[Table 9] Eye drops 12 Eye drops 13 Eye drops 14 Survival rate (%) 98.1 97.9 92.4

(Survey) As shown in Table 9, it is clear that in the ophthalmic solution 14 using loxidine gluconate at a concentration of 0.01% (w/v), the residual rate of loxidine gluconate decreased after the eyedrop was prepared . On the other hand, the eye drops 12 and 13 of loxidine gluconate gluconate with concentrations of 0.0025% (w/v) and 0.005% (w/v) remained high even after the eye drops were prepared. The survival rate of loxidine gluconate. As shown above, for the preparation of diquafosol eye drops containing loxidine gluconate, it is better to use loxidine gluconate at a concentration of less than 0.005% (w/v).

Test Example 8. Long-term stability test of loxidine gluconate To study the long-term stability of loxidine gluconate in diquafoxol eye drops.

(Sample preparation) In the same manner as in Test Example 7, eye drops 15-17 were prepared in accordance with the prescriptions shown in Table 10.

[Table 10] Eye drops 15 Eye drops 16 Eye drops 17 Diquafox Sodium 3 3 3 Sodium hydrogen phosphate hydrate 0.2 0.2 0.2 Sodium chloride 0.39 0.39 0.39 Potassium Chloride 0.15 0.15 0.15 Sodium ethylenediamine tetraacetate hydrate 0.01 0.01 0.01 Loxidine gluconate 0.001 0.0025 0.01 pH 7.0 7.0 7.0

(experiment method) The content of loxidine gluconate when the eye drops 15-17 were stored at 40°C/20%RH for 6 months each was quantified using high-speed liquid chromatography (HPLC), and the residual rate (%) was calculated.

(Result) The test result is shown in Table 11. [Table 11] Eye drops 15 Eye drops 16 Eye drops 17 Survival rate (%) 92.8 97.6 100.0

(Survey) Containing 0.0025% (w/v), 0.01% (w/v) lohxidine gluconate eye drops 16, eye drops 17 are more containing 0.001% (w/v) concentration of gluconic acid Loxidine Eye Drop 15 maintains a high residual rate of loxidine gluconate for 6 months at 40℃/20%RH, and shows excellent long-term stability. That is, it is better to stabilize the loxidine gluconate in the diquafosine ophthalmic solution, which shows that the concentration of loxidine gluconate is greater than 0.001% (w/v).

Above, the results of Test Examples 7 and 8 show that it contains a specific concentration range, that is, diquafoxol eye drops containing loxidine at a concentration of 0.001 to 0.005% (w/v). During the preparation of the solution, a high residual rate of hexidine is maintained, and the lohexidine in this ophthalmic solution has excellent long-term stability. Therefore, as shown in Test Example 1, this ophthalmic solution is also expected to have excellent storage efficiency over a long period of time.

[Preparation example] The preparation examples are listed to more specifically illustrate the agent of the present invention, but the present invention is not limited to these preparation examples.

(Prescription example 1: Eye drops (3%(w/v))) In 100mL Ground boasting about phosphorus sodium 3g Sodium hydrogen phosphate hydrate 0.1~0.5g Sodium chloride 0.01~1g Potassium chloride 0.01~1g Sodium ethylenediamine tetraacetic acid hydrate 0.0001～0.1g Lohexidin gluconate 0.0001～0.1g Sterilized pure water Appropriate amount The above-mentioned eye drops can be prepared by adding diquafosol sodium and the above-mentioned other ingredients to sterilized pure water, and by mixing them well.

(Prescription example 2: Eye drops (3%(w/v))) In 100mL Ground boasting about phosphorus sodium 3g Sodium hydrogen phosphate hydrate 0.1~0.5g Sodium chloride 0.01~1g Potassium chloride 0.01~1g Sodium ethylenediamine tetraacetic acid hydrate 0.0001～0.1g Lohexidin gluconate 0.0001～0.1g Polysorbate 80 0.0001～0.1g Sterilized pure water Appropriate amount The above-mentioned eye drops can be prepared by adding diquafosol sodium and the above-mentioned other ingredients to sterilized pure water, and by mixing them well. [Possibility of Industrial Use]

This eyedrop has excellent preservation effect. Furthermore, this eye drops can be used as soft contact lenses because it inhibits the deformation of soft contact lenses. In addition, compared with the eyedrops containing BAK and the eyedrops containing no diquafosol or its salt and containing lohexidine, this eyedrop system showed higher activity in cultured immortalized human corneal epithelial cells. Cell viability. Therefore, this ophthalmic solution is safer for living bodies, especially for corneal and conjunctival epithelium, and is useful for diseases where corneal and conjunctival epithelium is unstable such as dry eye. In addition, the eye drops that are attached to the eyes wearing soft contact lenses significantly increase NIBUT. On the other hand, such an effect has not been observed with artificial trickling liquid. That is, the eye drops are attached to the eyes wearing soft contact lenses to stabilize the liquid layer. The occurrence/exacerbation of dry eye symptoms caused by wearing soft contact lenses is due to the decrease in the stability of the liquid layer. The stabilization of the liquid layer caused by the eye drops is useful for wearing soft contact lenses. Prevention and/or treatment of dry eye in your eyes. In addition, the eye drops are also useful for the prevention and/or treatment of eye dryness and/or eye discomfort in eyes wearing soft contact lenses. In addition, the diquafosine ophthalmic solution containing lohxidine in a specific concentration range, that is, 0.001-0.005% (w/v) concentration, maintains high lohxi during the preparation of the eye drops. The residual rate of the fixed class, and the loxidine in this eyedrop has excellent long-term stability.

without.

[Fig. 1] Fig. 1 is a graph showing the results of a cytotoxicity test using corneal epithelial cells in Test Example 3. [Fig. 2] Fig. 2 is a graph showing the results of the evaluation test 1 of the ascending effect of NIBUT in Test Example 4. [Fig. [Fig. 3] Fig. 3 is a graph showing the results of the evaluation test 2 of the rising effect of NIBUT in Test Example 5.

without.

Claims (1)

A method for stabilizing chlorhexidine in an aqueous eyedrop containing diquafosol or its salt at a concentration of 0.1-10% (w/v) , And loxidine, the concentration of the loxidine is greater than 0.001% (w/v) to less than 0.01% (w/v).

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