Anti-acanthamoeba liquid formulation

Abstract

The purpose of the present invention is to provide a versatile anti-acanthamoeba liquid formulation that, while having an anti-acanthamoeba function, has less impact on ocular tissues, is safe, and can be applied to a wide range of contact lenses. The purpose is solved by an anti-acanthamoeba liquid formulation or the like that contains sodium dihydrogen phosphate, a basic isotonizing agent, and a poly diallyldimethylammonium chloride having an average molecular weight of 10,000-1,100,000.

Description

Anti-Acanthamoeba liquid

[0001] The present invention relates to a liquid preparation having a disinfecting effect against Acanthamoeba.

[0002] Acanthamoeba corneal infection is a corneal infection caused by the infection of the cornea by a microorganism of the genus Acanthamoeba (hereinafter simply referred to as Acanthamoeba). Patients with Acanthamoeba corneal infection not only experience severe eye pain, but in severe cases, the cornea may develop holes, leading to blindness. The number of cases of Acanthamoeba corneal infection has been increasing in recent years.

[0003] Acanthamoeba is a protozoan found widely in soil and freshwater, including tap water. Normally, Acanthamoeba is unlikely to cause infection in a healthy cornea. However, if the cornea has a wound, Acanthamoeba can invade and attach to the wound, causing infection. Contact lens wearers can develop Acanthamoeba corneal infection by wearing contact lenses contaminated with Acanthamoeba from tap water or other sources.

[0004] Acanthamoeba corneal infection is generally treated by corneal scraping and administration of antifungal drugs. However, Acanthamoeba corneal infection is an intractable infection that places a heavy burden on patients, posing problems such as difficulty in early detection, a tendency for inflammation to recur, prolonged treatment periods, and corneal scarring. Contact lens wearers can prevent or treat corneal infections by using a liquid containing a drug with anti-Acanthamoeba activity that can prevent contamination by Acanthamoeba.

[0005] Known examples of such liquid preparations include an anti-Acanthamoeba disinfectant and preservative for contact lenses containing an effective amount of polylysine (see, for example, Patent Document 1); an anti-Acanthamoeba composition containing, as an active ingredient, lactoferrin, a glycoprotein, or lactoferricin, a peptide produced by enzymatic degradation of lactoferrin (see, for example, Patent Document 2); and an antiprotozoan composition containing, as active ingredients, the candin drug caspofungin and a biguanide compound (see, for example, Patent Document 3).

[0006] Meanwhile, in recent years, multi-purpose solutions (MPS), which can clean, rinse, disinfect, and store contact lenses with a single solution, have become widely used due to their simple processing. Organic nitrogen-based disinfectants are commonly used as disinfectant components in MPS. Known as an agent applicable to MPS is a preventive and therapeutic agent for Acanthamoeba keratitis, which contains two active ingredients: an organic nitrogen-based disinfectant and a diallyldialkylammonium-containing polymer (see, for example, Patent Document 4). Also known is a method of enhancing the bactericidal activity against Acanthamoeba by combining alexidine with a polyhydric alcohol and boric acid (see, for example, Patent Document 5).

[0007] JP 2002-143277 A JP 2011-246458 A JP 2013-234176 A JP 2014-218461 A JP 2018-035151 A

[0008] However, the agents, compositions, and methods described in Patent Documents 1 to 5 each have problems. The anti-Acanthamoeba disinfectant and preservative described in Patent Document 1 has a problem of poor versatility because the active ingredient, polylysine, cannot be applied to Group IV (ionic high water content) soft contact lenses.

[0009] The anti-Acanthamoeba composition described in Patent Document 2 has the problem that the active ingredient, glycoprotein or its degradation peptide, deposits in ocular tissue, causing visual impairment and eye damage, and is therefore low in safety.

[0010] The antiprotozoan composition described in Patent Document 3 exerts antifungal activity by inhibiting the activity of 1,3-β-glucan synthase, a major component of cell walls, using the active ingredient caposfungin. However, caposfungin causes side effects such as anaphylaxis, liver dysfunction, and eye pruritus, making it unsafe.

[0011] The preventive and therapeutic agent for Acanthamoeba keratitis described in Patent Document 4 contains a high concentration of a diallyldialkylammonium-containing polymer as an active ingredient, but there is a problem in that the polymer may cause side effects such as anaphylaxis, liver dysfunction, and ocular pruritus.

[0012] The method described in Patent Document 5 is characterized by combining alexidine with a polyhydric alcohol and boric acid. Of these, boric acid has been widely used in ophthalmic solutions due to its antiseptic effect. However, in recent years, it has been reported that boric acid can damage the cornea, resulting in eye disorders, and therefore the incorporation of boric acid, particularly in ophthalmic solutions, has begun to be restricted.

[0013] As described above, even though the active ingredients of the agents, compositions, and methods described in Patent Documents 1 to 5 exhibit disinfecting effects against Acanthamoeba, there are problems with safety to ocular tissues and versatility of the contact lenses to which they are applied.

[0014] Therefore, an object of the present invention is to provide an anti-Acanthamoeba liquid preparation that has anti-Acanthamoeba activity, has little effect on ocular tissue, is safe, and is versatile enough to be applied to a wide range of contact lenses.

[0015] In order to solve the above problems, the present inventors have conducted extensive research into a liquid preparation that has a disinfecting effect against Acanthamoeba while having little effect on ocular tissue.In the research, a combination of sodium dihydrogen phosphate and a basic isotonic agent, both of which have been proven to be safe for ocular tissue, was used, and a disinfecting effect against fungi was confirmed, but no disinfecting effect against Acanthamoeba was observed.

[0016] Therefore, the inventors further repeated trial and error and, as a result, focused on polydiallyldimethylammonium chloride, which has also been confirmed to be safe for ocular tissue, and discovered that by using a specific polydiallyldimethylammonium chloride in combination with sodium dihydrogen phosphate and a basic isotonic agent, an excellent disinfecting effect against Acanthamoeba was exhibited.

[0017] Surprisingly, the solution containing sodium dihydrogen phosphate, a basic tonicity agent, and a selected polydiallyldimethylammonium chloride was found to be compatible with a wide variety of contact lenses.

[0018] Based on these findings, the present inventors have finally succeeded in creating a liquid preparation containing sodium dihydrogen phosphate, a basic tonicity agent, and a specific polydiallyldimethylammonium chloride, which can solve the problems of the present invention. The present invention has been completed based on the findings first discovered by the present inventors and the successful examples achieved.

[0019] Therefore, according to the present invention, there are provided anti-Acanthamoeba liquid preparations of the following embodiments: [1] An anti-Acanthamoeba liquid preparation comprising sodium dihydrogen phosphate, a basic tonicity agent, and polydiallyldimethylammonium chloride having an average molecular weight of 10,000 to 1,100,000. [2] The anti-Acanthamoeba liquid preparation according to item [1], wherein the ratio of the content of the basic tonicity agent to the content of the sodium dihydrogen phosphate ([basic tonicity agent] / [sodium dihydrogen phosphate]) is 0.01 to 2 and the pH is 6.5 to 8.5. [3] The anti-Acanthamoeba liquid preparation according to item [1] or [2], wherein the ratio of the content of the diallyldimethylammonium chloride to the content of the sodium dihydrogen phosphate ([polydiallyldimethylammonium chloride] / [sodium dihydrogen phosphate]) is 0.00005 to 0.01. [4] The anti-Acanthamoeba liquid preparation according to any one of items [1] to [3], wherein the ratio of the content of the polydiallyldimethylammonium chloride to the total content of the sodium dihydrogen phosphate and the basic tonicity agent ([polydiallyldimethylammonium chloride] / ([sodium dihydrogen phosphate]+[basic tonicity agent]) is 0.000001 to 1. [5] The anti-Acanthamoeba liquid preparation according to any one of items [1] to [4], wherein the basic tonicity agent is one or more basic tonicity agents selected from the group consisting of 2-amino-2-hydroxymethyl-1,3-propanediol, 2-amino-2-ethyl-1,3-propanediol, arginine, and salts thereof. [6] The anti-Acanthamoeba liquid preparation according to any one of items [1] to [5], wherein the polydiallyldimethylammonium chloride has an average molecular weight of 100,000 to 1,000,000. [7] The anti-Acanthamoeba liquid preparation according to items [1] to [6], wherein the liquid preparation further contains an isotonicity agent other than the basic isotonicity agent. [8] The anti-Acanthamoeba liquid preparation according to item [7], wherein the isotonicity agent is one or more isotonicity agents selected from the group consisting of sodium chloride, potassium chloride, 1,2-propanediol, 1,3-propanediol, mannitol, and sorbitol.[9] The anti-Acanthamoeba liquid preparation according to any one of items [1] to [8], further comprising one or more quaternary ammonium salts selected from the group consisting of polyhexanide hydrochloride, alexidine, myristamidopropyldimethylamine, benzalkonium chloride, benzethonium chloride, and polidronium chloride.

[0020] According to the present invention, Acanthamoeba can be killed by a combination of ingredients whose safety has been proven in ophthalmic solutions. Furthermore, according to the present invention, a wide variety of contact lenses can be disinfected against Acanthamoeba without causing deformation or the like. Thus, the anti-Acanthamoeba solution of one embodiment of the present invention is highly safe for ocular tissues, has versatility in that it can be applied to a wide range of contact lenses, and yet exhibits excellent disinfecting effects against Acanthamoeba.

[0021] Furthermore, the anti-Acanthamoeba solution of one embodiment of the present invention can be used for cleaning, rinsing, disinfecting, and storing contact lenses, and can therefore be used routinely as an MPS. According to the present invention, contact lens wearers can reduce or avoid the risk of contracting Acanthamoeba corneal infection caused by Acanthamoeba.

[0022] Each aspect of the present invention will be described in detail below, but the present invention can take various forms as long as the object of the present invention is achieved.

[0023] Unless otherwise specified, the terms used in this specification are used in the sense commonly used by those skilled in the art of chemicals dealing with ophthalmic lenses such as contact lenses, and should not be construed as having an unduly limiting meaning. Furthermore, the speculations and theories made in this specification are based on the inventors' knowledge and experience to date, and therefore the present invention is not limited solely to such speculations and theories.

[0024] "Content" is synonymous with concentration and amount used (added amount) and refers to the ratio of the amount of a component to the total amount of the final product. However, the total amount of the component content does not exceed 100%. In this specification, unless otherwise specified, the unit of content is "mass by volume % (w / v%)." When using a commercially available product, the content of a component is preferably the amount of the component contained in the commercially available product, but it may also be the amount of the commercially available product itself. The term "and / or" refers to any one of the listed related items, or any combination of two or more of the listed items, or all combinations. "Comprise" means that elements other than those explicitly stated as included can be added (synonymous with "comprise at least"), but also encompasses "consist" and "essentially consist." In other words, "comprise" can mean including the specified elements and any one or more elements, consisting of the specified elements, or consisting essentially of the specified elements. Elements include limitations such as components, steps, conditions, parameters, etc. The "to" in a numerical range includes the preceding and following numerical values, and also includes ranges excluding one of the included limits. For example, "0 w / v % to 100 w / v %" means 0 w / v % or more, 100 w / v % or less, or 0 w / v % or more and 100 w / v % or less. The number of digits in an integer value matches the number of significant figures. For example, 1 has one significant digit, and 10 has two significant digits. Furthermore, for decimal values, the number of digits after the decimal point matches the number of significant digits. For example, 0.1 has one significant digit, and 0.10 has two significant digits.

[0025] [Effect of Anti-Acanthamoeba Liquid Preparation] The anti-Acanthamoeba liquid preparation of one embodiment of the present invention contains a combination of sodium dihydrogen phosphate, a basic isotonic agent, and polydiallyldimethylammonium chloride having a predetermined average molecular weight, and thereby has the effect of killing, removing, and / or inhibiting the growth of Acanthamoeba (anti-Acanthamoeba effect). In this specification, killing, removing, or inhibiting the growth of microorganisms may also be referred to as "disinfection" or "sterilization."

[0026] The anti-Acanthamoeba liquid preparation of one embodiment of the present invention has an anti-Acanthamoeba activity against microorganisms of the genus Acanthamoeba. The level of the anti-Acanthamoeba activity can be confirmed using Acanthamoeba castellanii ATCC 50370 strain as an indicator. The anti-Acanthamoeba activity of the anti-Acanthamoeba liquid preparation of one embodiment of the present invention may be such that a disinfecting effect is observed against the Acanthamoeba castellanii ATCC 50370 strain. For example, based on the method described in the Examples below, the dead amoebae rate is preferably 80% or more, and more preferably 90% or more, according to Evaluation Method 1 for Acanthamoeba disinfection effect, and / or is such that the logarithmic difference is evaluated to be 0.5 or more, more preferably 1.0 or more, even more preferably 1.5 or more, and even more preferably 2.0 or more, according to Evaluation Method 2 for Acanthamoeba disinfection effect.

[0027] The anti-Acanthamoeba liquid preparation of one embodiment of the present invention is highly safe to ocular tissue. The safety need only be such that it does not have any significant negative effect on ocular tissue. For example, when evaluated based on the safety evaluation method described in the Examples below, the relative colony formation rate is preferably 75% or more when the concentration is adjusted to 2.5 v / v%, more preferably 75% or more when the concentration is adjusted to 5 v / v%, even more preferably 75% or more when the concentration is adjusted to 10 v / v% or more, and even more preferably 75% or more when the concentration is adjusted to 20 v / v%.

[0028] The anti-Acanthamoeba liquid preparation of the present invention has contact lens compatibility with a wide variety of contact lenses, such that it does not change the power, base curve, diameter, etc. Contact lens compatibility may be such that it does not cause any significant change in the shape of the contact lens to which it is applied. For example, when evaluated based on the contact lens compatibility evaluation described in the Examples below, the change in power is preferably within ±0.25 D, and / or the change in base curve (BC) is preferably within ±0.2 mm, and / or the change in diameter (DIA) is preferably within ±0.2 mm.

[0029] [Active ingredient] An anti-Acanthamoeba liquid preparation according to one embodiment of the present invention contains sodium dihydrogen phosphate, a basic isotonic agent, and polydiallyldimethylammonium chloride (hereinafter simply referred to as polydiallyldimethylammonium chloride) having a predetermined average molecular weight as active ingredients for exerting an anti-Acanthamoeba effect.

[0030] Sodium dihydrogen phosphate has the chemical formula NaH ２ P.O. ４ As long as it is one of the inorganic phosphate compounds represented by the formula (I), it may be an anhydrous compound or a hydrate such as a monohydrate or dihydrate, but the commonly used sodium dihydrogen phosphate dihydrate is preferred. Furthermore, an anti-Acanthamoeba liquid preparation according to one embodiment of the present invention may contain another inorganic phosphate compound in addition to sodium dihydrogen phosphate. Examples of such inorganic phosphate compounds include disodium hydrogen phosphate, trisodium phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, tripotassium phosphate, calcium dihydrogen phosphate, and the like, which may be an anhydrous compound or a hydrate. The other inorganic phosphate compound may be any one of the above, or two or more may be used in combination.

[0031] The content of sodium dihydrogen phosphate may be any amount that exhibits an anti-Acanthamoeba effect when combined with a basic tonicity agent and polydiallyldimethylammonium chloride, and may be set appropriately, for example, preferably 0.001 w / v% to 5 w / v%, more preferably 0.01 w / v% to 3 w / v%, even more preferably 0.05 w / v% to 2 w / v%, and even more preferably 0.1 w / v% to 1 w / v%. The total content of sodium dihydrogen phosphate and the content of another inorganic phosphate compound may be set appropriately, for example, preferably 0.01 w / v% to 5 w / v%, more preferably 0.05 w / v% to 3 w / v%, and even more preferably 0.1 w / v% to 2 w / v%.

[0032] The basic tonicity agent refers to an agent that has a basic group and is used to adjust the osmotic pressure of the anti-Acanthamoeba liquid preparation of one embodiment of the present invention to a desired range (e.g., similar to that of tears). Examples of basic tonicity agents include ammonia, monoethanolamine, diethanolamine, triethanolamine, 2-amino-2-methyl-1-propanol, 2-amino-2-methyl-1,3-propanediol, 2-amino-2-hydroxymethyl-1,3-propanediol, 2-amino-2-ethyl-1,3-propanediol, arginine, and salts thereof. However, from the viewpoint of having little effect on ocular tissue and exhibiting a buffering effect at a pH near neutral, organic buffers having an amino group are preferred, and 2-amino-2-hydroxymethyl-1,3-propanediol, 2-amino-2-ethyl-1,3-propanediol, and arginine are more preferred. The basic tonicity agent may be any one of these alone or in combination of two or more. The basic tonicity agent exhibits a strong buffering effect at near-neutral pH, and is therefore preferably 2-amino-2-hydroxymethyl-1,3-propanediol or a combination of 2-amino-2-hydroxymethyl-1,3-propanediol with another basic tonicity agent.

[0033] The content of the basic isotonic agent may be any amount that exhibits an anti-Acanthamoeba effect when combined with sodium dihydrogen phosphate and diallyldimethylammonium polychloride, and can be appropriately set taking into consideration irritation to ocular tissues and osmotic pressure; for example, it is preferably 0.001 w / v% to 20 w / v%, more preferably 0.005 w / v% to 10 w / v%, and even more preferably 0.008 w / v% to 5 w / v%.

[0034] From the viewpoint of anti-Acanthamoeba activity, it is preferable that the ratio of the content of sodium dihydrogen phosphate to the content of the basic tonicity agent be within a predetermined range. The ratio of the content of the basic tonicity agent to the content of sodium dihydrogen phosphate ([basic tonicity agent] / [sodium dihydrogen phosphate]) is, for example, preferably 0.001 to 10, more preferably 0.005 to 5, even more preferably 0.01 to 3, and still more preferably 0.04 to 2. The total content of sodium dihydrogen phosphate and the basic tonicity agent may be appropriately set taking into consideration the irritation to ocular tissues and the osmotic pressure of the anti-Acanthamoeba liquid preparation of one embodiment of the present invention.

[0035] Polydiallyldimethylammonium chloride is a cationic polymer having a predetermined average molecular weight and containing diallyldimethylammonium chloride units in the molecule, as shown in the following general formula (I): Polydiallyldimethylammonium chloride is also known as polyquaternium-6. [In the formula, n represents an integer set so that the average molecular weight of the polymer falls within the range specified above.]

[0036] The polydiallyldimethylammonium chloride has a predetermined average molecular weight, specifically 10,000 to 1,100,000, but in consideration of safety to ocular tissues while still exhibiting an anti-Acanthamoeba activity, for example, polydiallyldimethylammonium chloride having an average molecular weight in the range of preferably 100,000 to 1,000,000, more preferably 120,000 to 800,000, and even more preferably 150,000 to 500,000. The average molecular weight represents the weight-average molecular weight.

[0037] The method for producing polydiallyldimethylammonium chloride is not particularly limited, but examples thereof include a method in which an aqueous solution of diallyldimethylammonium chloride is used as a raw material, and a linear homopolymer is obtained by cyclopolymerization using a radical polymerization initiator such as a peroxide or by light irradiation.

[0038] Commercially available polydiallyldimethylammonium chloride products may be used, such as "MERQUAT100" (molecular weight: 150,000) from Lubrizol, "ME Polymer H-40W" (molecular weight: 240,000) from Toho Chemical Industry Co., Ltd., "Cosmot VG" (molecular weight: 200,000) from Senka Corporation, "Unisense FPA1002L" (molecular weight: 500,000) from Senka Corporation, "Unisense FPA7000E" (molecular weight: 1,200,000) from Senka Corporation, "Flocare C106MV" (molecular weight: 150,000) from SNF Corporation, and "Genamin PDAC" (molecular weight: 230,000) from Clariant International (the polymer names are registered trademarks or product names, and the same applies hereinafter to polymers). Furthermore, polydiallyldimethylammonium chloride may be polydimethyldimethylenepyrrolidinium chloride solution in accordance with Pharmaceutical and Quasi-drug Raw Materials Standards 2021 (External Standards).

[0039] The content of diallyldimethylammonium polychloride can be set as appropriate as long as it is an amount that exhibits an anti-Acanthamoeba effect when combined with sodium dihydrogen phosphate and a basic isotonic agent. However, considering that diallyldimethylammonium polychloride has a significant effect on the disinfecting effect, for example, the content is preferably 0.00001 w / v% to 10 w / v%, more preferably 0.00005 w / v% to 1 w / v%, and even more preferably 0.00008 w / v% to 0.05 w / v%.

[0040] In order for the combination of sodium dihydrogen phosphate, a basic tonicity agent, and polydiallyldimethylammonium chloride to function as an active ingredient, the total content of these components is preferably within a predetermined range, for example, preferably 0.01 w / v % to 10 w / v %, more preferably 0.05 w / v % to 5 w / v %, and even more preferably 0.1 w / v % to 3 w / v %.

[0041] From the viewpoint of anti-Acanthamoeba activity, it is preferable that the ratio of the content of polydiallyldimethylammonium chloride to the content of sodium dihydrogen phosphate or the ratio of the total content of sodium dihydrogen phosphate and the content of the basic isotonic agent is within a predetermined range.

[0042] The ratio of the content of polydiallyldimethylammonium chloride to the content of sodium dihydrogen phosphate ([polydiallyldimethylammonium chloride] / [sodium dihydrogen phosphate]) is preferably 0.00001 to 1, more preferably 0.00005 to 0.5, and even more preferably 0.0001 to 0.1.

[0043] The ratio of the content of polydiallyldimethylammonium chloride to the total content of sodium dihydrogen phosphate and the content of the basic tonicity agent ([polydiallyldimethylammonium chloride] / ([sodium dihydrogen phosphate]+[basic tonicity agent]) is, for example, preferably 0.000001 to 1, more preferably 0.00001 to 0.5, and even more preferably 0.00003 to 0.1.

[0044] When the ratio of the content of polydiallyldimethylammonium chloride to the content of sodium dihydrogen phosphate or the ratio of the total content of sodium dihydrogen phosphate and the content of a basic isotonic agent is within the above range, the anti-Acanthamoeba liquid preparation of one embodiment of the present invention can exhibit a disinfecting effect against Acanthamoeba while also being safe for ocular tissue.

[0045] [Other Components] The anti-Acanthamoeba liquid preparation of one embodiment of the present invention contains sodium dihydrogen phosphate, a basic tonicity agent, and polydiallyldimethylammonium chloride, and may contain other components in addition to these. Examples of other components include isotonicity agents other than basic isotonicity agents. In this specification, when simply referring to an isotonicity agent, it refers to an isotonicity agent other than a basic isotonicity agent, i.e., an isotonicity agent that does not have a basic group.

[0046] The isotonic agent may be any agent that does not have a basic group, and examples thereof include sodium chloride, potassium chloride, propylene glycol (PG), 1,2-propanediol, 1,3-propanediol, mannitol, sorbitol, sodium carbonate, sodium bicarbonate, and glycerin. Any one of these may be used alone, or two or more may be used in combination.

[0047] The content of the isotonic agent may be any amount that allows the anti-Acanthamoeba liquid preparation of one embodiment of the present invention to have an osmotic pressure that is suitable for ocular tissue, and can be set appropriately. For example, the content is preferably 0 w / v % to 10 w / v %, more preferably 0.001 w / v % to 5 w / v %, even more preferably 0.01 w / v % to 3 w / v %, and even more preferably 0.05 w / v % to 2 w / v % or 0.1 w / v % to 1.8 w / v %.

[0048] The other components are not particularly limited as long as they do not interfere with the solution of the problems of the present invention, and examples thereof include additives typically contained in contact lens care products. Examples of additives include disinfectants, surfactants, chelating agents, thickeners, humectants, isotonicity agents, enzymes such as protease and lipase. Some of the other components are listed below, but these are merely examples and are not intended to be limiting.

[0049] Specific examples of disinfectants include biguanide disinfectants such as polyhexamethylene biguanide (PHMB), polyaminopropyl biguanide, alexidine, and chlorhexidine gluconate (GCH); quaternary ammonium disinfectants such as myristamidopropyl dimethylamine, benzalkonium chloride, benzethonium chloride, and polidronium chloride; iodine disinfectants such as chlorhexidine gluconate and povidone-iodine; and hydrogen peroxide disinfectants, preferably organic nitrogen disinfectants, more preferably polyhexamethylene biguanide (PHMB). The content of the disinfectant can be set as appropriate, but is preferably 0 w / v % to 1.0 w / v %, taking into consideration, for example, irritation to ocular tissues and safety.

[0050] Considering the influence on contact lenses, preferred specific examples of surfactants include nonionic surfactants. Specific examples of nonionic surfactants include polyoxyethylene polyoxypropylene glycol, ethylenediamine tetrapolyoxypropylene condensates, polyoxyethylene alkyl ethers, polyoxyethylene alkylphenyl ethers, polyoxyethylene hydrogenated castor oil, polyoxyalkyl esters, polyoxyethylene sulbitan alkyl esters, etc., with polyoxyethylene polyoxypropylene glycol, ethylenediamine tetrapolyoxypropylene condensates and polyoxyethylene hydrogenated castor oil being preferred.

[0051] Specific examples of polyoxyethylene polyoxypropylene glycols include polyoxyethylene (50) polyoxypropylene (40) glycol, polyoxyethylene (300) polyoxypropylene (55) glycol, polyoxyethylene (200) polyoxypropylene (40) glycol, polyoxyethylene (160) polyoxypropylene (30) glycol, polyoxyethylene (196) polyoxypropylene (67) glycol, and polyoxyethylene (200) polyoxypropylene (70) glycol. These are commercially available as Corifol P188, Corifol P407 (manufactured by BASF), and Pluronic (registered trademark) F-68, F-88, F-108, F-77, F-127, L-64, P-84, and P-85 (manufactured by ADEKA Corporation).

[0052] Specific examples of ethylenediamine tetrapolyoxypropylene condensates include tetrafunctional block copolymers of polyoxyethylene polyoxypropylene copolymer and ethylenediamine, which are commercially available as Tetoronic 904, 908, 1104, 1107, 1304, 1504 (manufactured by BASF), Synperonic T304, 707, 908 (manufactured by CRODA), Pluronic (registered trademark) TR-704, TR-913R (manufactured by ADEKA), etc.

[0053] Specific examples of polyoxyethylene hydrogenated castor oil include polyoxyethylene hydrogenated castor oil 40 and polyoxyethylene hydrogenated castor oil 60, which are commercially available as Corifol RH-40 (manufactured by BASF), HCO-40, HCO-50, and HCO-60 (manufactured by Nikko Chemical Co., Ltd.), etc.

[0054] The surfactant may be any one of those listed above, or a combination of two or more thereof. The content of the surfactant can be appropriately determined, but considering, for example, safety to ocular tissues and compatibility with contact lenses, the content is preferably 0 w / v % to 5 w / v %, more preferably 0 w / v % to 2.5 w / v %, even more preferably 0 w / v % to 1 w / v %, and still more preferably 0 w / v % to 0.5 w / v %.

[0055] Specific examples of chelating agents include those typically used in ophthalmic solutions, such as edetic acid, sodium edetate, disodium edetate, ethylenediaminetetraacetic acid, polycarboxylic acids such as citric acid and tartaric acid, and salts thereof, and these can be used alone or in combination of two or more. The content of the chelating agent may be any amount typically used in ophthalmic solutions and can be appropriately set, but, for example, in consideration of safety to ocular tissues, it is preferably 0 w / v% to 10 w / v%, more preferably 0.001 w / v% to 5 w / v%, and even more preferably 0.01 to 1 w / v%.

[0056] Specific examples of thickeners include those typically used in ophthalmic solutions, such as cellulose-based compounds such as hydroxyethyl cellulose, hydroxypropyl methylcellulose (TC-5R, etc.), and sodium carboxycellulose, and vinyl-based compounds such as polyvinyl alcohol and carboxyvinyl polymer. The content of the thickener may be any amount typically used in ophthalmic solutions and can be set appropriately, but in consideration of safety to ocular tissues, the content is preferably 0 w / v% to 5 w / v%, more preferably 0.001 w / v% to 2 w / v%, and even more preferably 0.01 w / v% to 1 w / v%, for example.

[0057] Specific examples of wetting agents include those commonly used in ophthalmic solutions, such as hyaluronic acid and its salts, amino acids and their salts, polyvinyl alcohol, and 2-methacryloyloxyethyl phosphorylcholine polymer.

[0058] An anti-Acanthamoeba liquid preparation according to one embodiment of the present invention is prepared by dissolving each component in a solvent such as water. The solvent is preferably purified water, ion-exchanged water, or other water typically used in ophthalmic liquid preparations. The water content is preferably, for example, 90 w / v % to 99.9 w / v %, and more preferably 95 w / v % to 99.5 w / v %.

[0059] [Anti-Acanthamoeba liquid preparation] The anti-Acanthamoeba liquid preparation of one embodiment of the present invention may have various properties as long as they do not interfere with the solution of the problems of the present invention. For example, when the anti-Acanthamoeba liquid preparation of one embodiment of the present invention is used so as to come into contact with ocular tissue and a contact lens, it is preferable that the anti-Acanthamoeba liquid preparation have properties that do not have a negative effect on the ocular tissue and the contact lens.

[0060] In such cases, in order to reduce irritation to ocular tissue and to enable the basic tonicity agent to function as an isotonicity agent, the anti-Acanthamoeba liquid preparation of one embodiment of the present invention preferably has a pH of 5.5 to 9.5, more preferably 6.0 to 9.0, and even more preferably 6.5 to 8.5; and the osmotic pressure is preferably 200 mOsm / L to 400 mOsm / L, and more preferably 220 mOsm / L to 350 mOsm / L.

[0061] The anti-Acanthamoeba solution of one embodiment of the present invention may be produced according to a method for producing a solution used in the field of ophthalmology known to those skilled in the art. For example, it can be produced by adding sodium dihydrogen phosphate, a basic tonicity agent, and polydiallyldimethylammonium chloride, and optionally other ingredients, simultaneously or sequentially to purified water and mixing them. The anti-Acanthamoeba solution of one embodiment of the present invention may be subjected to filtration or other treatment to remove insoluble components, or may be subjected to sterilization or other treatment to prevent microbial contamination. The anti-Acanthamoeba solution of one embodiment of the present invention is preferably a filtered and / or sterilized solution. Specific examples of the method for producing the anti-Acanthamoeba solution of one embodiment of the present invention can be found in the Examples described below.

[0062] The anti-Acanthamoeba liquid preparation according to one embodiment of the present invention can be used as a liquid composition in the form of a pharmaceutical or quasi-drug preparation, such as eye drops, artificial tears, eyewashes, preservatives, cleaners, and disinfectants for contact lenses.

[0063] The anti-Acanthamoeba liquid preparation of one embodiment of the present invention is preferably a contact lens care solution because it is safe for ocular tissues and compatible with a wide range of contact lenses. When the anti-Acanthamoeba liquid preparation of one embodiment of the present invention is a contact lens care solution, it can be used to clean, rinse, and disinfect contact lenses, and can also be used to store contact lenses by placing the anti-Acanthamoeba liquid preparation of one embodiment of the present invention and a contact lens in a storage container.

[0064] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples, and the present invention can take various forms as long as the object of the present invention can be achieved.

[0065] 1. Preparation of test liquid preparations Test liquid preparations of Examples and Comparative Examples were prepared according to the formulations shown in Tables 1A to 1C. The unit of component amounts in the tables is "w / v %."

[0066] Specifically, each component listed in Tables 1A to 1C was weighed out in the amount listed in Tables 1A to 1C. Next, each component was added to purified water and dissolved, and the resulting solution was diluted to a total volume of 100 mL, followed by thorough stirring to ensure uniformity. The resulting solution was sterilized by filtration using a PES membrane filter with a pore size of 0.2 μm, to prepare each test liquid formulation.

[0067] 2. Method 1 for evaluating disinfection effect against Acanthamoeba Acanthamoeba castellanii (ATCC 50370) was used as the test strain.

[0068] After centrifugation of the culture medium of Acanthamoeba cultured at 28°C in a liquid medium, the precipitated cells were suspended in 1 / 4 Ringer's solution to obtain an Acanthamoeba concentration of 1 x 10 ７ cells / mL~10 ８The concentration was adjusted to 100 cells / mL to obtain an Acanthamoeba suspension. 0.1 mL of the Acanthamoeba suspension was inoculated into 10 mL of each test liquid preparation of the Examples and Comparative Examples. After inoculation, the test liquid preparations were disinfected by incubation at 25°C for 4 hours.

[0069] After the disinfection treatment, the test solution was mixed with 0.4 w / v% Trypan Blue Solution. After mixing, the test solution was counted under an optical microscope using a hemocytometer to measure the total cell count, live cell count, and dead cell count. Based on the measured values, the cell death rate was calculated as the ratio of dead cells to the total cell count. Test solutions that were found to have a calculated cell death rate of 80% or more were determined to have a disinfecting effect.

[0070] [3. Evaluation method 2 for Acanthamoeba disinfection effect] Acanthamoeba concentration was 5 × 10 ６ An Acanthamoeba suspension was prepared and disinfected in the same manner as in [2. Method 1 for evaluating the disinfecting effect against Acanthamoeba] above, except that the concentration was adjusted to 100 cells / mL.

[0071] To 1 mL of the disinfected test solution, 9 mL of an inactivating agent (1 / 4 Ringer's solution containing 3.5% lecithin and 1.5% polysorbate) was added, and then a 10-fold serial dilution was prepared with 1 / 4 Ringer's solution. －１ ~10 －６ A diluted treatment solution of 100 ml was obtained.

[0072] 2 mL of 1.5% agar solution was added to a 12-well culture plate in advance, and E. coli (ATCC8739) suspension (approximately 1 × 10 １０ 0.1 mL of diluted treatment solution (10 －１ ~10 －６ ) was dispensed into 4 wells of the culture plate. The culture plate was cultured at 28°C for 14 days.

[0073] By culturing as described above, if Acanthamoeba survives, it will prey on E. coli and grow. On the other hand, if Acanthamoeba is dead, E. coli will grow. Using a control solution containing 1 / 4 Ringer's solution to which Acanthamoeba suspension has been added, the number of positive and negative wells in each dilution series was counted using the Reed and Muench method, and the LD was calculated from the total number of wells. ５０ The log reduction for each test solution was calculated by: Test solutions that were found to have a log reduction of 0.5 or more were determined to have a disinfecting effect.

[0074] 4. Method for Evaluating Antifungal Effect To evaluate the antifungal effect, Candida albicans (ATCC10231) was used as the test strain.

[0075] Candida was cultured on a slant of agar medium at 35°C for 18 hours, and then suspended in Dulbecco's phosphate buffer. The collected suspension was used as a bacterial stock solution. The bacterial stock solution was diluted with Dulbecco's phosphate buffer to a concentration of 10. ５ ~10 ６ The concentration was adjusted to cfu / mL to obtain a bacterial suspension.

[0076] 0.1 mL of the bacterial suspension was added to 10 mL of the test liquid preparation of each of the Examples and Comparative Examples for inoculation. After inoculation, the test liquid preparation was incubated at 25°C for 4 hours and 6 hours for disinfection.

[0077] One mL of the disinfected test solution was added to 9 mL of neutralizing broth containing an inactivator, and then serial 10-fold dilutions were prepared with Dulbecco's phosphate buffer to obtain the disinfected diluted test solution. One mL of each solution at three appropriate dilutions was dispensed into a disposable dish, and Sabouraud glucose agar medium was then poured into the dish and mixed. The dish was then incubated at 35°C for 48 hours, and the viable cell count was determined from the number of colonies obtained.

[0078] The initial bacterial count, which is the number of bacteria in the bacterial suspension, was determined by serially preparing a 10-fold dilution series with Dulbecco's phosphate buffer and then by the agar plate method as described above.

[0079] The log reduction after disinfection for each test liquid was calculated from the initial bacterial count and the viable bacterial count obtained using the disinfected diluted test liquid, using the following formula: Log reduction = initial bacterial count - viable bacterial count of test sample

[0080] The disinfection effect was judged according to the primary standard of the ISO stand-alone test, and test liquids with a logarithmic difference of 1.0 or more at a given disinfection time were judged to have a disinfection effect (reference: ISO 14729).

[0081] [5. Safety Evaluation Method] CO2 set at 37°C ２ Chinese hamster lung-derived V79 cells were pre-cultured in an incubator and then recovered by trypsinization, and then suspended in Eagle's minimum essential medium containing fetal bovine serum as a cell culture medium to obtain a cell suspension.

[0082] The test liquid preparations of the Examples and Comparative Examples were dispensed into the wells of a culture plate at liquid concentrations of 20%, 10%, 5%, and 2.5% (v / v%), followed by the addition of a cell suspension to give a cell count of 100, followed by the addition of Eagle's minimum essential medium containing fetal bovine serum to give a volume of 2,000 μL per well. For example, a 20% liquid preparation was prepared by mixing 1,500 μL of medium, 100 μL of cell suspension (1,000 cells / mL), and 400 μL of the test liquid preparation. The plate was then incubated in a CO 2 -storage system set at 37°C. ２ The cells were incubated in an incubator for 7 days. As a control, cells were cultured in cell culture medium alone without adding the test liquid. The relative colony formation rate at each concentration point of each test liquid was calculated using the following formula: Relative colony formation rate (%) = (number of colonies at each concentration point / number of colonies in the control) x 100

[0083] The relative colony formation rate was taken as the safety to ocular tissue, and the safety of each test solution was evaluated as follows by comparing the relative colony formation rates at the same concentration.

[0084] ++: Relative colony formation rate 75% or more ±: Relative colony formation rate over 50% and less than 75% -: Relative colony formation rate 50% or less

[0085] 6. Evaluation of Contact Lens Suitability The test lenses used were SEED 2weekFineUV Plus (Group 1), SEED 2weekPure (Group 4), J&J ACUVUE OASYS (Group VC), Alcon AIR OPTIX HydraGlyde (Group VC), and CooperVision Biofinity (Group VC).

[0086] Each test lens was removed from the blister pack, and water on the surface was removed. After water removal, the test lens was immersed in Dulbecco's phosphate buffer solution, and each parameter (power, base curve (BC), and diameter (DIA)) before treatment with the test solution was measured.

[0087] Next, each test lens was subjected to 30 hand rubs (cycle treatment) using the test liquid preparations of the Examples and Comparative Examples. After the cycle treatment, the test lens was immersed in a phosphate buffer solution, and the above parameters were measured.

[0088] The compatibility of each test solution with contact lenses was evaluated based on the amount of change before and after the cycle treatment in accordance with the contact lens approval standards shown below.

[0089] +: The fluctuation amount of Power is within ±0.25D, the fluctuation amount of BC is within ±0.2mm, and the fluctuation amount of DIA is within ±0.2mm. -: The fluctuation amount of Power exceeds ±0.25D, the fluctuation amount of BC exceeds ±0.2mm, or the fluctuation amount of DIA exceeds ±0.2mm.

[0090] [7. Method for Evaluating Physical Properties of Test Liquid Preparations] For the test liquid preparations of the Examples and Comparative Examples, the pH value was measured at room temperature using a pH meter (main body: Seven Compact S220, electrode: InLab MicroPro-ISM) (manufactured by Mettler Toledo), and the osmotic pressure was measured using an osmometer (manufactured by Advanced Instruments) "AD-3250."

[0091] [8. Evaluation Results] The measurement results of the pH value and osmotic pressure (mOsm) of the test solutions are shown in Tables 1A to 1C. The pH of the test solutions of Examples 1 to 22 was 6.55 to 8.52, and the osmotic pressure was 230 to 339. Therefore, it was found that these test solutions are suitable as solutions for contact lenses.

[0092] The test solutions of Examples 1 to 11 and Comparative Examples 1 to 6 were evaluated for their disinfecting effect against Acanthamoeba using Method 1 for Evaluating Acanthamoeba Disinfecting Effect. The results are shown in Tables 1A and 1C. As shown in the tables, the test solutions of Examples 1 to 11, which contained a combination of sodium dihydrogen phosphate dihydrate, a basic tonicity agent, and polydiallyldimethylammonium chloride having an average molecular weight of 10,000 to 1,100,000, all had a dead amoeba rate of 80% or higher, demonstrating sufficient disinfecting effect against Acanthamoeba. In contrast, the test solutions of Comparative Examples 1 to 6, which contained sodium dihydrogen phosphate dihydrate and a basic tonicity agent but either polydiallyldimethylammonium chloride having an average molecular weight of 8,500 or 1,200,000 or no polydiallyldimethylammonium chloride, all had a dead amoeba rate of 70% or lower, demonstrating no disinfecting effect against Acanthamoeba.

[0093] Furthermore, the test liquid preparations in Examples 1 to 11, which were found to have an Acanthamoeba disinfection effect, had a ratio of the content of the basic tonicity agent to the content of sodium dihydrogen phosphate ([basic tonicity agent] / [sodium dihydrogen phosphate]) within the range of 0.01 to 2, while the test liquid preparations in Comparative Examples 3 to 6, which were found not to have an Acanthamoeba disinfection effect, did not have this ratio within the above range.

[0094] The test solutions of Examples 12-22 and Comparative Examples 7-8 were evaluated for their disinfecting effect against Acanthamoeba using Method 2 for Evaluating Acanthamoeba Disinfecting Effect. The results are shown in Tables 1B and 1C. As shown in the tables, the test solutions of Examples 12-22, which contained a combination of sodium dihydrogen phosphate dihydrate, a basic tonicity agent, and polydiallyldimethylammonium chloride having an average molecular weight of 10,000 to 1,100,000, had logarithmic differences of 2.00 to 2.33, all of which were 0.5 or greater, demonstrating sufficient disinfecting effect against Acanthamoeba. In contrast, the test solutions of Comparative Examples 7-8, which contained sodium dihydrogen phosphate dihydrate and a basic tonicity agent but either contained polydiallyldimethylammonium chloride having an average molecular weight of 8,500 or did not contain polydiallyldimethylammonium chloride, had logarithmic differences of 0.25, demonstrating no disinfecting effect against Acanthamoeba.

[0095] Therefore, the evaluation method for the disinfection effect against Acanthamoeba conducted this time showed that a liquid containing a combination of sodium dihydrogen phosphate dihydrate, a basic isotonic agent, and polydiallyldimethylammonium chloride with an average molecular weight of 10,000 to 1,100,000 has an excellent disinfection effect against Acanthamoeba.

[0096] The test solutions of Examples 12-15 and 17-22 and Comparative Examples 7-8 were evaluated for their antifungal efficacy using the antifungal efficacy evaluation method. The results are shown in Tables 1B and 1C. As the tables show, even when the disinfection time was taken into account, the test solutions of the Examples had logarithmic differences of 1.0 or more and others less than 1.0, indicating that they did not necessarily have a disinfecting effect against fungi. In contrast, the test solution of Comparative Example 8, which contained polydiallyldimethylammonium chloride with an average molecular weight of 8,500, did not have a disinfecting effect against fungi. However, the test solution of Comparative Example 7, which contained sodium dihydrogen phosphate dihydrate and a basic tonicity agent but did not contain polydiallyldimethylammonium chloride, had a logarithmic difference of 1.0 or more at all disinfection times, demonstrating a disinfecting effect against fungi.

[0097] From the above results, it was surprisingly found that a liquid preparation containing polydiallyldimethylammonium chloride having an average molecular weight of 10,000 to 1,100,000 in addition to sodium dihydrogen phosphate dihydrate and a basic isotonic agent has a disinfecting effect specific to Acanthamoeba, although the disinfecting effect against fungi may be reduced in some cases compared to a liquid preparation not containing polydiallyldimethylammonium chloride.

[0098] When the test solutions of Examples 12 to 22 were evaluated for safety and contact lens compatibility, all test solutions were rated "+" at all concentrations evaluated, and the contact lens compatibility was rated "+" for all contact lenses tested. Taking the safety and contact lens compatibility evaluation results together with the pH and osmotic pressure evaluation results, it was found that solutions containing a combination of sodium dihydrogen phosphate dihydrate, a basic tonicity agent, and polydiallyldimethylammonium chloride having an average molecular weight of 10,000 to 1,100,000 have excellent disinfecting effects against Acanthamoeba and are suitable for contact lenses.

[0099]

[0100] The anti-Acanthamoeba liquid preparation of one embodiment of the present invention is highly safe for ocular tissues, has a wide range of application to contact lenses, and has excellent disinfecting effects against Acanthamoeba. Therefore, by using it as a contact lens care solution such as MPS, the risk of wearers contracting keratitis caused by Acanthamoeba can be reduced or avoided, thereby contributing to the health and welfare of contact lens wearers. CROSS-REFERENCE TO RELATED APPLICATIONS

[0101] This application claims priority from Japanese Patent Application No. 2023-123521, filed July 28, 2023, the entire disclosure of which is incorporated herein by reference. In addition, the entire disclosures of all documents referenced in the detailed description of the invention of this application, including Patent Documents 1 to 5, are incorporated herein by reference.

Claims

1. An anti-Acanthamoeba liquid preparation comprising sodium dihydrogen phosphate, a basic isotonic agent, and polydiallyldimethylammonium chloride having an average molecular weight of 10,000 to 1,100,000.

2. The anti-Acanthamoeba liquid preparation according to claim 1, wherein the ratio of the content of the basic isotonic agent to the content of the sodium dihydrogen phosphate ([basic isotonic agent] / [sodium dihydrogen phosphate]) is 0.01 to 2, and the pH is 6.5 to 8.

5.

3. The anti-Acanthamoeba liquid preparation according to claim 1, wherein the ratio of the content of the polydiallyldimethylammonium chloride to the content of the sodium dihydrogen phosphate ([polydiallyldimethylammonium chloride] / [sodium dihydrogen phosphate]) is 0.00005 to 0.

01.

4. The anti-Acanthamoeba liquid preparation according to claim 1, wherein the ratio of the content of the polydiallyldimethylammonium chloride to the total content of the sodium dihydrogen phosphate and the basic tonicity agent ([polydiallyldimethylammonium chloride] / ([sodium dihydrogen phosphate]+[basic tonicity agent]) is 0.000001 to 1.

5. The anti-Acanthamoeba liquid preparation according to claim 1, wherein the basic tonicity agent is one or more basic tonicity agents selected from the group consisting of 2-amino-2-hydroxymethyl-1,3-propanediol, 2-amino-2-ethyl-1,3-propanediol, arginine, and salts thereof.

6. The anti-Acanthamoeba liquid preparation according to claim 1, wherein the polydiallyldimethylammonium chloride has an average molecular weight of 100,000 to 1,000,000.

7. The anti-Acanthamoeba liquid preparation according to claim 1, further comprising an isotonicity agent other than the basic isotonicity agent.

8. The anti-Acanthamoeba liquid preparation according to claim 7, wherein the isotonicity agent is one or more isotonicity agents selected from the group consisting of sodium chloride, potassium chloride, 1,2-propanediol, 1,3-propanediol, mannitol and sorbitol.

9. The anti-Acanthamoeba liquid preparation according to claim 1, further comprising one or more quaternary ammonium salts selected from the group consisting of polyhexanide hydrochloride, alexidine, myristamidopropyl dimethylamine, benzalkonium chloride, benzethonium chloride and polidonium chloride.

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