Ophthalmic composition with reduced irritation

Abstract

The present invention addresses the problem of providing an ophthalmic composition wherein the sense of irritation imparted is suppressed, an irritation-inhibiting agent for an ophthalmic composition, a method for inhibiting irritation caused by an ophthalmic composition, and a method for assessing ophthalmic irritation. The problem is solved by an ophthalmic composition having inhibited irritating sensation and characterized by containing: one selected from the group consisting of (A-1) a hyperemia removal component, (A-2) an antihistamine component, (A-3) at least one selected from the group consisting of camphor, menthone, isomenthol, cineol, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) a local anesthetic component; and optionally, (B) a TRPA1 agonist.

Description

[Technical Field]

[0001] The present invention relates to an ophthalmic composition in which irritation is suppressed, an irritation inhibitor for the ophthalmic composition, a method for suppressing irritation from the ophthalmic composition, and a method for evaluating eye irritation. [Background technology]

[0002] Transient Receptor Potential (TRP) channels are cation channels with a transmembrane domain and are thought to function as tetramers. TRP channels are a family of cation channels that function in various aspects of sensory reception, and many TRP channel molecules exist that respond to diverse stimuli (Non-Patent Literature 1). TRPM8 is a cellular sensor involved in the sensation of coolness, TRPA1 in pain, and TRPV1 in burning. In the eye, TRPA1 and TRPV1 have also been reported to be involved in discomfort and pain (Non-Patent Literature 2).

[0003] l-Menthol is a TRPM8 agonist and is used as a cooling agent in eye drops and other medications, but it is also a TRPA1 agonist and is known to provide both a cooling and irritating sensation to the eyes. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Biochemistry, Vol. 81, No. 11, pp. 962-983, 2009. [Non-Patent Document 2] Curr. Ophthalmol. Rep. (2015), 3: 111-121 [Overview of the project] [Problems that the invention aims to solve]

[0005] The object of the present invention is to provide an ophthalmic composition in which irritation is suppressed, an irritation inhibitor for the ophthalmic composition, a method for suppressing irritation from the ophthalmic composition, and a method for evaluating eye irritation. [Means for solving the problem]

[0006] As a result of diligent research, the inventors have discovered that an ophthalmic composition containing a specific component can suppress the irritation to the eyes caused by TRPA1 agonists such as l-menthol, and in particular, can suppress the irritation while maintaining the cooling sensation of l-menthol, and furthermore, can solve the above-mentioned problems.

[0007] In other words, the present invention encompasses the following inventions. [1] An ophthalmic composition that suppresses irritation, characterized by containing at least one selected from the group consisting of the following components (A-1), (A-2), (A-3), and (A-4): (A-1) Decongestant component, (A-2) Antihistamine components, (A-3) At least one selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) Local anesthetic component. [2] An ophthalmic composition characterized by containing at least one selected from the group consisting of (A-1), (A-2), (A-3), and (A-4) below, and (B) a TRPA1 agonist, wherein irritation is suppressed: (A-1) Decongestant component, (A-2) Antihistamine components, (A-3) At least one selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) Local anesthetic component. [3] (B) The ophthalmic composition according to [2], wherein the TRPA1 agonist is l-menthol. [4] (A-1) The ophthalmic composition according to any one of [1] to [3] above, wherein the decongestant component is at least one selected from the group consisting of tetrahydrozoline, naphazoline, and pharmaceutically acceptable salts thereof. [5] (A-2) The ophthalmic composition according to any one of [1] to [3], wherein the antihistamine component is at least one selected from the group consisting of chlorpheniramine, diphenhydramine, and pharmaceutically acceptable salts thereof. [6] (A-4) The ophthalmic composition according to any one of [1] to [3] above, wherein the local anesthetic component is chlorobutanol. [7] An ophthalmic composition according to [1] to [6], wherein the irritation is pain caused by dry eye, allergies, etc. [8] An irritant inhibitor for ophthalmic compositions, characterized by containing at least one selected from the group consisting of the following components: (A-1), (A-2), (A-3), and (A-4): (A-1) Decongestant component, (A-2) Antihistamine components, (A-3) At least one selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) Local anesthetic component. [9] (B) An agent for suppressing irritation in an ophthalmic composition containing l-menthol, It is characterized by containing at least one selected from the group consisting of the following components (A-1), (A-2), (A-3), and (A-4). (B) Irritation inhibitors for ophthalmic compositions containing l-menthol: (A-1) Decongestant component, (A-2) Antihistamine components, (A-3) At least one selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) Local anesthetic component.

[10] An agent for suppressing pain caused by dry eyes, allergies, etc., characterized by containing at least one selected from the group consisting of the following components (A-1), (A-2), (A-3), and (A-4): (A-1) a decongestant component, (A-2) an antihistamine component, (A-3) at least one selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) a local anesthetic component.

[11] An ophthalmic composition for suppressing pain derived from dry eye, allergy or the like, characterized by comprising at least one selected from the group consisting of the following component (A-1), component (A-2), component (A-3), and component (A-4): (A-1) a decongestant component, (A-2) an antihistamine component, (A-3) at least one selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) a local anesthetic component.

[12] A method for suppressing irritation of an ophthalmic composition derived from (B) a TRPA1 agonist, characterized by further incorporating at least one selected from the group consisting of the following component (A-1), component (A-2), component (A-3), and component (A-4) into the ophthalmic composition containing (B) the TRPA1 agonist: (A-1) a decongestant component, (A-2) an antihistamine component, (A-3) at least one selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) a local anesthetic component.

[13] A method for evaluating eye irritation caused by a test sample, which comprises bringing the test sample into contact with TRPA1-expressing cells, and measuring the amount of change in intracellular calcium ion concentration induced via TRPA1 by the test sample.

[14] Use of at least one member selected from the group consisting of the following component (A-1), component (A-2), component (A-3), and component (A-4) for manufacturing an ophthalmic composition in which the irritating sensation of (B) TRPA1 agonist is suppressed: (A-1) Decongestant component, (A-2) Antihistamine component, (A-3) At least one member selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) Local anesthetic component.

[15] An agent for an ophthalmic composition containing (B) a TRPA1 agonist, for suppressing the irritating sensation while maintaining the refreshing sensation of the (B) TRPA1 agonist, which comprises at least one member selected from the group consisting of the following component (A-1), component (A-2), component (A-3), and component (A-4), and whereby the refreshing sensation of the (B) TRPA1 agonist is maintained at a level equivalent to that of the ophthalmic composition not containing component (A), while the irritating sensation is suppressed more than in the ophthalmic composition not containing component (A), Irritation suppressant for ophthalmic compositions containing (B) TRPA1 agonist: (A-1) Decongestant component, (A-2) Antihistamine component, (A-3) At least one member selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) Local anesthetic component.

[16] A method for suppressing the irritating sensation while maintaining the refreshing sensation provided by (B) a TRPA1 agonist, for an ophthalmic composition containing the (B) TRPA1 agonist, which comprises incorporating at least one member selected from the group consisting of the following component (A-1), component (A-2), component (A-3), and component (A-4), The ophthalmic composition is characterized in that the cooling sensation of the (B)TRPA1 agonist is maintained at the same level as the ophthalmic composition without component (A), while the irritation is suppressed compared to the ophthalmic composition without component (A). (B) Method for suppressing irritation in ophthalmic compositions containing a TRPA1 agonist: (A-1) Decongestant component, (A-2) Antihistamine components, (A-3) At least one selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) Local anesthetic component.

[17] At least one selected from the group consisting of the following components (A-1), (A-2), (A-3), and (A-4): (B) Use for the manufacture of ophthalmic compositions containing a TRPA1 agonist, The ophthalmic composition is characterized in that the cooling sensation of (B)TRPA1 agonist is maintained at the same level as that of an ophthalmic composition without component (A), while the irritation of (B)TRPA1 agonist is suppressed compared to an ophthalmic composition without component (A). Use: (A-1) Decongestant component, (A-2) Antihistamine components, (A-3) At least one selected from the group consisting of camphor, menthone, isomenthol, cineole, menthofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) Local anesthetic component. [Effects of the Invention]

[0008] The ophthalmic composition of the present invention is characterized by containing at least one selected from the group consisting of the following components (A-1), (A-2), (A-3), and (A-4), and further may or may not contain (B) a TRPA1 agonist, which has the effect of suppressing irritation: (A-1) a decongestant component, (A-2) an antihistamine component, (A-3) at least one selected from the group consisting of camphor, menthone, isomenthol, cineole, mentofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) a local anesthetic component. Preferably, the irritation is caused by a (B)TRPA1 agonist contained in the ophthalmic composition, but even when the ophthalmic composition does not contain a (B)TRPA1 agonist, it is preferable that the irritation is caused by pain symptoms in the animal, such as a human, being administered the ophthalmic composition, due to dry eye or allergies, or by activation of the TRPA1 irritation receptor in the animal. In particular, when the (B)TRPA1 agonist contained in the ophthalmic composition is menthol, it is possible to suppress irritation while maintaining a cooling sensation. Furthermore, the present invention also provides a method for suppressing irritation from ophthalmic compositions and a method for evaluating eye irritation caused by test samples. [Brief explanation of the drawing]

[0009] [Figure 1] Figure 1 shows the results of a test to confirm the suppression of irritation by the components constituting the ophthalmic composition of the present invention against the TRPA1 agonist (l-menthol). [Figure 2] Figure 2 visualizes the TRPA1 inhibitory effect, showing the results for borneol (reference example), tetrahydrozoline hydrochloride, and peppermint oil (example). [Figure 3] Figure 3 shows the results of a human cooling and stimulating sensation test using l-menthol (comparative example) and a combination of l-menthol and tetrahydrozoline hydrochloride (example). [Modes for carrying out the invention]

[0010] The ophthalmic composition of the present invention comprises at least one selected from the group consisting of the following components: (A-1) (A-2) (A-3) and (A-4): (A-1) an anticongestive component, (A-2) an antihistamine component, (A-3) at least one selected from the group consisting of camphor, menthone, isomenthol, cineole, mentofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) a local anesthetic component. Preferably, it contains at least one selected from the group consisting of the group consisting of the components (A-1) (A-2) (A-3) and (A-4) as an active ingredient. In this specification, unless otherwise specified, the unit of content "%" means "w / v%" and is synonymous with "g / 100mL".

[0011] [(A-1) Decongestant component] As the decongestant component, it is preferable to use at least one selected from the group consisting of imidazoline-based decongestant components such as tetrahydrozoline, naphazoline, oxymetazoline, or their hydrochloride salts and nitrates, epinephrine, ephedrine, methylephedrine, phenylephrine, and pharmaceutically acceptable salts thereof, but is not limited to these. Of these decongestant components, preferably, for example, imidazoline-based decongestant components, more preferably tetrahydrozoline, naphazoline and pharmaceutically acceptable salts thereof, even more preferably tetrahydrozoline and pharmaceutically acceptable salts thereof, and particularly preferably tetrahydrozoline hydrochloride (tetrahydrozoline hydrochloride).

[0012] When the ophthalmic composition of the present invention contains component (A-1), the content of component (A-1) is not particularly limited and is set appropriately according to the type of component (A-1), the types and content of other components, the use of the ophthalmic composition, and the formulation form. From the viewpoint of more significantly demonstrating the effects of the present invention, the lower limit of the total content of component (A-1) relative to the total amount of the ophthalmic composition is preferably 0.0001 w / v% or more, 0.0003 w / v% or more, or 0.0005 w / v% or more. Furthermore, 0.01 w / v% or more is even more preferable. Also, when the ophthalmic composition of the present invention contains component (A-1), from the viewpoint of more significantly demonstrating the effects of the present invention, the upper limit of the content of component (A-1) relative to the total amount of the ophthalmic composition is preferably 0.5 w / v% or less, 0.3 w / v% or less, 0.1 w / v% or less, or 0.05 w / v% or less. Furthermore, the lower and upper limits mentioned above may be combined in any way.

[0013] When the ophthalmic composition of the present invention contains component (A-1), the content ratio of component (A-1) to component (B) is not particularly limited and is set appropriately depending on the type of component (A-1), the types and content of other components, the use of the ophthalmic composition and the formulation form. As for the content ratio of component (A-1) to component (B), from the viewpoint of more significantly exhibiting the effects of the present invention, for example, the total content of component (A-1) contained in the ophthalmic composition of the present invention is, for example, usually 0.001 parts by weight or more, preferably 0.01 parts by weight or more, more preferably 0.1 parts by weight or more, even more preferably 0.2 parts by weight or more, and particularly preferably 0.5 parts by weight or more, per 1 part by weight of the total content of component (B). Furthermore, the lower and upper limits mentioned above may be combined in any way.

[0014] [(A-2) Antihistamine components] As the antihistamine component, it is preferable to use at least one selected from the group consisting of, for example, chlorpheniramine, diphenhydramine, ketotifen, iproheptine, levocabastine, olopatadine, and pharmaceutically acceptable salts thereof, but it is not limited to these. Among these antihistamine components, preferred are ketotifen fumarate, iproheptin, diphenhydramine hydrochloride, chlorpheniramine maleate (chlorpheniramine maleate), levocabastine hydrochloride, olopatadine hydrochloride, and the like. More preferably, ketotifen fumarate, diphenhydramine hydrochloride, chlorpheniramine maleate, levocabastine hydrochloride, olopatadine hydrochloride, and the like. Even more preferably, diphenhydramine hydrochloride and chlorpheniramine maleate are used, and even more preferably, chlorpheniramine maleate is used.

[0015] When the ophthalmic composition of the present invention contains component (A-2), the content of component (A-2) is not particularly limited and is set appropriately according to the type of component (A-2), the types and content of other components, the use of the ophthalmic composition, and the formulation form. From the viewpoint of more significantly demonstrating the effects of the present invention, it is preferable that the lower limit of the total content of component (A-2) relative to the total amount of the ophthalmic composition is, for example, 0.001 w / v% or more, 0.003 w / v% or more, 0.005 w / v% or more, or 0.01 w / v% or more. Furthermore, when the ophthalmic composition of the present invention contains component (A-2), from the viewpoint of more significantly demonstrating the effects of the present invention, it is preferable that the upper limit of the total content of component (A-2) relative to the total amount of the ophthalmic composition is, for example, 0.5 w / v% or less, 0.3 w / v% or less, 0.05 w / v% or less, or 0.03 w / v% or less. Furthermore, the lower and upper limits mentioned above may be combined in any way.

[0016] When the ophthalmic composition of the present invention contains component (A-2), the content ratio of component (A-2) to component (B) is not particularly limited and is set appropriately depending on the type of component (A-2), the types and content of other components, the use of the ophthalmic composition and the formulation form. As for the content ratio of component (A-2) to component (B), from the viewpoint of more significantly exhibiting the effects of the present invention, for example, the total content of component (A-2) contained in the ophthalmic composition of the present invention is preferably, for example, at least 0.002 parts by weight, more preferably 0.004 parts by weight, even more preferably 0.01 parts by weight, even more preferably 0.05 parts by weight, and particularly preferably 0.1 parts by weight or more, per 1 part by weight of the total content of component (B). Furthermore, the proportion of component (A-2) is preferably, for example, 50,000 parts by weight or less, more preferably 5,000 parts by weight or less, even more preferably 1,000 parts by weight or less, even more preferably 800 parts by weight or less, even more preferably 500 parts by weight or less, even more preferably 100 parts by weight or less, and particularly preferably 10 parts by weight or less, relative to 1 part by weight of component (B). Furthermore, the lower and upper limits mentioned above may be combined in any way.

[0017] [Component (A-3)] The ophthalmic composition of the present invention contains, as component (A-3), at least one selected from the group consisting of camphor, menthone, isomenthol, cineole, mentofuran, pinene, limonene, neomenthol, and methyl acetate. These may be d-isomers, l-isomers, or dl-isomers.

[0018] Furthermore, component (A-3) of the present invention can be used as an essential oil containing component (A-3). Examples of essential oils containing component (A-3) include one or more of the following: peppermint oil, cool mint oil, spearmint oil, fennel oil, cinnamon oil, bergamot oil, eucalyptus oil, cinnamon oil, and rose oil, with peppermint oil or cool mint oil being particularly preferred.

[0019] If the ophthalmic composition of the present invention contains component (A-3), the proportion of component (A-3) may be, as the total amount of component (A-3) relative to the total amount of the ophthalmic composition, for example, with a lower limit of 0.00001 w / v% or more, 0.00005 w / v% or more, 0.0001 w / v% or more, 0.0003 w / v% or more, 0.0005 w / v% or more, 0.0007 w / v% or more, 0.0008 w / v% or more, 0.0009 w / v% or more, or 0.001 w / v% or more. Furthermore, if the ophthalmic composition of the present invention contains component (A-3), the proportion of component (A-3) may be, for example, up to 5 w / v% or less, 3 w / v% or less, 2 w / v% or less, 1.5 w / v% or less, 1.2 w / v% or less, 1 w / v% or less, 0.9 w / v% or less, 0.8 w / v% or less, 0.7 w / v% or less, 0.6 w / v% or less, 0.5 w / v% or less, 0.3 w / v% or less, or 0.1 w / v% or less, relative to the total amount of the ophthalmic composition. Furthermore, the lower and upper limits mentioned above may be combined in any way. In particular, the proportion of component (A-3) may be, for example, 0.00001 w / v% to 2 w / v% (for example, 0.00005 to 1 w / v%) in total with respect to the total amount of the ophthalmic composition, preferably 0.0001 to 0.1 w / v% (for example, 0.0003 to 0.05 w / v%) in total, and more preferably 0.0005 to 0.02 w / v% (for example, 0.001 to 0.01 w / v%) in total. Furthermore, the proportion of component (A-3) may be, for example, lower limits of 0.00000005% or more, 0.0000005% or more, or 0.000001% or more with respect to the total amount of the ophthalmic composition. (A-3) The origin of the components is not particularly limited and may include those derived from essential oils.

[0020] When the ophthalmic composition of the present invention contains component (A-3), the content ratio of component (A-3) to component (B) is not particularly limited and is set appropriately depending on the type of component (A-3), the types and content of other components, the use of the ophthalmic composition and the formulation form. As for the content ratio of component (A-3) to component (B), from the viewpoint of more significantly exhibiting the effects of the present invention, the lower limit may be, for example, 0.000001 parts by weight or more, 0.00001 parts by weight or more, 0.0001 parts by weight or more, 0.001 parts by weight or more, 0.005 parts by weight or more, 0.01 parts by weight or more, 0.1, or 1 part by weight or more per 1 part by weight of the total content of component (B). Furthermore, the proportion of component (A-3) may be, for example, up to 500,000 parts by weight, 300,000 parts by weight or less, 15,000 parts by weight or less, 800 parts by weight or less, 500 parts by weight or less, 100 parts by weight or less, or 10 parts by weight or less, relative to 1 part by weight of component (B). Furthermore, the lower and upper limits mentioned above may be combined in any way.

[0021] [Component (A-4)] Chlorobutanol is preferred as the local anesthetic component. In addition, in the present invention, local anesthetic components known in the art, such as procaine hydrochloride or lidocaine hydrochloride, may be used instead of chlorobutanol, or chlorobutanol may be used in combination with other known local anesthetic components.

[0022] In particular, when the ophthalmic composition of the present invention contains component (A-4), the proportion is preferably, for example, 0.001 to 2.5 w / v%, more preferably 0.01 to 1 w / v%, even more preferably 0.05 to 0.5 w / v%, and particularly preferably 0.1 to 0.25% relative to the total amount of the ophthalmic composition.

[0023] When the ophthalmic composition of the present invention contains component (A-4), the content ratio of component (A-4) to component (B) is not particularly limited and is set appropriately depending on the type of component (A-4), the types and content of other components, the use of the ophthalmic composition, and the formulation form. As for the content ratio of component (A-4) to component (B), from the viewpoint of more significantly exhibiting the effects of the present invention, the lower limit may be, for example, 0.01 parts by weight or more, 0.1 parts by weight or more, or 1 part by weight or more, per 1 part by weight of the total content of component (B). Also, the proportion of component (A-3) may have an upper limit of, for example, 5000 parts by weight or less, 3500 parts by weight or less, 1000 parts by weight or less, 500 parts by weight or less, 100 parts by weight or less, 50 parts by weight or less, or 10 parts by weight or less, per 1 part by weight of component (B). Furthermore, the lower and upper limits mentioned above may be combined in any way.

[0024] [Component (A)] In this specification, components (A-1), (A-2), (A-3), and (A-4) may be collectively referred to as component (A). Furthermore, it is preferable to use multiple components in combination, more preferably to use two or more groups selected from the group consisting of (A-1) to (A-4), and even more preferably to use three or more groups in combination.

[0025] [(B) Component: TRPA1 agonist] The ophthalmic composition of the present invention preferably contains a TRPA1 agonist. The TRPA1 agonist provides a stimulating sensation to the eye. Menthol is preferred as the TRPA1 agonist. Menthol is not particularly limited as long as it is medically, pharmacologically, or physiologically acceptable, and may be in the d-isomer, l-isomer, or dl-isomer, but l-menthol is preferred.

[0026] Alternatively, essential oils containing menthol may be used as menthol.

[0027] The menthol content in the ophthalmic composition of the present invention is preferably 0.00001 to 0.5 w / v%, more preferably 0.0001 to 0.25 w / v%, even more preferably 0.0005 to 0.1 w / v%, and still more preferably 0.001 to 0.08 w / v% relative to the total amount of the ophthalmic composition. When using essential oils containing menthol, the total amount of menthol, including the amount of menthol contained in the essential oil, is set to satisfy the above proportions.

[0028] The ophthalmic composition of the present invention can suppress irritation by containing at least one component selected from the group consisting of (A-1), (A-2), (A-3), and (A-4). Furthermore, it is particularly preferable that the ophthalmic composition of the present invention contains (B) a TRPA1 agonist, as this can suppress irritation caused by the TRPA1 agonist contained in the ophthalmic composition. It is also preferable that the animal, such as a human, to be administered the ophthalmic composition exhibits pain symptoms due to dry eye or allergies, or that the TRPA1 irritant receptor in the animal is activated.

[0029] The ophthalmic composition of the present invention may further contain a nonionic surfactant in addition to the components described above. The nonionic surfactant is not particularly limited as long as it is pharmaceutically, pharmacologically, or physiologically acceptable. Specific examples of nonionic surfactants include, for example, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene hydrogenated castor oil, polyoxyethylene castor oil, polyoxyethylene-polyoxypropylene block copolymer, polyoxyethylene-polyoxypropylene block copolymer adducts, polyoxyethylene-polyoxypropylene alkyl ethers, and polyoxyethylene alkylphenyl ethers.

[0030] More specific examples of nonionic surfactants include, for example, polyoxyethylene (hereinafter also referred to as POE)-polyoxypropylene (hereinafter also referred to as POP) glycol (e.g., poloxamers such as poloxamer 407, poloxamer 235, poloxamer 188); POE-POP block copolymer adducts of ethylenediamines such as poloxamine; POE(20) sorbitan monolauric acid (polysorbate 20), POE(20) sorbitan monooleate (polysorbate 80), POE sorbitan monostearate (polysorbate 60), POE sorbitan tristearate (polysorbate 65), and other POE(5) hard Examples include hydrogenated castor oils such as POE(10) hydrogenated castor oil, POE(20) hydrogenated castor oil, POE(40) hydrogenated castor oil, POE(50) hydrogenated castor oil, POE(60) hydrogenated castor oil, and POE(100) hydrogenated castor oil; POE castor oils such as POE(3) castor oil, POE(10) castor oil, and POE(35) castor oil; POE alkyl ethers such as POE(9) lauryl ether; POE·POP alkyl ethers such as POE(20)POP(4) cetyl ether; POE alkylphenyl ethers such as POE(10) nonylphenyl ether; and polyethylene glycol monostearate such as polyoxyl 40 stearate. The numbers in parentheses indicate the average number of moles of POP or POE added.

[0031] Among these nonionic surfactants, POE-POP glycol, POE sorbitan fatty acid esters, POE hydrogenated castor oil, POE castor oil, and polyethylene glycol monostearate are preferred from the viewpoint of significantly exhibiting the effects of the present invention, poloxamer 407, POE(20) sorbitan monooleate (polysorbate 80), POE(60) hydrogenated castor oil (polyoxyethylene hydrogenated castor oil 60), POE(40) hydrogenated castor oil (polyoxyethylene hydrogenated castor oil 40), POE(3) castor oil (polyoxyethylene castor oil 3), POE(10) castor oil (polyoxyethylene castor oil 10), POE(35) castor oil (polyoxyethylene castor oil 35), and polyoxyl stearate 40 are more preferred, and POE(20) sorbitan monooleate and POE(60) hydrogenated castor oil are particularly preferred.

[0032] These nonionic surfactants may be synthesized and used by known methods, or they may be obtained commercially and used. Nonionic surfactants may be used individually, or two or more may be used in any combination. The content of nonionic surfactants can be appropriately determined depending on the type of nonionic surfactant, the type and amount of other components, and the intended use of the ophthalmic composition.

[0033] Furthermore, the ophthalmic composition of the present invention may contain surfactants other than nonionic surfactants. Such surfactants are not particularly limited as long as they are pharmaceutically, pharmacologically, or physiologically acceptable, but specific examples include anionic surfactants such as polyoxyethylene alkyl ether phosphates, polyoxyethylene alkyl ether sulfates, alkylbenzene sulfonates, alkyl sulfates, and N-acyl taurine salts, or amphoteric surfactants such as lauryldimethylaminoacetic acid betaine.

[0034] As an example, the total amount of surfactant relative to the total amount of the ophthalmic composition is not limited, but for example, the lower limit is preferably 0.0001 w / v% or more, more preferably 0.0005 w / v% or more, even more preferably 0.001 w / v% or more, particularly preferably 0.005 w / v% or more, and most preferably 0.01 w / v% or more. The upper limit is not limited, but is preferably 10 w / v% or less, more preferably 5 w / v% or less, even more preferably 1 w / v% or less, and particularly preferably 0.5 w / v% or less. Furthermore, the above lower and upper limits may be combined in any way.

[0035] In the present invention, the ophthalmic composition may further contain a thickening agent in addition to the above-mentioned components. Specific examples of thickening agents include cellulosic polymer compounds such as methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose (hypromellose), carboxymethylcellulose and its salts, carboxyethylcellulose and its salts, glycosaminoglycans such as chondroitin sulfate and its salts, and hyaluronic acid and its salts, vinyl polymers such as polyvinyl alcohol (complete or partially saponified), polyvinylpyrrolidone (K17, K25, K30, K90, etc.), carboxyvinyl polymer and its salts, dextran, macrogol (polyethylene glycol) such as macrogol 6000, macrogol 4000, and macrogol 400, gellan gum, alginic acid and its salts. Here, the salts are not particularly limited, but sodium salts are preferred. From the viewpoint of significantly exhibiting the effects of the present invention, glycosaminoglycans, cellulosine polymers, or vinyl polymers are preferred, glycosaminoglycans are more preferred, chondroitin sulfate, hyaluronic acid, and their salts are even more preferred, and sodium chondroitin sulfate (sodium chondroitin sulfate ester) and sodium hyaluronate are even more preferred. These thickeners may be synthesized and used by known methods, or commercially available products may be used. These thickeners may be used individually or in combination of two or more.

[0036] The total content of the thickening agent can be appropriately set depending on the type of thickening agent used, the type and amount of other ingredients, and the intended use of the ophthalmic composition. The total content of the thickening agent relative to the total amount of the ophthalmic composition is not limited, but is preferably 0.0001 w / v% or more, more preferably 0.0005 w / v% or more, even more preferably 0.001 w / v% or more, even more preferably 0.005 w / v% or more, even more preferably 0.01 w / v% or more, and most preferably 0.05 w / v% or more. Furthermore, the total content of the thickening agent relative to the total amount of the ophthalmic composition is not limited, but is preferably 5 w / v% or less, more preferably 3 w / v% or less, even more preferably 1 w / v% or less, even more preferably 0.5 w / v% or less, and most preferably 0.3 w / v% or less. The total amount of the thickening agent relative to the total amount of the ophthalmic composition is not limited, but is preferably 0.0001 to 5 w / v%, more preferably 0.0005 to 3 w / v%, even more preferably 0.001 to 1 w / v%, even more preferably 0.005 to 0.5 w / v%, particularly preferably 0.01 to 0.5 w / v%, and most preferably 0.05 to 0.3 w / v%.

[0037] In the present invention, the ophthalmic composition may further contain a buffering agent in addition to the components described above. The buffering agent is not particularly limited as long as it is pharmaceutically, pharmacologically, or physiologically acceptable. Examples of such buffering agents include borate buffers, phosphate buffers, carbonate buffers, citrate buffers, acetate buffers, Tris buffers, aspartic acid, aspartate salts, epsilon-aminocaproic acid, etc. Borate buffers, phosphate buffers, carbonate buffers, and citrate buffers are preferred as buffering agents.

[0038] As more specific examples, though not limited to them, examples of boric acid buffers include boric acid or its salts (sodium borate, potassium tetraborate, potassium metaborate, ammonium borate, borax, etc.); phosphoric acid or its salts (disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, trisodium phosphate, dipotassium phosphate, calcium monohydrogen phosphate, calcium dihydrogen phosphate, etc.) as phosphate buffers; carbonate or its salts (sodium bicarbonate, sodium carbonate, ammonium carbonate, potassium carbonate, calcium carbonate, potassium bicarbonate, magnesium carbonate, etc.) as carbonate buffers; citric acid or its salts (sodium citrate, potassium citrate, calcium citrate, sodium dihydrogen citrate, disodium citrate, etc.) as citric acid buffers; acetic acid or its salts (ammonium acetate, potassium acetate, calcium acetate, sodium acetate, etc.) as acetic acid buffers; and aspartic acid or its salts (sodium aspartate, magnesium aspartate, potassium aspartate, magnesium-potassium aspartate mixture, etc.). Furthermore, hydrates of each salt may be used as raw materials for the manufacture of buffering agents.

[0039] These buffering materials may be synthesized using known methods or obtained commercially and used. These buffering materials may be used individually or in any combination of two or more materials.

[0040] The buffering agent content in the ophthalmic composition of the present invention can be appropriately set depending on the type of buffering agent used, the type and amount of other components, the intended use of the ophthalmic composition, etc. As an example, the total buffering agent content relative to the total amount of the ophthalmic composition is not limited, but is preferably 0.01 w / v% or more, more preferably 0.05 w / v% or more, even more preferably 0.1 w / v% or more, and particularly preferably 0.3 w / v% or more. The total buffering agent content relative to the total amount of the ophthalmic composition is not limited, but is preferably 10 w / v% or less, more preferably 5 w / v% or less, even more preferably 3 w / v% or less, and particularly preferably 2 w / v% or less. The total buffering agent content relative to the total amount of the ophthalmic composition is not limited, but is preferably 0.01 to 10 w / v%, more preferably 0.05 to 5 w / v%, even more preferably 0.1 to 3 w / v%, and particularly preferably 0.3 to 2 w / v%.

[0041] In the present invention, the ophthalmic composition may further contain an isotonic agent in addition to the components described above. The isotonic agent is not particularly limited as long as it is pharmaceutically, pharmacologically, or physiologically acceptable. Examples of isotonic agents include sodium bisulfite, sodium sulfite, potassium chloride, calcium chloride, sodium chloride, magnesium chloride, sodium thiosulfate, magnesium sulfate, glycerin, and propylene glycol. Among these isotonic agents, sodium chloride, potassium chloride, calcium chloride, magnesium chloride, glycerin, and propylene glycol are preferred from the viewpoint of significantly exhibiting the effects of the present invention. These isotonic agents may be used individually or in any combination of two or more.

[0042] The content of the isotonic agent in the ophthalmic composition of the present invention can be appropriately set depending on the type of isotonic agent used, the type and amount of other components, and the intended use of the ophthalmic composition. As an example, the total content of the buffering agent relative to the total amount of the ophthalmic composition is not limited, but is preferably 0.01 to 10 w / v%, preferably 0.05 to 5 w / v%, and more preferably 0.1 to 3 w / v%.

[0043] The ophthalmic composition of the present invention may contain, in accordance with conventional methods, various additives selected appropriately and used in combination in appropriate amounts, one or more types, depending on its intended use and formulation, as long as the effects of the present invention are not impaired. Examples of such additives include the various additives listed as active ingredients (pharmacologically active ingredients, physiologically active ingredients, etc.) in the 2016 Dictionary of Pharmaceutical Additives (edited by the Japan Pharmaceutical Additives Association) or the 2012 Edition of the Standards for Approval of Manufacturing and Marketing of Over-the-Counter Drugs (Japan Regulatory Science Society). Representative additives include the following: Carrier: For example, an aqueous solvent such as water or aqueous ethanol. Chelating agents: For example, ethylenediaminediacetic acid (EDDA), ethylenediaminetriacetic acid, ethylenediaminetetraacetic acid (EDTA), N-(2-hydroxyethyl)ethylenediaminetriacetic acid (HEDTA), diethylenetriaminepentaacetic acid (DTPA), etc. Base: For example, octyldodecanol, titanium dioxide, potassium bromide, Plastibase, etc.

[0044] pH adjusters: For example, hydrochloric acid, acetic acid, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, triethanolamine, diisopropanolamine, etc. Stabilizers: For example, sodium formaldehyde sulfoxylate (Longalit), sodium bisulfite, sodium pyrosulfite, aluminum monostearate, glyceryl monostearate, cyclodextrin, monoethanolamine, etc. Preservatives, disinfectants, or antibacterial agents: for example, zinc chloride, alkyldiaminoethylglycine hydrochloride, sodium benzoate, ethanol, benzalkonium chloride, benzethonium chloride, chlorhexidine gluconate, sorbic acid, potassium sorbate, sodium dehydroacetate, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, butyl parahydroxybenzoate, oxyquinoline sulfate, phenethyl alcohol, benzyl alcohol, biguanide compounds (specifically, polyhexanide hydrochloride, etc.), Glokill (a trade name of Rhodia Corporation), etc.

[0045] Sugars: For example, glucose, cyclodextrin, alpha-cyclodextrin, etc. Sugar alcohols: For example, xylitol, sorbitol, mannitol, glycerin, etc. These may be in d-isomer, l-isomer, or dl-isomer.

[0046] Furthermore, the ophthalmic composition with suppressed irritation according to this embodiment may contain various pharmacologically active and physiologically active components in appropriate amounts in combination with the above components, as long as they do not interfere with the effects of the present invention. The components are not particularly limited, and examples include active ingredients in various pharmaceuticals listed in the 2012 edition of the Standards for Approval of Manufacturing and Marketing of Over-the-Counter Drugs (supervised by the Regulatory Science Society of Japan). For example, the following components can be specifically used in ophthalmic drugs. Ocular muscle modulating agents: For example, neostigmine methylsulfate, tropicamide, cholinesterase inhibitors such as helenien sulfate atropine, etc. Vitamins: For example, flavin adenine dinucleotide (FAD), flavin adenine dinucleotide sodium, pyridoxine hydrochloride, cyanocobalamin, retinyl acetate, retinyl palmitate, panthenol, ascorbic acid, tocopherol acetate, d-α-tocopherol acetate, tocopherol succinate, tocopherol nicotinate, tocopherol linolenic acid, etc. Amino acids: For example, L-glutamic acid, potassium L-aspartate, magnesium potassium L-aspartate, aminoethylsulfonic acid (taurine), etc. Anti-inflammatory agents: For example, glycyrrhizic acid, dipotassium glycyrrhizinate, glycyrrhetin, methyl salicylate, glycol salicylate, azulene sulfonic acid, sodium azulene sulfonate, allantoin, tranexamic acid, berberine, lysozyme, lysozyme chloride, berberine chloride, berberine sulfate, berberine sulfate hydrate, epsilon-aminocaproic acid, indomethacin, pranoprofen, ibuprofen, ibuprofen piconol, ketoprofen, diclofenac sodium, bromfenac sodium, felbinac, bendazac, piroxicam, bufexamac, butyl flufenamate, zinc sulfate hydrate, etc. Astringent agents: For example, zinc oxide, zinc lactate, zinc sulfate, etc. These may also be in hydrate form. Switching components: For example, sodium cromoglycate, pranoprofen, etc.

[0047] Others: For example, sulfamethoxazole, sulfamethoxazole sodium, sulfisoxazole, sulfisomidine sodium, gromwell root, horse chestnut, and their salts, creatinine, tyroxapole, sesame oil, castor oil, dibutylhydroxytoluene, phenylethyl alcohol, etc.

[0048] If the ophthalmic composition of the present invention contains water, from the viewpoint of more significantly exhibiting the effects of the present invention, for example, the water content based on the total amount of the ophthalmic composition is preferably 80 w / v% or more and less than 100 w / v%, more preferably 85 w / v% or more and 99.5 w / v%, and even more preferably 90 w / v% or more and 99.2 w / v%.

[0049] The water used in the ophthalmic composition of the present invention may be any water that is pharmaceutically, pharmacologically, or physiologically acceptable. Examples of such water include distilled water, tap water, purified water, sterile purified water, water for injection, and distilled water for injection. Their definitions are based on the 17th edition of the Japanese Pharmacopoeia.

[0050] The ophthalmic composition of the present invention can be prepared by adding and mixing one or more components selected from the group consisting of (A-1), (A-2), (A-3), and (A-4) in desired amounts, and optionally (B) a TRPA agonist, and other components as needed, to the desired concentration. For example, it can be prepared by dissolving or dispersing these components in purified water, adjusting to a predetermined pH and osmotic pressure, and sterilizing by filtration sterilization or the like. It is preferable that the ophthalmic composition of the present invention does not contain benzalkonium chloride or its salts, caffeine or its salts, or 50,000 units or more of retinol palmitate, but it may contain these.

[0051] The pH of the ophthalmic composition of the present invention is not particularly limited, as long as it is within a range that is pharmaceutically, pharmacologically, or physiologically acceptable. For example, the pH of the ophthalmic composition can be appropriately set depending on the use of the ophthalmic composition, the formulation form, the method of use, etc., but for example it may be 4.0 to 9.5, preferably 4.0 to 9.0, more preferably 4.5 to 9.0, even more preferably 4.5 to 8.5, and still more preferably 5.0 to 8.5.

[0052] The ophthalmic composition of the present invention can be adjusted to an osmotic pressure ratio within a range acceptable to the body, as needed. The appropriate osmotic pressure ratio can be set appropriately depending on the use, formulation form, and method of use of the ophthalmic composition, but for example, it can be 0.4 to 5.0, preferably 0.6 to 3.0, more preferably 0.8 to 2.2, and even more preferably 0.8 to 2.0. The osmotic pressure ratio is the ratio of the osmotic pressure of the sample to 286 mOsm (osmotic pressure of 0.9 w / v% sodium chloride aqueous solution) based on the 17th edition of the Japanese Pharmacopoeia, and the osmotic pressure is measured by referring to the osmotic pressure measurement method (freezing point depression method) described in the Japanese Pharmacopoeia. The standard solution for osmotic pressure ratio measurement (0.9 w / v% sodium chloride aqueous solution) can be prepared by drying sodium chloride (Japanese Pharmacopoeia standard reagent) at 500-650°C for 40-50 minutes, then allowing it to cool in a desiccator (silica gel), accurately weighing 0.900 g of it, dissolving it in purified water to make exactly 100 mL, or by using a commercially available standard solution for osmotic pressure ratio measurement (0.9 w / v% sodium chloride aqueous solution).

[0053] The viscosity of the ophthalmic composition of the present invention is not particularly limited, as long as it is within a range that is pharmaceutically, pharmacologically, or physiologically acceptable. For example, the viscosity at 20°C measured with a rotational viscometer (RE550 viscometer, manufactured by Toki Sangyo Co., Ltd., rotor: 1°34'×R24) is preferably 0.1 to 10000 mPa·s, more preferably 0.1 to 8000 mPa·s, even more preferably 0.1 to 1000 mPa·s, even more preferably 0.1 to 100 mPa·s, and particularly preferably 1 to 20 mPa·s.

[0054] The ophthalmic composition of the present invention can take various formulation forms depending on the purpose. Examples of such formulation forms include liquids, gels, and semi-solid preparations (such as ointments).

[0055] The ophthalmic composition of the present invention can be used, for example, as eye drops (also called eye solutions or eye medicines; eye drops include eye drops that can be used while wearing contact lenses), artificial tears, eye washes (also called eye wash solutions or eye medicines; eye washes include eye washes that can be used while wearing contact lenses), contact lens compositions [contact lens insertion solutions, contact lens care compositions (contact lens disinfectants, contact lens preservatives, contact lens cleaning agents, contact lens cleaning and preservation agents), contact lens packaging solutions, etc.]. Furthermore, "contact lenses" include hard contact lenses and soft contact lenses (which encompass both ionic and nonionic types, and both silicone hydrogel contact lenses and non-silicone hydrogel contact lenses).

[0056] The ophthalmic composition of the present invention is preferably an eye drop (including eye drops that can be instilled while wearing contact lenses) because it can more clearly demonstrate the effects of the present invention. When the ophthalmic composition according to this embodiment is an eye drop, the dosage and method of use are not particularly limited as long as they are effective and have few side effects, but for example, for adults (15 years of age or older) and children 7 years of age or older, 1 to 7 drops at a time is preferred, and 1 to 3 drops, 1 to 2 drops, or 2 to 3 drops at a time is more preferred. As for the number of times to instill, if 1 to 3 drops, 1 to 2 drops, or 2 to 3 drops are used at a time, an example of use is to instill 5 to 6 times a day. Alternatively, 4 to 6 drops may be instilled 3 to 6 times a day.

[0057] The ophthalmic composition of the present invention is provided in any container. The container for containing the ophthalmic composition according to this embodiment is not particularly limited and may be made of glass or plastic, for example. Plastic is preferred. Examples of plastics include polyethylene terephthalate, polyarylate, polyethylene naphthalate, polycarbonate, polyethylene, polypropylene, polyimide and copolymers of monomers constituting these, and mixtures of two or more of these. Polyethylene terephthalate is preferred. The container for containing the ophthalmic composition according to this embodiment may be a transparent container that allows visibility of the contents inside, or an opaque container that makes it difficult to see the contents inside. A transparent container is preferred. Here, "transparent container" includes both colorless transparent containers and colored transparent containers.

[0058] A nozzle may be attached to the container for containing the ophthalmic composition of the present invention. The material of the nozzle is not particularly limited; for example, it may be made of glass or plastic. Plastic is preferred. Examples of plastics include: Examples include polybutylene terephthalate, polyethylene, polypropylene, polyethylene terephthalate, polyethylene naphthalate, copolymers of monomers comprising these materials, and mixtures of two or more of these materials.

[0059] The container for containing the ophthalmic composition of the present invention may be a multi-dose type that contains multiple doses, or a unit-dose type that contains a single dose.

[0060] [Ophthalmic compositions or pain relievers for the suppression of pain caused by dry eyes, allergies, etc.] Furthermore, by containing at least one selected from the group consisting of components (A-1), (A-2), (A-3), and (A-4) above, the ophthalmic composition of the present invention can also reduce pain caused by dry eye, allergies, etc., resulting from the activation of the stimulatory receptor TRPA1. Therefore, as one embodiment of the present invention, an ophthalmic composition for suppressing pain caused by dry eye, allergies, etc., and a pain suppressant for dry eye, allergies, etc., are provided, which contain component (A). This is based on the fact that TRPA1 has also been reported to be involved in pain caused by dry eye (see, for example, Neuroscience. 2015 Apr 2;290:204-13). Furthermore, pain may be caused by allergies as well as dry eyes. Note that "dry eyes" includes not only dry eyes according to the dry eye diagnostic criteria, but also general dryness of the eyes.

[0061] [Irritant inhibitors for ophthalmic compositions] Furthermore, the present invention also provides an irritation inhibitor for ophthalmic compositions. The formulation and form of components (A) and (B) in the irritation inhibitor are as described above for ophthalmic compositions. One embodiment of the present invention provides an irritation inhibitor for an ophthalmic composition containing component (B), which contains component (A) as an active ingredient. When the TRPA1 agonist is menthol, it is possible to suppress irritation of the ophthalmic composition while maintaining a cooling sensation.

[0062] [Method for suppressing irritation from ophthalmic compositions] The present invention also provides a method for suppressing irritation of an ophthalmic composition, comprising incorporating component (A) and component (B) into the ophthalmic composition. Specifically, the method for suppressing irritation of an ophthalmic composition derived from (B) a TRPA1 agonist is characterized by further incorporating into an ophthalmic composition containing (B) a TRPA1 agonist, (A-1) an anticongestive component, (A-2) an antihistamine component, (A-3) at least one selected from the group consisting of camphor, menthone, isomenthol, cineole, mentofuran, pinene, limonene, neomenthol, and methyl acetate, and (A-4) a local anesthetic component.

[0063] [Methods for evaluating eye irritation] Furthermore, the present invention also provides a method for evaluating eye irritation. For example, a method for evaluating eye irritation caused by a test sample, characterized by contacting the test sample with TRPA1-expressing cells and measuring the change in intracellular calcium ion concentration caused by the test sample via TRPA1. As a test sample used in the evaluation method of the present invention, for example, a combination of (B)TRPA1 agonist and components commonly used in the field of ophthalmic compositions can be used. For example, such components may be the above-mentioned components (A-1), (A-2), (A-3), or (A-4), and may also be surfactants, thickeners, buffers, isotonic agents, carriers, chelating agents, bases, pH adjusters, stabilizers, preservatives, bactericides or antibacterial agents, sugars, sugar alcohols, ocular muscle regulating agents, vitamins, amino acids, anti-inflammatory agents, astringents, etc. Furthermore, (B)TRPA1 agonists may be used as a control sample (control sample) to the test sample. TRPA1-expressing cells are not particularly limited as long as they can stably retain the hTRPA1 gene, but for example, T-REx-293 cells and HEK293T cells are preferred. In the above method, the change in intracellular calcium ion concentration is, for example, known Ca 2+ The concentration indicators, such as Fluo-4, Fluo-8, or Fluo-2, can be used to measure and observe the levels using commercially available microplate readers or fluorescence microscopes. In this invention, by measuring the change in intracellular calcium ion concentration, information such as the presence or absence of eye irritation and the intensity of eye irritation can be obtained. Furthermore, by adjusting the time during which the test sample and control sample are in contact with TRPA1-expressing cells, intracellular calcium ion concentration can be observed over time, and information regarding the affinity of the test sample to TRPA1-expressing cells (for example, correlation with the LogP value (partition coefficient) of the test sample) can also be obtained. Furthermore, methods for bringing the test sample into contact with TRPA1-expressing cells include, for example, adding the test sample or its dilution, or artificial tears containing the test sample, to cultured TRPA1-expressing cells or animals such as humans, or administering them as eye drops.

[0064] The present invention will be described in more detail below with reference to examples, but the present invention is not limited in any way to these examples. [Examples]

[0065] <Cells used> Preparation of a T-REx-293 cell line containing the hTRPA1 gene hTRPA1 mRNA was extracted from human WI38 cells. HEK cells stably expressing hTRPA1 were created using Invitrogen's tetracycline-regulated T-REx expression system. hTRPA1 cDNA, amplified by RT-PCR using hTRPA1 mRNA as a template, was incorporated into the mammalian cell gene expression vector pcDNA4 / TO (Invitrogen) and introduced into T-REx-293 cells using Invitrogen's Lipofectamin 2000 reagent. pcDNA4 / TO contains Zeocin, and T-REx-293 cells retain a plasmid (blastosidine resistance gene) necessary for maintaining the T-Rex system. T-REx-293 cell lines stably retaining the hTRPA1 gene were established using the antibiotics Zeocin 500 μg / mL and blastosidine 10 μg / mL.

[0066] <Cell culture> T-REx-293 cells retaining hTRPA1 were cultured in DMEM medium containing 10% FBS, 100 units / mL penicillin, 100 mg / mL streptomysin, and 250 ng / mL amphotericin B, in the presence of 5% carbon dioxide. During subculturing, 200 μg / mL Zeocin and 5 μg / mL blastosidine were added, and the maximum number of subculturing was limited to 50. Specifically, as a pre-culture, T-REx-293 cells were cultured in a 60 mm dish to semi-confluent, and then washed with PBS(-). The cells were detached from the dish by 1×Trypsin / EDTA treatment, and DMEM medium was added. Then, the cells were transferred to a 15 mL tube and centrifuged at 800 rpm at room temperature for 4 minutes. Only the DMEM medium was aspirated and removed, the cells were loosened by tapping, and then fresh DMEM medium was added, resulting in approximately 40 × 10⁻⁶ cells. 4 ~50×10 4 A cell suspension was prepared by adjusting the cell count to achieve a cell / mL ratio. At this time, 1 μg / mL tetracycline was added to ensure receptor expression. The prepared cell suspension was dispensed in 100 μL portions into 96-well plates and incubated overnight at 37°C in a 5% CO2 incubator before being used for testing.

[0067] [Test Example 1] Confirmation test for suppression of irritation by l-menthol Menthol is known to stimulate the stimuli receptor TRPA1 by activating it. Therefore, l-menthol and the test sample (Example) were administered to the cells described above, and it was confirmed whether the test sample inhibited TRPA1 activity by l-menthol. Ca 2+ We observed whether TRP activity changed using a concentration indicator (Fluo-4 or Fluo-8 (manufactured by AAT Bioquest, Inc.)). 2+ The concentration indicator was pre-introduced into T-REx-293 cells that already contained hTRPA1, and the fluorescence intensity was measured using a microplate reader. Specifically, the following protocol was followed. A 96-well plate, which had been prepared the previous day (cell culture as described above), was removed from the incubator, and the DMEM medium was removed by decantation. The plates were gently washed with 100 μL / well of buffer (pH 7.4, 5.37 mM KCl, 0.44 mM KH2PO4, 137 mM NaCl, 0.34 mM Na2HPO4, 5.56 mM D-glucose, 20 mM HEPES, 1 mM CaCl2, 0.1% bovine serum albumin, 2.5 mM probenecid, remainder purified water), and 50 μL of the same buffer dissolved in 1.5 μM Fluo-8 was added to each well. The plates were then treated in a 37°C, 5% CO2 incubator for 1 hour. After incubator treatment, the same buffer containing Fluo-8 was removed by decantation, and the plates were gently washed with 100 μL / well of the same buffer. Furthermore, 180 μL / well of the same buffer was added, and measurement was started using a Flexstation II (Registered Trademark) or Flexstation III (Registered Trademark) microplate reader (Molecular Devices, Sunnyvale, CA, USA). 30 seconds after the start of microplate reader measurement, 20 μL / well of the test sample was added, and the maximum fluorescence value was measured between 5 and 120 seconds after addition. This value was defined as the fluorescence value.

[0068] The TRP activity (%) was calculated based on the following formula.

number

[0069] Comparative Example 2 (when l-menthol is administered alone) shows a result of 100%. Figure 1 shows the calculated TRP activity of the test samples listed in Table 1 below. Comparative Example 1 is a control (l-menthol-free buffer (pH 7.4, 5.37 mM KCl, 0.44 mM KH2PO4, 137 mM NaCl, 0.34 mM Na2HPO4, 5.56 mM D-glucose, 20 mM HEPES, 1 mM CaCl2, 0.1% bovine serum albumin, 2.5 mM probenecid, remainder purified water)), and Comparative Example 2 is prepared by dissolving l-menthol (TRPA1 agonist) at a concentration of 66.7 μM in the same buffer as Comparative Example 1. The example is prepared by dissolving l-menthol (66.7 μM) and the components listed in Table 1 at the concentrations listed in Table 2 in the same buffer as Comparative Example 1.

[0070] [Table 1]

[0071] [Table 2]

[0072] As shown in Figure 1, the ophthalmic composition containing (A-1) the decongestant components of the present invention, tetrahydrozoline hydrochloride and naphazoline hydrochloride; (A-2) the antihistamine components, chlorpheniramine maleate and diphenhydramine hydrochloride; (A-3) the components d-camphor, dl-camphor, (-)-menthone, (+)-isomenthol, 1,8-cineole, menthofuran, α-pinene, β-pinene, (R)-(+)-limonene, (S)-(-)-limonene, neomenthol, methyl acetate; and (A-4) the component chlorobutanol, was able to suppress the irritation caused by the TRPA1 agonist. Similarly, the ophthalmic composition containing essential oils containing component (A-3), namely peppermint oil and eucalyptus oil, was also able to suppress the irritation caused by the TRPA1 agonist.

[0073] [Test Example 2] Visualization, time-dependent observation, and evaluation of TRPA1 activation of l-menthol. Based on the same principle as in Test Example 1 above, Ca 2+ The degree of TRP activation was measured using Fluo-8, a calcium concentration indicator whose fluorescence intensity increases in accordance with calcium concentration. Ca 2+ The degree of TRP activation was measured using Fluo-8, a calcium concentration indicator whose fluorescence intensity increases in accordance with calcium concentration. Ca 2+ The calcium concentration indicator was preloaded into T-REx-293 cells harboring the hTRPA1 gene, and the fluorescence intensity was visualized using a fluorescence microscope system (Leica, LAS AF6000).

[0074] A T-REx-293 cell DMEM suspension containing the hTRPA1 gene at a concentration of about 60×10 4 cells / mL was added dropwise at 20 μL per slide glass onto a cover glass, and incubated overnight in a 37°C, 5% CO2 incubator. The incubated cover glass was removed from the incubator, and the medium was decanted off. The cover glass was gently washed with a buffer solution (pH 7.4, 5.37 mM KCl, 0.44 mM KH2PO4, 137 mM NaCl, 0.34 mM Na2HPO4, 5.56 mM D-glucose, 20 mM HEPES, 1 mM CaCl2, remainder purified water), then the fluorescence indicator Fluo-8 / buffer solution was added, and the mixture was treated in a 37°C, 5% CO2 incubator for 1 hour. After the incubator treatment, the cover glass was placed on a chamber, and the fluorescence intensity was visualized and observed over time using a fluorescence microscope system (Leica, LAS AF6000).

[0075] The flow of this test is as follows. (1) After confirming that the luminescence intensity of the fluorescent reagent is stable in the buffer solution, the buffer solution is replaced with a buffer solution containing l-menthol (66.7 μM) to activate TRPA1. (2) After the activation of TRPA1 is confirmed, the buffer solution containing l-menthol (66.7 μM) is replaced with a buffer solution containing the same concentration of l-menthol in combination with each test sample (tetrahydrozoline hydrochloride 0.003125 w / v%, peppermint oil 0.015 w / v%), to confirm whether TRPA1 activity is inhibited. (3) After confirming the inhibitory effect on TRPA1 activity, return the buffer containing l-menthol and each test sample to the l-menthol buffer used in (1) and check whether TRPA1 is reactivated.

[0076] Figure 2 shows the results of visualizing TRPA1 activation by l-menthol using a fluorescence microscope. When borneol was used as the test sample, it showed an inhibitory effect on TRPA1 activity, but when the test solution was returned to l-menthol, it showed TRPA1 activity again (see reference example in Figure 2). On the other hand, when peppermint oil, an essential oil containing (A-1) tetrahydrozoline hydrochloride, a decongestant component, and (A-3) component, was used as the test sample, it showed an inhibitory effect on TRPA1 activity, but when the test solution was returned to menthol only, no inhibitory effect was observed, or reactivation was weak (see example in Figure 2). In other words, it is thought that components (A-1) and (A-3) of the present invention were able to sustain their inhibitory effect on the TRPA1 receptor for a period of time.

[0077] [Test Example 3] Confirmation test of irritation suppression by tetrahydrozoline hydrochloride in humans Figure 3 shows the results of evaluating the cooling and irritation sensations when two solutions were administered to the left and right eyes of the same subject: a comparative example (Rohto Soft One Eye Drops, manufactured by Rohto Pharmaceutical Co., Ltd., containing 0.1 w / v% potassium chloride, 0.4 w / v% sodium chloride, boric acid, and a pH adjuster) in which l-menthol (0.01 w / v%) and polyoxyethylene hydrogenated castor oil (0.1 w / v%) were dissolved; and an example (the same artificial tear solution in which l-menthol (0.01 w / v%), polyoxyethylene hydrogenated castor oil (0.1 w / v%) and tetrahydrozoline hydrochloride (0.05 w / v%) were dissolved; respectively. The subjects were eight adult males, either with or without glasses, and the evaluations were quantified using a VAS scale (maximum scale: 20).

[0078] As a result, it was shown that by combining l-menthol with the (A-1) decongestant component, tetrahydrozoline hydrochloride, the cooling sensation of l-menthol can be maintained while suppressing the irritation caused by l-menthol. In other words, it was confirmed in humans that the (A-1) decongestant component of the present invention can suppress the irritation caused by TRPA1 agonists.

[0079] [Examples of formulations] The example formulations shown in Tables 4 and 5 below were prepared. Purified water was added to bring the total volume of each liquid to 100 mL.

[0080] [Table 4]

[0081] [Table 5] [Industrial applicability]

[0082] The present invention relates to an ophthalmic composition with suppressed irritation, an irritation inhibitor for ophthalmic compositions, a method for suppressing irritation from ophthalmic compositions, and a method for evaluating eye irritation. The agents of the present invention are useful in the field of pharmaceuticals.

Claims

1. Contains α-pinene, A composition for inhibiting the activity of the l-menthol-induced stimuli receptor TRPA1 by coexisting with l-menthol in an ophthalmic composition, A composition for ophthalmic use, for inhibiting the activity of the stimuli receptor TRPA1, characterized in that the α-pinene content is 0.0001 to 100 parts by weight per 1 part by weight of the total l-menthol content.

2. Contains menthone, A composition for inhibiting the activity of the l-menthol-induced stimuli receptor TRPA1 by coexisting with l-menthol in an ophthalmic composition, A composition for ophthalmic use, for inhibiting the activity of the stimuli receptor TRPA1, characterized in that the menthone content ratio is 0.0001 to 100 parts by weight per 1 part by weight of the total l-menthol content.

3. The ophthalmic composition containing l-menthol further includes a step of adding α-pinene, and A method for inhibiting the activity of the stimulatory receptor TRPA1, characterized in that the α-pinene content is 0.0001 to 100 parts by weight per 1 part by weight of the total l-menthol content.

4. The ophthalmic composition containing l-menthol further includes the step of containing menthone, and A method for inhibiting the activity of the stimulatory receptor TRPA1, characterized in that the menthone content ratio is 0.0001 to 100 parts by weight per 1 part by weight of the total l-menthol content.

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