WO2021261576A1 - Lipid secretion promoting agent - Google Patents
Lipid secretion promoting agent Download PDFInfo
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- WO2021261576A1 WO2021261576A1 PCT/JP2021/024074 JP2021024074W WO2021261576A1 WO 2021261576 A1 WO2021261576 A1 WO 2021261576A1 JP 2021024074 W JP2021024074 W JP 2021024074W WO 2021261576 A1 WO2021261576 A1 WO 2021261576A1
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- diquafosol
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- sodium
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7084—Compounds having two nucleosides or nucleotides, e.g. nicotinamide-adenine dinucleotide, flavine-adenine dinucleotide
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
Definitions
- the present invention relates to a lipid secretion-promoting agent containing diquafosol or a salt thereof (hereinafter, also simply referred to as "diquafosol") as an active ingredient, wherein the lipid secretion is a holocrine type.
- the present invention also relates to a tear film stabilizer containing the lipid secretion promoter.
- the present invention also relates to an ophthalmic composition containing diquafosol as an active ingredient for promoting lipid secretion, wherein the lipid secretion is a holocrine type.
- Dry eye is a disease that causes dry eye symptoms such as eye discomfort or visual dysfunction due to quantitative or qualitative abnormalities in tears, and may cause damage to the surface of the eye (cornea, etc.). .. Quantitative abnormalities mainly refer to a state in which the amount of tears secreted is low. On the other hand, qualitative abnormalities mainly refer to abnormalities in the tear film component, for example, the stability of the tear film layer is reduced due to a small amount of lipid component or protein component contained in the tear fluid, and tear fluid is secreted. Even if it is done, it can cause thirst on the surface of the eye.
- diquafosol is P 1, P 4 - di (uridine-5 ') and purine receptor agonist, also called tetraphosphate or Up4U, 3% (w / v ) concentration of Diquafosol tetrasodium salt (hereinafter, "Axis An ophthalmic solution containing (also referred to as “ahosol sodium”) is used as a therapeutic agent for dry eye (product name: Diquafosol (registered trademark) ophthalmic solution 3%).
- Axis Aho Sol sodium acts to P2Y 2 receptors on conjunctival epithelial and goblet cell membranes, by increasing the intracellular calcium concentration, it promotes the secretion of water and mucin (Jikuasu ® ophthalmic solution 3% Attachment (Non-Patent Document 1)).
- diquafosol sodium has a mucin-containing tear secretion promoting action and a mucin production promoting action of corneal epithelial cells (Non-Patent Document 1).
- the tear film is composed of three layers, a lipid layer (oil layer), an aqueous layer, and a mucin layer, and the outermost lipid layer prevents water from evaporating from the tear fluid, so that the tear film is stable. It is considered to be important for the conversion. Most of the lipids in tears are supplied as meibomian glands from the meibomian glands. However, even at present, there is no drug that improves the lipid layer (oil layer).
- the subject of the present invention is to elucidate a new mechanism of diquafosol and to provide a new agent.
- the present inventors have found that diquafosol acts on meibomian gland cells to promote holocrine lipid secretion, and completed the present invention.
- the present invention is based on surprising results, as no agent has been found to date that acts on meibomian gland cells to promote holocrine lipid secretion. Therefore, the present invention is useful for the prevention or treatment of eye diseases prevented or treated by promoting lipid secretion, eye diseases requiring lipid supplementation, eye diseases caused by lipid reduction, and the like.
- it is useful for the prevention or treatment of meibomian gland dysfunction (also referred to as MGD (meibomian gland dysfunction)), blepharitis, and dry eye (including dry eye caused by lipid loss).
- the present invention relates to the following.
- the lipid is at least one selected from the group consisting of wax esters, triglycerides, omega hydroxide lipids, cholesterol, cholesterol esters, total cholesterol, phospholipids, fatty acids and fatty alcohols, (1) or (2). ).
- the lipid secretion promoter is at least one selected from the group consisting of wax esters, triglycerides, omega hydroxide lipids, cholesterol, cholesterol esters, total cholesterol, phospholipids, fatty acids and fatty alcohols, (1) or (2). ).
- a tear layer stabilizer containing the lipid secretion promoter according to any one of (1) to (3).
- An ophthalmic composition which is at least one selected from the group consisting of cholesterol, cholesterol esters, total cholesterol, phospholipids, fatty acids and fatty alcohols.
- a preventive or therapeutic agent for dry eye containing the lipid secretion-promoting agent according to any one of (1) to (3).
- a method for preventing or treating dry eye which comprises administering to a patient the lipid secretion-promoting agent according to any one of (1) to (3).
- a preventive or therapeutic agent for dry eye which comprises the tear film stabilizer according to (4).
- a method for preventing or treating dry eye which comprises administering to a patient the tear film stabilizer according to (4).
- a method for preventing or treating dry eye which comprises administering to a patient the ophthalmic composition according to any one of (5) to (7).
- Diquafosol via the P2Y 2 receptor signaling has excellent lipid secretion promoting action because it enhances lipid secretion in meibomian gland cells, ocular disease to be prevented or treated by lipid secretagogue, require replenishment of lipids It is useful for the prevention or treatment of eye diseases caused by diquafosol and eye diseases caused by decreased lipids. For example, it is useful for the prevention or treatment of meibomian gland dysfunction (MGD), blepharitis, dry eye (including dry eye caused by lipid loss) and the like.
- MMD meibomian gland dysfunction
- blepharitis dry eye (including dry eye caused by lipid loss) and the like.
- FIG. 3A is a diagram showing the expression state of the apoptosis marker after the addition of diquafosol
- FIG. 3B is a diagram showing the amount of apoptosis generated after the addition of diquafosol.
- the lipid secretion-promoting agent of the present invention is a lipid secretion-promoting agent containing diquafosol or a salt thereof as an active ingredient, and is characterized by a holocrine type of lipid secretion.
- Diquafosol is a compound represented by the following chemical structural formula.
- the “salt of diquafosol” is not particularly limited as long as it is a pharmaceutically acceptable salt, and is a metal salt with lithium, sodium, potassium, calcium, magnesium, zinc, etc .; hydrochloric acid, hydrobromic acid, hydrogen iodide.
- Salts with inorganic acids such as nitrate, sulfuric acid, phosphoric acid; acetic acid, fumaric acid, maleic acid, succinic acid, citric acid, tartaric acid, adipic acid, gluconic acid, glucoheptic acid, glucuronic acid, terephthalic acid, methanesulfonic acid, Lactic acid, horse uric acid, 1,2-ethandisulfonic acid, isetionic acid, lactobionic acid, oleic acid, pamoic acid, polygalacturonic acid, stearic acid, tannic acid, trifluoromethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, Salts with organic acids such as lauryl sulfate, methyl sulfate, naphthalene sulfonic acid, sulfosalicylic acid; quaternary ammonium salts with methyl bromid
- diquafosol or a salt thereof also includes a hydrate and an organic solvate of diquafosol (free form) or a salt thereof.
- crystal polymorphs are present in “diquafosol or salts thereof"
- those polymorphs and polymorphs are also within the scope of the present invention.
- the crystal polymorph group is an individual crystal shape and its process at each stage when the crystal shape changes due to conditions and conditions such as production, crystallization, and storage of those crystals. Means the whole.
- the "diquafosol or a salt thereof" in the present invention is preferably a sodium salt of diquafosol, and a diquafosol tetrasodium salt represented by the following chemical structural formula is particularly preferable.
- Diquafosol or a salt thereof can be produced by the method disclosed in Japanese Patent Publication No. 2001-510484.
- diquafosol is also effective as a stabilizer for the tear film because it promotes the secretion of lipids.
- the present invention also provides a tear film stabilizer containing the above-mentioned lipid secretion promoter.
- the present invention also provides an ophthalmic composition containing diquafosol or a salt thereof as an active ingredient for promoting lipid secretion, wherein the lipid secretion is a holocrine type.
- the "ophthalmic composition” refers to a composition for use in the prevention and / or treatment of eye diseases and the like.
- the lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention may contain an active ingredient other than diquafosol or a salt thereof, or may contain diquafosol or a salt thereof as the only active ingredient. You can also.
- the concentration of diquafosol or a salt thereof is not particularly limited, but is preferably 0.0001 to 10% (w / v), and is preferably 0.001 to 10% (w / v), for example. ), More preferably 0.01 to 10% (w / v), even more preferably 0.1 to 10% (w / v), and even more preferably 1 to 10% (w / v). It is more preferably w / v), more preferably 1 to 5% (w / v), and particularly preferably 3% (w / v).
- lipid secretion promoter tear film stabilizer, and ophthalmic composition of the present invention may be appropriately changed according to the dosage form, the severity of the patient's symptoms to be administered, age, weight, judgment of the doctor, and the like.
- the number of eye drops is preferably 6 times a day, 5 times a day, 4 times a day, 3 times a day, 2 times a day or once a day, 6 times a day.
- 4 times a day, 3 times a day or 2 times a day is more preferable, 4 times a day, 3 times a day or 2 times a day is more preferable, and 3 times a day is particularly preferable.
- holocrine is one of the modes of release of secretions from cells, and means a mode in which glandular cells themselves are disrupted and various substances in the cells are released as secretory substances, and the release of cell contents.
- the accompanying cell death is characterized by a mode of programmed cell death such as apoptosis.
- Holocrine is also called holocrine or holocrine.
- promoting the secretion of lipids like holocrine can also be referred to as holocrine-type promotion of lipid secretion.
- the secreted lipid is not particularly limited, but for example, wax ester, triglyceride (also referred to as triacylglycerol), omega hydroxide lipid, cholesterol (also referred to as free cholesterol), cholesterol ester, total fatty acid. , Phosphoric lipids, fatty acids (eg, oleic acid, etc.), fatty alcohols and the like.
- the total of cholesterol ester and free cholesterol is total cholesterol (also referred to as TC (total cholesterol)).
- the present invention is, diquafosol via the P2Y 2 receptor signaling, based on the result of promoting lipid secretion in all secreted from meibomian gland cells. Therefore, the lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention are caused by eye diseases prevented or treated by promoting lipid secretion, eye diseases requiring lipid supplementation, and lipid reduction. It is useful for the prevention or treatment of eye diseases. Examples include, but are not limited to, meibomian gland dysfunction (MGD), blepharitis, dry eye (including dry eye due to lipid loss), and the like.
- MMD meibomian gland dysfunction
- blepharitis dry eye (including dry eye due to lipid loss)
- Dry eye is defined as "a chronic disease of tears and keratoconjunctival epithelium caused by various factors and is accompanied by eye discomfort and visual abnormalities", and keratoconjunctivitis sicca (KCS) is included in dry eye. .. In the present invention, the occurrence of dry eye symptoms caused by wearing soft contact lenses is also included in dry eye.
- Dry eye symptoms include subjective symptoms such as dry eyes, discomfort, eye fatigue, dullness, dullness, eye pain, and blurred vision (blurred vision), as well as other symptoms such as congestion and keratoconjunctival epithelial disorders. Findings are also included.
- meibomian gland dysfunction is, for example, "a state in which the function of the meibomian glands is diffusely abnormal due to various causes, accompanied by chronic eye discomfort.”
- the "state in which the function of the meibomian gland is diffusely abnormal” is not a local meibomian gland abnormality observed in, for example, chalazion or hordeolum, but is a dilation of the meibomian gland or obstruction of the opening of the meibomian gland. It means that meibomian gland abnormalities such as are found diffusely.
- the MGD is divided into a secretion-reducing MGD (low-delivery state) and a secretion-increasing MGD (high-delivery state), and further as a secretion-reducing MGD (low-delivery state MD), "hyposectory MGD (meibomian gland) meibomian gland". "And" meibomian gland obstruction ".
- the secretion of meibomian gland fat is reduced due to obstruction of the meibomian gland opening.
- the secretion of meibomian gland fat increases due to various causes.
- MGD meibomian gland concretion
- meibomian gland concretion a state in which obstruction of the meibomian gland opening is observed but not accompanied by subjective symptoms
- MGD in the present invention refers to meibomian gland infarction. Is also included.
- MGD that diquafosol can treat is a hyposecretory MGD.
- MGD may cause dry eye and may also cause posterior blepharitis.
- MGD includes “MGD with (combined) dry eye and / or posterior blepharitis”, “MGD causing dry eye and / or posterior blepharitis”, “MGD without (uncomplicated) dry eye”. , “MGD that does not cause dry eye”, “MGD that does not accompany (do not combine) with posterior blepharitis” and “MGD that does not cause posterior blepharitis” are included.
- Blepharitis is a symptom of inflamed eyelids and includes anterior blepharitis, blepharitis, posterior blepharitis, blepharitis, and blepharitis.
- the lipid secretion promoter, tear layer stabilizer, and ophthalmic composition of the present invention can be used for in vivo administration.
- the lipid secretion promoter, tear layer stabilizer, and ophthalmic composition of the present invention can be used in an in vivo treatment method.
- the lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention are preferably used for the prevention and / or treatment of eye diseases and the like.
- Examples of the dosage form of the lipid secretion promoter, the tear film stabilizer, and the ophthalmic composition of the present invention include eye drops, eye ointments, injections, ointments (for example, which can be administered to the eyelid skin) and the like. It is preferably an eye drop.
- eye drops are synonymous with eye drops or eye drops, and eye drops for contact lenses are also included in the definition of eye drops.
- the lipid secretion promoter, tear layer stabilizer, and ophthalmic composition of the present invention preferably use water as a solvent (base), and more preferably an aqueous eye drop.
- the lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention may be a soluble eye drop or a suspension type eye drop depending on the properties and contents of the active ingredient and the additive. May be.
- the lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention can be further added with pharmaceutically acceptable additives as needed by using a general-purpose technique.
- pharmaceutically acceptable additives such as sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium acetate, epsilon-aminocaproic acid; isotonic agents such as calcium chloride, sodium chloride, potassium chloride, concentrated glycerin; edetonic acid.
- Stabilizers such as sodium; Surface active agents such as polysorbate; Antioxidants such as ascorbic acid; Preservatives such as benzalkonium chloride and chlorhexizing luconate; pH adjusters such as hydrochloric acid and sodium hydroxide are required. It can be selected and added accordingly. These additives may be used alone or in any combination of two or more.
- the pH of the lipid secretion promoter, the tear film stabilizer, and the ophthalmic composition of the present invention is not limited to a specific value as long as it is within the range acceptable for pharmaceuticals.
- the pH of the lipid secretion promoter, the tear film stabilizer, and the ophthalmic composition of the present invention is preferably 8 or less, more preferably in the range of 4 to 8, still more preferably in the range of 5 to 8, and even more. It is preferably in the range of 6 to 8, particularly preferably in the vicinity of 7.
- the present invention also provides an ophthalmic composition containing diquafosol or a salt thereof for use in an in vivo treatment method for promoting holocrine lipid secretion from meibomian gland cells.
- the present invention further comprises an ophthalmic composition comprising diquafosol or a salt thereof for use in an in vivo treatment method for promoting lipid secretion from mybome gland cells, wherein the lipid is a wax ester, triglyceride, omega water. Also provided is an ophthalmic composition which is at least one selected from the group consisting of lipid oxides, cholesterol, cholesterol esters, total cholesterol, phospholipids, fatty acids and fatty alcohols.
- Diquafosol sodium is dissolved in purified water to prepare an 8.5% (w / v) aqueous solution, which is then diluted with Ham F-12 to the desired concentration of diquafosol sodium (hereinafter abbreviated as "DQS"). ) Liquid was prepared.
- Diquafosol sodium is dissolved in purified water to prepare an 8.5% (w / v) aqueous solution, which is then diluted with Ham F-12 to the desired concentration of diquafosol sodium (hereinafter abbreviated as "DQS"). ) Liquid was prepared.
- Meibomian gland cells were isolated from white rabbits by enzyme treatment and cultured without serum in Ham F-12 medium containing various culture additives. After inducing cell differentiation, the cells were washed and various concentrations of diquafosol sodium (final concentrations: 0.03%, 0.3%, 0.85%) were added. Note in the group applying the P2Y 2 receptor antagonist AR-C118925XX (hereinafter generally as "AR-C", final concentration: 3 [mu] M) was also added at the same time. After incubation for 4 hours, the culture supernatant and cells were collected separately.
- AR-C P2Y 2 receptor antagonist
- the culture supernatant sample was left as it was, while the cell sample was subjected to extraction treatment of lipid components, and then the total cholesterol (TC) content was measured using a total cholesterol quantifying reagent (Amplex Red Cholesterol Assay, Invitrogen).
- sample preparation Dissolve diquafosol sodium in purified water to prepare an 8.5% (w / v) aqueous solution, and 1 volume of this diquafosol sodium (hereinafter abbreviated as "DQS") solution for 9 volumes of the culture solution. was added and used in the ratio of.
- DQS diquafosol sodium
- Meibomian gland cells were isolated from white rabbits by enzyme treatment and cultured without serum in Ham F-12 medium containing various culture additives. After inducing cell differentiation, the cells were washed, DQS solution was added, and the cells were incubated for 2 to 6 hours. The cells were collected together with the culture supernatant, and each fragmented DNA sample was obtained using ApopRadder Ex (Takara Bio Inc.). Fragmented DNA samples were electrophoresed on a 1.5% agarose gel and then stained with SYBR Green I dye to visualize DNA bands. At the same time, SYBR Green I dye was added to the fragmented DNA sample, the fluorescence intensity was measured, and the amount of fragmented DNA was quantified.
- FIG. 3A a ladder-shaped DNA band characteristic of apoptotic cells was detected in the fragmented DNA prepared from cultured meibomian gland cells. Moreover, this characteristic DNA ladder pattern was not disturbed even after the addition of DQS.
- FIG. 3 (b) the amount of fragmented DNA increased significantly 4 hours after the addition of DQS. That is, it was shown that the apoptotic cell death constitutively occurring in meibomian gland cells is promoted after the addition of DQS.
- each formulation such as eye drops can be prepared.
- Diquafosol or a salt thereof via the P2Y 2 receptor signaling has excellent lipid secretion promoting action because it enhances lipid secretion in meibomian gland cells, ocular disease to be prevented or treated by lipid secretagogue, recruitment of lipids It is useful for the prevention or treatment of eye diseases that require meibomian glands, eye diseases caused by lipid reduction, and the like.
Abstract
A promoting agent for the secretion of a lipid, which contains diquafosol or a salt thereof as an active ingredient, in which the type of the secretion of the lipid is of a holocrine type. An ophthalmic composition containing diquafosol or a salt thereof as an active ingredient, which is intended to be used for the promotion of the secretion of a lipid, in which the type of the secretion of the lipid is of a holocrine type. An ophthalmic composition containing diquafosol or a salt thereof, which is intended to be used in an in vivo treatment method for promoting the holocrine-type secretion of a lipid from a meibomian gland cell. An ophthalmic composition containing diquafosol or a salt thereof, which is intended to be used in an in vivo treatment method for promoting the secretion of a lipid from a meibomian gland cell, in which the lipid is at least one component selected from the group consisting of a wax ester, a triglyceride, an omega-hydroxy lipid, cholesterol, a cholesterol ester, total cholesterol, a phospholipid, a fatty acid and a fatty alcohol.
Description
本発明は、ジクアホソルまたはその塩（以下、単に「ジクアホソル」ともいう）を有効成分として含有する脂質分泌促進剤であって、脂質分泌が全分泌型である、脂質分泌促進剤に関する。また、本発明は、当該脂質分泌促進剤を含有する涙液層安定化剤にも関する。さらに本発明は、脂質分泌を促進するための、ジクアホソルを有効成分として含有する眼科用組成物であって、脂質分泌が全分泌型である、眼科用組成物にも関する。
The present invention relates to a lipid secretion-promoting agent containing diquafosol or a salt thereof (hereinafter, also simply referred to as "diquafosol") as an active ingredient, wherein the lipid secretion is a holocrine type. The present invention also relates to a tear film stabilizer containing the lipid secretion promoter. Furthermore, the present invention also relates to an ophthalmic composition containing diquafosol as an active ingredient for promoting lipid secretion, wherein the lipid secretion is a holocrine type.
ドライアイは、涙液の量的又は質的な異常に伴い、眼の不快感又は視機能異常などのドライアイ症状を生じる疾患であり、眼の表面（角膜など）に障害を引き起こすことがある。量的な異常は、主に涙液の分泌量が少ない状態をいう。一方、質的な異常は、主に涙液成分の異常を指し、例えば涙液に含まれる脂質成分又はタンパク質成分が少ないなどの理由により、涙液層の安定性が低下し、涙液が分泌されていても眼の表面の渇きをもたらしうる。
Dry eye is a disease that causes dry eye symptoms such as eye discomfort or visual dysfunction due to quantitative or qualitative abnormalities in tears, and may cause damage to the surface of the eye (cornea, etc.). .. Quantitative abnormalities mainly refer to a state in which the amount of tears secreted is low. On the other hand, qualitative abnormalities mainly refer to abnormalities in the tear film component, for example, the stability of the tear film layer is reduced due to a small amount of lipid component or protein component contained in the tear fluid, and tear fluid is secreted. Even if it is done, it can cause thirst on the surface of the eye.
一方、ジクアホソルはＰ１，Ｐ４－ジ（ウリジン－５’）四リン酸またはＵｐ４Ｕとも呼ばれるプリン受容体アゴニストであり、３％（ｗ／ｖ）の濃度のジクアホソル四ナトリウム塩（以下、「ジクアホソルナトリウム」ともいう）を含有する点眼液がドライアイ治療薬として使用されている（製品名：ジクアス（登録商標）点眼液３％）。ジクアホソルナトリウムは、結膜上皮および杯細胞膜上のＰ２Ｙ２受容体に作用し、細胞内のカルシウム濃度を上昇させることにより、水分及びムチンの分泌を促進する（ジクアス（登録商標）点眼液３％添付文書（非特許文献１））。また、ジクアホソルナトリウムは、ムチンを含む涙液分泌促進作用および角膜上皮細胞のムチン産生促進作用を有する（非特許文献１）。
Meanwhile, diquafosol is P 1, P 4 - di (uridine-5 ') and purine receptor agonist, also called tetraphosphate or Up4U, 3% (w / v ) concentration of Diquafosol tetrasodium salt (hereinafter, "Axis An ophthalmic solution containing (also referred to as "ahosol sodium") is used as a therapeutic agent for dry eye (product name: Diquafosol (registered trademark) ophthalmic solution 3%). Axis Aho Sol sodium acts to P2Y 2 receptors on conjunctival epithelial and goblet cell membranes, by increasing the intracellular calcium concentration, it promotes the secretion of water and mucin (Jikuasu ® ophthalmic solution 3% Attachment (Non-Patent Document 1)). In addition, diquafosol sodium has a mucin-containing tear secretion promoting action and a mucin production promoting action of corneal epithelial cells (Non-Patent Document 1).
ところで、涙液層は脂質層（油層）、水層、ムチン層の三層から構成されており、最も外側にある脂質層は涙液からの水の蒸発を防止することから涙液層の安定化に重要であると考えられている。涙液中のほとんどの脂質はマイボーム腺からマイバムとして供給される。しかしながら、現在においても脂質層（油層）を改善する薬剤は存在していない。
By the way, the tear film is composed of three layers, a lipid layer (oil layer), an aqueous layer, and a mucin layer, and the outermost lipid layer prevents water from evaporating from the tear fluid, so that the tear film is stable. It is considered to be important for the conversion. Most of the lipids in tears are supplied as meibomian glands from the meibomian glands. However, even at present, there is no drug that improves the lipid layer (oil layer).
本発明の課題は、ジクアホソルの新たなメカニズムを解明し、新たな剤を提供することである。
The subject of the present invention is to elucidate a new mechanism of diquafosol and to provide a new agent.
本発明者らは、鋭意研究した結果、ジクアホソルがマイボーム腺細胞に作用して、全分泌型で脂質分泌を促進することを見出し、本発明を完成させた。マイボーム腺細胞に作用して全分泌型で脂質分泌を促進する薬剤が今日まで見い出されていなかったことからすれば本発明は驚くべき結果に基づくものである。よって、本発明は、脂質分泌促進によって予防または治療される眼疾患、脂質の補充を必要とする眼疾患、脂質減少に起因する眼疾患などの予防または治療に有用である。例えば、マイボーム腺機能不全（ＭＧＤ（ｍｅｉｂｏｍｉａｎ ｇｌａｎｄ ｄｙｓｆｕｎｃｔｉｏｎ）ともいう）、眼瞼炎、ドライアイ（脂質減少に起因するドライアイを含む）などの予防または治療に有用である。
As a result of diligent research, the present inventors have found that diquafosol acts on meibomian gland cells to promote holocrine lipid secretion, and completed the present invention. The present invention is based on surprising results, as no agent has been found to date that acts on meibomian gland cells to promote holocrine lipid secretion. Therefore, the present invention is useful for the prevention or treatment of eye diseases prevented or treated by promoting lipid secretion, eye diseases requiring lipid supplementation, eye diseases caused by lipid reduction, and the like. For example, it is useful for the prevention or treatment of meibomian gland dysfunction (also referred to as MGD (meibomian gland dysfunction)), blepharitis, and dry eye (including dry eye caused by lipid loss).
すなわち、本発明は以下に関する。
（１）ジクアホソルまたはその塩を有効成分として含有する脂質分泌促進剤であって、脂質分泌が全分泌型である、脂質分泌促進剤。 That is, the present invention relates to the following.
(1) A lipid secretion-promoting agent containing diquafosol or a salt thereof as an active ingredient, wherein the lipid secretion is a holocrine type.
（１）ジクアホソルまたはその塩を有効成分として含有する脂質分泌促進剤であって、脂質分泌が全分泌型である、脂質分泌促進剤。 That is, the present invention relates to the following.
(1) A lipid secretion-promoting agent containing diquafosol or a salt thereof as an active ingredient, wherein the lipid secretion is a holocrine type.
（２）脂質分泌がマイボーム腺細胞からの分泌である、（１）に記載の脂質分泌促進剤。
(2) The lipid secretion-promoting agent according to (1), wherein lipid secretion is secretion from meibomian gland cells.
（３）脂質が、ワックスエステル、トリグリセリド、オメガ水酸化脂質、コレステロール、コレステロールエステル、総コレステロール、リン脂質、脂肪酸および脂肪アルコールからなる群から選択される少なくとも１つである、（１）または（２）に記載の脂質分泌促進剤。
(3) The lipid is at least one selected from the group consisting of wax esters, triglycerides, omega hydroxide lipids, cholesterol, cholesterol esters, total cholesterol, phospholipids, fatty acids and fatty alcohols, (1) or (2). ). The lipid secretion promoter.
（４）（１）～（３）のいずれか１に記載の脂質分泌促進剤を含有する、涙液層安定化剤。
(4) A tear layer stabilizer containing the lipid secretion promoter according to any one of (1) to (3).
（５）脂質分泌を促進するための、ジクアホソルまたはその塩を有効成分として含有する眼科用組成物であって、脂質分泌が全分泌型である、眼科用組成物。
(5) An ophthalmic composition containing diquafosol or a salt thereof as an active ingredient for promoting lipid secretion, wherein the lipid secretion is a holocrine type.
（６）マイボーム腺細胞からの全分泌型の脂質分泌を促進するためのインビボ処理方法において使用するための、ジクアホソルまたはその塩を含有する眼科用組成物。
(6) An ophthalmic composition containing diquafosol or a salt thereof for use in an in vivo treatment method for promoting holocrine lipid secretion from meibomian gland cells.
（７）マイボーム腺細胞からの脂質分泌を促進するためのインビボ処理方法において使用するための、ジクアホソルまたはその塩を含有する眼科用組成物であって、該脂質がワックスエステル、トリグリセリド、オメガ水酸化脂質、コレステロール、コレステロールエステル、総コレステロール、リン脂質、脂肪酸および脂肪アルコールからなる群から選択される少なくとも１つである、眼科用組成物。
(7) An ophthalmic composition containing diquafosol or a salt thereof for use in an in vivo treatment method for promoting lipid secretion from Mybome gland cells, wherein the lipid is a wax ester, triglyceride, or omega hydroxide lipid. An ophthalmic composition which is at least one selected from the group consisting of cholesterol, cholesterol esters, total cholesterol, phospholipids, fatty acids and fatty alcohols.
（８）（１）～（３）のいずれか１に記載の脂質分泌促進剤を含有する、ドライアイの予防または治療剤。
(8) A preventive or therapeutic agent for dry eye containing the lipid secretion-promoting agent according to any one of (1) to (3).
（９）患者に、（１）～（３）のいずれか１に記載の脂質分泌促進剤を投与することを含む、ドライアイの予防または治療方法。
(9) A method for preventing or treating dry eye, which comprises administering to a patient the lipid secretion-promoting agent according to any one of (1) to (3).
（１０）ドライアイの予防または治療に使用するための、（１）～（３）のいずれか１に記載の脂質分泌促進剤。
(10) The lipid secretion-promoting agent according to any one of (1) to (3) for use in the prevention or treatment of dry eye.
（１１）ドライアイを予防または治療するための医薬を製造するための、（１）～（３）のいずれか１に記載の脂質分泌促進剤の使用。
(11) Use of the lipid secretion-promoting agent according to any one of (1) to (3) for producing a drug for preventing or treating dry eye.
（１２）（４）に記載の涙液層安定化剤を含有する、ドライアイの予防または治療剤。
（１３）患者に、（４）に記載の涙液層安定化剤を投与することを含む、ドライアイの予防または治療方法。 (12) A preventive or therapeutic agent for dry eye, which comprises the tear film stabilizer according to (4).
(13) A method for preventing or treating dry eye, which comprises administering to a patient the tear film stabilizer according to (4).
（１３）患者に、（４）に記載の涙液層安定化剤を投与することを含む、ドライアイの予防または治療方法。 (12) A preventive or therapeutic agent for dry eye, which comprises the tear film stabilizer according to (4).
(13) A method for preventing or treating dry eye, which comprises administering to a patient the tear film stabilizer according to (4).
（１４）ドライアイの予防または治療に使用するための、（４）に記載の涙液層安定化剤。
(14) The tear film stabilizer according to (4) for use in the prevention or treatment of dry eye.
（１５）ドライアイを予防または治療するための医薬を製造するための、（４）に記載の涙液層安定化剤の使用。
(15) Use of the tear film stabilizer according to (4) for producing a drug for preventing or treating dry eye.
（１６）（５）～（７）のいずれか１に記載の眼科用組成物を含有する、ドライアイの予防または治療剤。
(16) A preventive or therapeutic agent for dry eye containing the ophthalmic composition according to any one of (5) to (7).
（１７）患者に、（５）～（７）のいずれか１に記載の眼科用組成物を投与することを含む、ドライアイの予防または治療方法。
(17) A method for preventing or treating dry eye, which comprises administering to a patient the ophthalmic composition according to any one of (5) to (7).
（１８）ドライアイの予防または治療に使用するための、（５）～（７）のいずれか１に記載の眼科用組成物。
(18) The ophthalmic composition according to any one of (5) to (7) for use in the prevention or treatment of dry eye.
（１９）ドライアイを予防または治療するための医薬を製造するための、（５）～（７）のいずれか１に記載の眼科用組成物の使用。
(19) Use of the ophthalmic composition according to any one of (5) to (7) for producing a drug for preventing or treating dry eye.
ジクアホソルはＰ２Ｙ２受容体シグナル伝達を介して、マイボーム腺細胞における脂質分泌を増強することから優れた脂質分泌促進作用を有し、脂質分泌促進によって予防または治療される眼疾患、脂質の補充を必要とする眼疾患、脂質減少に起因する眼疾患などの予防または治療に有用である。例えば、マイボーム腺機能不全（ＭＧＤ）、眼瞼炎、ドライアイ（脂質減少に起因するドライアイを含む）などの予防または治療に有用である。
Diquafosol via the P2Y 2 receptor signaling, has excellent lipid secretion promoting action because it enhances lipid secretion in meibomian gland cells, ocular disease to be prevented or treated by lipid secretagogue, require replenishment of lipids It is useful for the prevention or treatment of eye diseases caused by diquafosol and eye diseases caused by decreased lipids. For example, it is useful for the prevention or treatment of meibomian gland dysfunction (MGD), blepharitis, dry eye (including dry eye caused by lipid loss) and the like.
本発明についてさらに詳しく説明する。
本発明の脂質分泌促進剤は、ジクアホソルまたはその塩を有効成分として含有する脂質分泌促進剤であって、脂質分泌が全分泌型であることを特徴とする。「ジクアホソル」は、下記化学構造式で示される化合物である。 The present invention will be described in more detail.
The lipid secretion-promoting agent of the present invention is a lipid secretion-promoting agent containing diquafosol or a salt thereof as an active ingredient, and is characterized by a holocrine type of lipid secretion. "Diquafosol" is a compound represented by the following chemical structural formula.
本発明の脂質分泌促進剤は、ジクアホソルまたはその塩を有効成分として含有する脂質分泌促進剤であって、脂質分泌が全分泌型であることを特徴とする。「ジクアホソル」は、下記化学構造式で示される化合物である。 The present invention will be described in more detail.
The lipid secretion-promoting agent of the present invention is a lipid secretion-promoting agent containing diquafosol or a salt thereof as an active ingredient, and is characterized by a holocrine type of lipid secretion. "Diquafosol" is a compound represented by the following chemical structural formula.
「ジクアホソルの塩」としては、医薬として許容される塩であれば特に制限はなく、リチウム、ナトリウム、カリウム、カルシウム、マグネシウム、亜鉛などとの金属塩；塩酸、臭化水素酸、ヨウ化水素酸、硝酸、硫酸、リン酸などの無機酸との塩；酢酸、フマル酸、マレイン酸、コハク酸、クエン酸、酒石酸、アジピン酸、グルコン酸、グルコヘプト酸、グルクロン酸、テレフタル酸、メタンスルホン酸、乳酸、馬尿酸、１，２－エタンジスルホン酸、イセチオン酸、ラクトビオン酸、オレイン酸、パモ酸、ポリガラクツロン酸、ステアリン酸、タンニン酸、トリフルオロメタンスルホン酸、ベンゼンスルホン酸、ｐ－トルエンスルホン酸、硫酸ラウリルエステル、硫酸メチル、ナフタレンスルホン酸、スルホサリチル酸などの有機酸との塩；臭化メチル、ヨウ化メチルなどとの四級アンモニウム塩；臭素イオン、塩素イオン、ヨウ素イオンなどのハロゲンイオンとの塩；アンモニアとの塩；トリエチレンジアミン、２－アミノエタノール、２，２－イミノビス（エタノール）、１－デオキシ－１－（メチルアミノ）－２－Ｄ－ソルビトール、２－アミノ－２－（ヒドロキシメチル）－１，３－プロパンジオール、プロカイン、Ｎ，Ｎ－ビス（フェニルメチル）－１，２－エタンジアミンなどの有機アミンとの塩などが挙げられる。
The "salt of diquafosol" is not particularly limited as long as it is a pharmaceutically acceptable salt, and is a metal salt with lithium, sodium, potassium, calcium, magnesium, zinc, etc .; hydrochloric acid, hydrobromic acid, hydrogen iodide. , Salts with inorganic acids such as nitrate, sulfuric acid, phosphoric acid; acetic acid, fumaric acid, maleic acid, succinic acid, citric acid, tartaric acid, adipic acid, gluconic acid, glucoheptic acid, glucuronic acid, terephthalic acid, methanesulfonic acid, Lactic acid, horse uric acid, 1,2-ethandisulfonic acid, isetionic acid, lactobionic acid, oleic acid, pamoic acid, polygalacturonic acid, stearic acid, tannic acid, trifluoromethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, Salts with organic acids such as lauryl sulfate, methyl sulfate, naphthalene sulfonic acid, sulfosalicylic acid; quaternary ammonium salts with methyl bromide, methyl iodide, etc .; with halogen ions such as bromine ion, chlorine ion, iodine ion Salt; Salt with ammonia; Triethylenediamine, 2-aminoethanol, 2,2-iminobis (ethanol), 1-deoxy-1- (methylamino) -2-D-sorbitol, 2-amino-2- (hydroxymethyl) )-1,3-Propanediol, prokine, N, N-bis (phenylmethyl) -1,2-ethanediamine and other salts with organic amines.
本発明において、「ジクアホソルまたはその塩」には、ジクアホソル（フリー体）またはその塩の水和物および有機溶媒和物も含まれる。
In the present invention, "diquafosol or a salt thereof" also includes a hydrate and an organic solvate of diquafosol (free form) or a salt thereof.
「ジクアホソルまたはその塩」に、結晶多形および結晶多形群（結晶多形システム）が存在する場合には、それらの結晶多形体および結晶多形群（結晶多形システム）も本発明の範囲に含まれる。ここで、結晶多形群（結晶多形システム）とは、それらの結晶の製造、晶出、保存などの条件および状態により、結晶形が変化する場合の各段階における個々の結晶形およびその過程全体を意味する。
If polymorphs and polymorphs (crystal polymorphs) are present in "diquafosol or salts thereof", those polymorphs and polymorphs (polymorphs) are also within the scope of the present invention. include. Here, the crystal polymorph group (crystal polymorph system) is an individual crystal shape and its process at each stage when the crystal shape changes due to conditions and conditions such as production, crystallization, and storage of those crystals. Means the whole.
本発明における「ジクアホソルまたはその塩」として好ましいのはジクアホソルのナトリウム塩であり、下記化学構造式で示されるジクアホソル四ナトリウム塩が、特に好ましい。
The "diquafosol or a salt thereof" in the present invention is preferably a sodium salt of diquafosol, and a diquafosol tetrasodium salt represented by the following chemical structural formula is particularly preferable.
ジクアホソルまたはその塩については、特表２００１－５１０４８４号公報に開示された方法などにより製造することができる。
Diquafosol or a salt thereof can be produced by the method disclosed in Japanese Patent Publication No. 2001-510484.
後述の試験例でも示すように、ジクアホソルは、脂質の分泌を促進することから、涙液層の安定化剤としても有効である。本発明は、上述した脂質分泌促進剤を含有する、涙液層安定化剤についても提供する。
As shown in the test examples described later, diquafosol is also effective as a stabilizer for the tear film because it promotes the secretion of lipids. The present invention also provides a tear film stabilizer containing the above-mentioned lipid secretion promoter.
また本発明は脂質分泌を促進するための、ジクアホソルまたはその塩を有効成分として含有する眼科用組成物であって、脂質分泌が全分泌型である、眼科用組成物についても提供する。本発明において、「眼科用組成物」とは、眼疾患などの予防および／または治療に使用するための組成物のことをいう。
The present invention also provides an ophthalmic composition containing diquafosol or a salt thereof as an active ingredient for promoting lipid secretion, wherein the lipid secretion is a holocrine type. In the present invention, the "ophthalmic composition" refers to a composition for use in the prevention and / or treatment of eye diseases and the like.
本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物は、ジクアホソルまたはその塩以外の有効成分を含有することもできるし、ジクアホソルまたはその塩を唯一の有効成分として含有することもできる。
The lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention may contain an active ingredient other than diquafosol or a salt thereof, or may contain diquafosol or a salt thereof as the only active ingredient. You can also.
本発明において、ジクアホソルまたはその塩の濃度は、特に限定されるものではないが、例えば、０．０００１～１０％（ｗ／ｖ）であることが好ましく、０．００１～１０％（ｗ／ｖ）であることがより好ましく、０．０１～１０％（ｗ／ｖ）であることがさらに好ましく、０．１～１０％（ｗ／ｖ）であることがよりさらに好ましく、１～１０％（ｗ／ｖ）であることがもっと好ましく、１～５％（ｗ／ｖ）であることがことさら好ましく、３％（ｗ／ｖ）であることが特に好ましい。
In the present invention, the concentration of diquafosol or a salt thereof is not particularly limited, but is preferably 0.0001 to 10% (w / v), and is preferably 0.001 to 10% (w / v), for example. ), More preferably 0.01 to 10% (w / v), even more preferably 0.1 to 10% (w / v), and even more preferably 1 to 10% (w / v). It is more preferably w / v), more preferably 1 to 5% (w / v), and particularly preferably 3% (w / v).
本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物の用法は、剤型、投与すべき患者の症状の軽重、年齢、体重、医師の判断などに応じて適宜変えることができるが、例えば、剤型として点眼剤を選択した場合には、１回量１～５滴、好ましくは１～３滴、より好ましくは１～２滴、特に好ましくは１滴を、１日１～６回、好ましくは１日１～４回、より好ましくは１日２～４回、さらに好ましくは１日３回、毎日～１週間毎に点眼投与することができる。ここで点眼回数は、より具体的には、例えば、１日６回、１日５回、１日４回、１日３回、１日２回または１日１回が好ましく、１日６回、１日４回、１日３回または１日２回がより好ましく、１日４回、１日３回または１日２回がさらに好ましく、１日３回が特に好ましい。
The usage of the lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention may be appropriately changed according to the dosage form, the severity of the patient's symptoms to be administered, age, weight, judgment of the doctor, and the like. However, for example, when an eye drop is selected as the dosage form, a single dose of 1 to 5 drops, preferably 1 to 3 drops, more preferably 1 to 2 drops, particularly preferably 1 drop, 1 drop per day. It can be administered by eye drops to 6 times, preferably 1 to 4 times a day, more preferably 2 to 4 times a day, still more preferably 3 times a day, every day to 1 week. Here, more specifically, for example, the number of eye drops is preferably 6 times a day, 5 times a day, 4 times a day, 3 times a day, 2 times a day or once a day, 6 times a day. 4 times a day, 3 times a day or 2 times a day is more preferable, 4 times a day, 3 times a day or 2 times a day is more preferable, and 3 times a day is particularly preferable.
後述の試験例でも示すように、ジクアホソルは、マイボーム腺細胞（マイボサイトともいう）に作用して、全分泌様脂質の分泌を促進する。ここで全分泌とは、細胞からの分泌物の放出様式の１つで、腺細胞自体が崩壊し、細胞内の様々な物質が分泌物質として放出される様式を意味し、細胞内容物の放出に伴って生ずる細胞死は、アポトーシスなどのプログラム細胞死の様式であることがその特徴であるとされる。全分泌は、ホロクリンやホロクラインとも呼ばれる。なお、全分泌様に脂質の分泌を促進することを全分泌型の脂質分泌促進ともいうことができる。
As shown in the test examples described later, diquafosol acts on meibomian gland cells (also called meibomian glands) to promote the secretion of holocrine-like lipids. Here, holocrine is one of the modes of release of secretions from cells, and means a mode in which glandular cells themselves are disrupted and various substances in the cells are released as secretory substances, and the release of cell contents. The accompanying cell death is characterized by a mode of programmed cell death such as apoptosis. Holocrine is also called holocrine or holocrine. In addition, promoting the secretion of lipids like holocrine can also be referred to as holocrine-type promotion of lipid secretion.
本発明において、分泌される脂質は特に限定されるものではないが、例えば、ワックスエステル、トリグリセリド（トリアシルグリセロールともいう）、オメガ水酸化脂質、コレステロール（遊離コレステロールともいう）、コレステロールエステル、総コレステロール、リン脂質、脂肪酸（例えば、オレイン酸など）、脂肪アルコールなどが挙げられる。なお、コレステロールエステルと遊離コレステロールの総和が総コレステロール（ＴＣ（ｔｏｔａｌ ｃｈｏｌｅｓｔｅｒｏｌ）ともいう）である。
In the present invention, the secreted lipid is not particularly limited, but for example, wax ester, triglyceride (also referred to as triacylglycerol), omega hydroxide lipid, cholesterol (also referred to as free cholesterol), cholesterol ester, total fatty acid. , Phosphoric lipids, fatty acids (eg, oleic acid, etc.), fatty alcohols and the like. The total of cholesterol ester and free cholesterol is total cholesterol (also referred to as TC (total cholesterol)).
後述の試験例でも示すように、本発明は、ジクアホソルがＰ２Ｙ２受容体シグナル伝達を介して、マイボーム腺細胞から全分泌型で脂質分泌を促進するという結果に基づく。よって、本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物は、脂質分泌促進によって予防または治療される眼疾患、脂質の補充を必要とする眼疾患、脂質減少に起因する眼疾患などの予防または治療に有用である。例えば、マイボーム腺機能不全（ＭＧＤ）、眼瞼炎、ドライアイ（脂質減少に起因するドライアイを含む）などが挙げられるが、これらの眼疾患に限定されるものではない。
As shown in Test Examples below, the present invention is, diquafosol via the P2Y 2 receptor signaling, based on the result of promoting lipid secretion in all secreted from meibomian gland cells. Therefore, the lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention are caused by eye diseases prevented or treated by promoting lipid secretion, eye diseases requiring lipid supplementation, and lipid reduction. It is useful for the prevention or treatment of eye diseases. Examples include, but are not limited to, meibomian gland dysfunction (MGD), blepharitis, dry eye (including dry eye due to lipid loss), and the like.
ドライアイは「様々な要因による涙液および角結膜上皮の慢性疾患であり、眼不快感や視覚異常を伴う疾患」と定義づけられ、乾性角結膜炎（ＫＣＳ：ｋｅｒａｔｏｃｏｎｊｕｎｃｔｉｖｉｔｉｓ ｓｉｃｃａ）はドライアイに含まれる。本発明においては、ソフトコンタクトレンズ装用を原因とするドライアイ症状の発生もドライアイに含まれるものとする。
Dry eye is defined as "a chronic disease of tears and keratoconjunctival epithelium caused by various factors and is accompanied by eye discomfort and visual abnormalities", and keratoconjunctivitis sicca (KCS) is included in dry eye. .. In the present invention, the occurrence of dry eye symptoms caused by wearing soft contact lenses is also included in dry eye.
ドライアイ症状には、眼乾燥感、眼不快感、眼疲労感、鈍重感、羞明感、眼痛、霧視（かすみ目）などの自覚症状の他、充血、角結膜上皮障害などの他覚所見も含まれる。
Dry eye symptoms include subjective symptoms such as dry eyes, discomfort, eye fatigue, dullness, dullness, eye pain, and blurred vision (blurred vision), as well as other symptoms such as congestion and keratoconjunctival epithelial disorders. Findings are also included.
ドライアイの病因については不明点も多いが、シェーグレン症候群；先天性無涙腺症；サルコイドーシス；骨髄移植による移植片対宿主病（ＧＶＨＤ：Ｇｒａｆｔ Ｖｅｒｓｕｓ Ｈｏｓｔ Ｄｉｓｅａｓｅ）；眼類天疱瘡；スティーブンス・ジョンソン症候群；トラコーマなどを原因とする涙器閉塞；糖尿病；角膜屈折矯正手術（ＬＡＳＩＫ：Ｌａｓｅｒ（－ａｓｓｉｓｔｅｄ） ｉｎ Ｓｉｔｕ Ｋｅｒａｔｏｍｉｌｅｕｓｉｓ）などを原因とする反射性分泌の低下；マイボーム腺機能不全；涙液層の不安定化（安定性低下）；涙液減少；脂質（脂質層）の減少；眼瞼炎などを原因とする油層（脂質層）の減少；眼球突出、兎眼などを原因とする瞬目不全または閉瞼不全；杯細胞からのムチン分泌低下；ＶＤＴ（Ｖｉｓｕａｌ Ｄｉｓｐｌａｙ Ｔｅｒｍｉｎａｌｓ）作業などがその原因／起因であると報告されている。
There are many unclear points about the etiology of dry eye, but Sjogren's syndrome; congenital tear adenopathy; sarcoidosis; transplant-to-host disease by bone marrow transplantation (GVHD: Graft Versus Host Disease); ocular pyorrhea; Stevens Johnson syndrome Lacrimal obstruction caused by tracoma, etc .; Diabetes; Decreased reflex secretion caused by LASIK (LASIK: -assisted) in Situ Keratomilisis; Mybohm gland dysfunction; Tear layer failure Stabilization (decreased stability); Decreased tears; Decreased lipid (lipid layer); Decreased oil layer (lipid layer) caused by eyelid inflammation, etc .; It has been reported that the cause / cause is eyelid insufficiency; decreased mutin secretion from cup cells; VDT (Visual Display Thermals) work and the like.
マイボーム腺機能不全（ＭＧＤ）の定義は、例えば、「さまざまな原因によってマイボーム腺の機能が瀰漫性に異常をきたした状態であり、慢性の眼不快感を伴う」というものである。ここで、「マイボーム腺の機能が瀰漫性に異常をきたした状態」とは、例えば、霰粒腫、内麦粒腫などで認められる局所的なマイボーム腺異常ではなく、毛細血管拡張、マイボーム腺開口部閉塞などのマイボーム腺異常が瀰漫性に認められることを意味する。また、ＭＧＤは、分泌減少型ＭＧＤ（ｌｏｗ－ｄｅｌｉｖｅｒｙ ｓｔａｔｅ）と分泌増加型ＭＧＤ（ｈｉｇｈ ｄｅｌｉｖｅｒｙ ｓｔａｔｅ）に分けられ、更に分泌減少型ＭＧＤ（ｌｏｗ－ｄｅｌｉｖｅｒｙ ｓｔａｔｅ ＭＤ）として、「ｈｙｐｏｓｅｃｒｅｔｏｒｙ ＭＧＤ（ｍｅｉｂｏｍｉａｎ ｈｙｐｏｓｅｃｒｅｔｉｏｎ）」および「ｏｂｓｔｒｕｃｔｉｖｅ ＭＧＤ（ｍｅｉｂｏｍｉａｎ ｇｌａｎｄ ｏｂｓｔｒｕｃｔｉｏｎ）」が挙げられる。
The definition of meibomian gland dysfunction (MGD) is, for example, "a state in which the function of the meibomian glands is diffusely abnormal due to various causes, accompanied by chronic eye discomfort." Here, the "state in which the function of the meibomian gland is diffusely abnormal" is not a local meibomian gland abnormality observed in, for example, chalazion or hordeolum, but is a dilation of the meibomian gland or obstruction of the opening of the meibomian gland. It means that meibomian gland abnormalities such as are found diffusely. Further, the MGD is divided into a secretion-reducing MGD (low-delivery state) and a secretion-increasing MGD (high-delivery state), and further as a secretion-reducing MGD (low-delivery state MD), "hyposectory MGD (meibomian gland) meibomian gland". "And" meibomian gland obstruction ".
分泌減少型ＭＧＤでは、マイボーム腺開口部の閉塞などによってマイボーム腺脂の分泌が減少する。また、分泌増加型ＭＧＤでは、種々の原因によってマイボーム腺脂の分泌が増加する。
In the reduced secretion type MGD, the secretion of meibomian gland fat is reduced due to obstruction of the meibomian gland opening. In addition, in the secretory-increasing MGD, the secretion of meibomian gland fat increases due to various causes.
一方、マイボーム腺開口部の閉塞が認められるものの、自覚症状を伴わない状態を、例えば、「マイボーム腺梗塞（ｍｅｉｂｏｍｉａｎ ｇｌａｎｄ ｃｏｎｃｒｅｔｉｏｎ）」と呼ぶこともあるが、本発明におけるＭＧＤには、マイボーム腺梗塞も含まれるものとする。ジクアホソルが治療しうるＭＧＤとして特に好ましいのは、分泌減少型ＭＧＤである。
On the other hand, a state in which obstruction of the meibomian gland opening is observed but not accompanied by subjective symptoms is sometimes referred to as, for example, "meibomian gland concretion", but the MGD in the present invention refers to meibomian gland infarction. Is also included. Particularly preferred as the MGD that diquafosol can treat is a hyposecretory MGD.
ところで、ＭＧＤはドライアイの原因となることもあり、また、後部眼瞼炎を引き起こす可能性もある。ＭＧＤには、「ドライアイおよび／または後部眼瞼炎を伴う（合併する）ＭＧＤ」、「ドライアイおよび／または後部眼瞼炎の原因となるＭＧＤ」、「ドライアイを伴わない（合併しない）ＭＧＤ」、「ドライアイの原因とならないＭＧＤ」、「後部眼瞼炎を伴わない（合併しない）ＭＧＤ」ならびに「後部眼瞼炎の原因とならないＭＧＤ」が含まれる。眼瞼炎とは、瞼が炎症を起こしている症状であり、前部眼瞼炎、眼瞼縁炎、後部眼瞼炎、眼瞼皮膚炎、眼角眼瞼炎などが含まれる。
By the way, MGD may cause dry eye and may also cause posterior blepharitis. MGD includes "MGD with (combined) dry eye and / or posterior blepharitis", "MGD causing dry eye and / or posterior blepharitis", "MGD without (uncomplicated) dry eye". , "MGD that does not cause dry eye", "MGD that does not accompany (do not combine) with posterior blepharitis" and "MGD that does not cause posterior blepharitis" are included. Blepharitis is a symptom of inflamed eyelids and includes anterior blepharitis, blepharitis, posterior blepharitis, blepharitis, and blepharitis.
本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物は、インビボ投与に使用することができる。また、本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物は、インビボ処理方法に使用することができる。
The lipid secretion promoter, tear layer stabilizer, and ophthalmic composition of the present invention can be used for in vivo administration. In addition, the lipid secretion promoter, tear layer stabilizer, and ophthalmic composition of the present invention can be used in an in vivo treatment method.
本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物は、眼疾患などの予防および／または治療に使用することが好ましい。
The lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention are preferably used for the prevention and / or treatment of eye diseases and the like.
本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物の剤型としては、例えば点眼剤、眼軟膏、注射剤、軟膏（例えば、眼瞼皮膚に投与できる）などが挙げられ、好ましくは点眼剤である。ここで点眼剤とは点眼液または点眼薬と同義であり、コンタクトレンズ用点眼剤も点眼剤の定義に含まれる。
Examples of the dosage form of the lipid secretion promoter, the tear film stabilizer, and the ophthalmic composition of the present invention include eye drops, eye ointments, injections, ointments (for example, which can be administered to the eyelid skin) and the like. It is preferably an eye drop. Here, eye drops are synonymous with eye drops or eye drops, and eye drops for contact lenses are also included in the definition of eye drops.
本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物は、水を溶媒（基剤）とすることが好ましく、水性点眼剤であることがより好ましい。
The lipid secretion promoter, tear layer stabilizer, and ophthalmic composition of the present invention preferably use water as a solvent (base), and more preferably an aqueous eye drop.
本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物は、有効成分や添加物の性質、含量などによって、溶解型点眼剤であってもよく、懸濁型点眼剤であってもよい。
The lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention may be a soluble eye drop or a suspension type eye drop depending on the properties and contents of the active ingredient and the additive. May be.
本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物には、汎用されている技術を用い、必要に応じて製薬学的に許容される添加剤をさらに添加することができる。例えば、リン酸ナトリウム、リン酸水素ナトリウム、リン酸二水素ナトリウム、酢酸ナトリウム、イプシロン－アミノカプロン酸などの緩衝化剤；塩化カルシウム、塩化ナトリウム、塩化カリウム、濃グリセリンなどの等張化剤；エデト酸ナトリウムなどの安定化剤；ポリソルベートなどの界面活性剤；アスコルビン酸などの抗酸化剤；塩化ベンザルコニウム、クロルヘキシジングルコン酸塩などの防腐剤；塩酸、水酸化ナトリウムなどのｐＨ調節剤などを必要に応じて選択し、添加することができる。これらの添加剤は、１種単独で使用してもよく、また２種以上を任意に組み合わせて使用してもよい。
The lipid secretion promoter, tear film stabilizer, and ophthalmic composition of the present invention can be further added with pharmaceutically acceptable additives as needed by using a general-purpose technique. .. For example, buffering agents such as sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium acetate, epsilon-aminocaproic acid; isotonic agents such as calcium chloride, sodium chloride, potassium chloride, concentrated glycerin; edetonic acid. Stabilizers such as sodium; Surface active agents such as polysorbate; Antioxidants such as ascorbic acid; Preservatives such as benzalkonium chloride and chlorhexizing luconate; pH adjusters such as hydrochloric acid and sodium hydroxide are required. It can be selected and added accordingly. These additives may be used alone or in any combination of two or more.
本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物のｐＨは、医薬として許容される範囲であれば特定の値に限定されない。しかしながら、本発明の脂質分泌促進剤、涙液層安定化剤、眼科用組成物のｐＨは、好ましくは８以下、より好ましくは４～８の範囲、さらに好ましくは５～８の範囲、よりさらに好ましくは６～８の範囲、特に好ましくは７の近傍である。
The pH of the lipid secretion promoter, the tear film stabilizer, and the ophthalmic composition of the present invention is not limited to a specific value as long as it is within the range acceptable for pharmaceuticals. However, the pH of the lipid secretion promoter, the tear film stabilizer, and the ophthalmic composition of the present invention is preferably 8 or less, more preferably in the range of 4 to 8, still more preferably in the range of 5 to 8, and even more. It is preferably in the range of 6 to 8, particularly preferably in the vicinity of 7.
本発明はまた、マイボーム腺細胞からの全分泌型の脂質分泌を促進するためのインビボ処理方法において使用するための、ジクアホソルまたはその塩を含有する眼科用組成物についても提供する。
The present invention also provides an ophthalmic composition containing diquafosol or a salt thereof for use in an in vivo treatment method for promoting holocrine lipid secretion from meibomian gland cells.
本発明はさらに、マイボーム腺細胞からの脂質分泌を促進するためのインビボ処理方法において使用するための、ジクアホソルまたはその塩を含有する眼科用組成物であって、該脂質がワックスエステル、トリグリセリド、オメガ水酸化脂質、コレステロール、コレステロールエステル、総コレステロール、リン脂質、脂肪酸および脂肪アルコールからなる群から選択される少なくとも１つである、眼科用組成物についても提供する。
The present invention further comprises an ophthalmic composition comprising diquafosol or a salt thereof for use in an in vivo treatment method for promoting lipid secretion from mybome gland cells, wherein the lipid is a wax ester, triglyceride, omega water. Also provided is an ophthalmic composition which is at least one selected from the group consisting of lipid oxides, cholesterol, cholesterol esters, total cholesterol, phospholipids, fatty acids and fatty alcohols.
以下に、薬理試験の結果および製剤例を示すが、これらの例は本発明をよりよく理解するためのものであり、本発明の範囲を限定するものではない。
The results of pharmacological tests and pharmaceutical examples are shown below, but these examples are for better understanding of the present invention and do not limit the scope of the present invention.
［試験例１］
ジクアホソルがマイボーム腺細胞における細胞内カルシウムイオンに与える影響を検討した。 [Test Example 1]
The effect of diquafosol on intracellular calcium ions in meibomian gland cells was investigated.
ジクアホソルがマイボーム腺細胞における細胞内カルシウムイオンに与える影響を検討した。 [Test Example 1]
The effect of diquafosol on intracellular calcium ions in meibomian gland cells was investigated.
（試料調製）
ジクアホソルナトリウムを精製水に溶解して、８．５％（ｗ／ｖ）水溶液を調製した後、Ｈａｍ Ｆ－１２で希釈して目的濃度のジクアホソルナトリウム（以下、「ＤＱＳ」と略）液を調製した。 (Sample preparation)
Diquafosol sodium is dissolved in purified water to prepare an 8.5% (w / v) aqueous solution, which is then diluted with Ham F-12 to the desired concentration of diquafosol sodium (hereinafter abbreviated as "DQS"). ) Liquid was prepared.
ジクアホソルナトリウムを精製水に溶解して、８．５％（ｗ／ｖ）水溶液を調製した後、Ｈａｍ Ｆ－１２で希釈して目的濃度のジクアホソルナトリウム（以下、「ＤＱＳ」と略）液を調製した。 (Sample preparation)
Diquafosol sodium is dissolved in purified water to prepare an 8.5% (w / v) aqueous solution, which is then diluted with Ham F-12 to the desired concentration of diquafosol sodium (hereinafter abbreviated as "DQS"). ) Liquid was prepared.
（試験方法）
白色家兎より酵素処理にてマイボーム腺細胞を単離し、各種培養添加物を含むＨａｍ Ｆ－１２培地中で無血清培養した。使用数日前に細胞分化を誘導し、細胞内カルシウム指示蛍光色素を含む緩衝液（Ｓｃｒｅｅｎ Ｑｕｅｓｔ Ｃａｌｉｂｒｙｔｅ５２０ Ｃａｌｃｉｕｍ Ａｓｓａｙ Ｋｉｔ、ＡＡＴ Ｂｉｏｑｕｅｓｔ社）中でインキュベーションした。室温下で静置し、その間にＰ２Ｙ２受容体アンタゴニスト適用群にはＡＲ－Ｃ１１８９２５ＸＸ（以下、「ＡＲ－Ｃ」と略、終濃度：３μＭ）を加えた。蛍光プレートリーダーを用いてカルシウム指示色素の経時的蛍光測定を開始した。開始から６０秒後に緩衝液９容に対して１容のＤＱＳ液（終濃度：０．０１５％、０．０４％、０．１％、０．３％、０．８５％）を添加し、蛍光強度の測定を継続した。 (Test method)
Meibomian gland cells were isolated from white rabbits by enzyme treatment and cultured without serum in Ham F-12 medium containing various culture additives. Cell differentiation was induced several days before use and incubated in a buffer containing an intracellular calcium-indicating fluorescent dye (Screen Quest Calibrite 520 Assay Kit, AAT Bioquest). Standing at room temperature, during which the P2Y 2 receptor antagonists applied group AR-C118925XX (hereinafter generally as "AR-C", final concentration: 3 [mu] M) was added. The fluorescence measurement of the calcium indicator dye over time was started using a fluorescent plate reader. 60 seconds after the start, 1 volume of DQS solution (final concentration: 0.015%, 0.04%, 0.1%, 0.3%, 0.85%) was added to 9 volumes of buffer solution. The measurement of fluorescence intensity was continued.
白色家兎より酵素処理にてマイボーム腺細胞を単離し、各種培養添加物を含むＨａｍ Ｆ－１２培地中で無血清培養した。使用数日前に細胞分化を誘導し、細胞内カルシウム指示蛍光色素を含む緩衝液（Ｓｃｒｅｅｎ Ｑｕｅｓｔ Ｃａｌｉｂｒｙｔｅ５２０ Ｃａｌｃｉｕｍ Ａｓｓａｙ Ｋｉｔ、ＡＡＴ Ｂｉｏｑｕｅｓｔ社）中でインキュベーションした。室温下で静置し、その間にＰ２Ｙ２受容体アンタゴニスト適用群にはＡＲ－Ｃ１１８９２５ＸＸ（以下、「ＡＲ－Ｃ」と略、終濃度：３μＭ）を加えた。蛍光プレートリーダーを用いてカルシウム指示色素の経時的蛍光測定を開始した。開始から６０秒後に緩衝液９容に対して１容のＤＱＳ液（終濃度：０．０１５％、０．０４％、０．１％、０．３％、０．８５％）を添加し、蛍光強度の測定を継続した。 (Test method)
Meibomian gland cells were isolated from white rabbits by enzyme treatment and cultured without serum in Ham F-12 medium containing various culture additives. Cell differentiation was induced several days before use and incubated in a buffer containing an intracellular calcium-indicating fluorescent dye (Screen Quest Calibrite 520 Assay Kit, AAT Bioquest). Standing at room temperature, during which the P2Y 2 receptor antagonists applied group AR-C118925XX (hereinafter generally as "AR-C", final concentration: 3 [mu] M) was added. The fluorescence measurement of the calcium indicator dye over time was started using a fluorescent plate reader. 60 seconds after the start, 1 volume of DQS solution (final concentration: 0.015%, 0.04%, 0.1%, 0.3%, 0.85%) was added to 9 volumes of buffer solution. The measurement of fluorescence intensity was continued.
（評価方法）
ＤＱＳ液添加直前の蛍光強度（ＲＦＵ：ｒｅｌａｔｉｖｅ ｆｌｕｏｒｅｓｃｅｎｃｅ ｕｎｉｔｓ）を基準に、添加後に最大蛍光強度を示した時点での蛍光強度増加率（ＲＦＵ ｉｎｃｒｅａｓｅ（％ ｏｆ ｂａｓｅｌｉｎｅ））を算出した。 (Evaluation method)
Based on the fluorescence intensity (RFU) immediately before the addition of the DQS solution, the fluorescence intensity increase rate (RFU increase (% of baseline)) at the time when the maximum fluorescence intensity was shown after the addition was calculated.
ＤＱＳ液添加直前の蛍光強度（ＲＦＵ：ｒｅｌａｔｉｖｅ ｆｌｕｏｒｅｓｃｅｎｃｅ ｕｎｉｔｓ）を基準に、添加後に最大蛍光強度を示した時点での蛍光強度増加率（ＲＦＵ ｉｎｃｒｅａｓｅ（％ ｏｆ ｂａｓｅｌｉｎｅ））を算出した。 (Evaluation method)
Based on the fluorescence intensity (RFU) immediately before the addition of the DQS solution, the fluorescence intensity increase rate (RFU increase (% of baseline)) at the time when the maximum fluorescence intensity was shown after the addition was calculated.
（結果）
結果を図１に示す。図１に示すように、ジクアホソルの添加後、細胞内のカルシウムイオンのシグナル伝達は劇的に増加した。また、その増加は濃度依存的であった。一方、Ｐ２Ｙ２受容体のアンタゴニストであるＡＲ－Ｃの共存下では、その増加はほぼ完全に抑制された。 (result)
The results are shown in FIG. As shown in FIG. 1, intracellular calcium ion signaling increased dramatically after the addition of diquafosol. Moreover, the increase was concentration-dependent. On the other hand, in the presence of AR-C is an antagonist of P2Y 2 receptors, the increase was almost completely inhibited.
結果を図１に示す。図１に示すように、ジクアホソルの添加後、細胞内のカルシウムイオンのシグナル伝達は劇的に増加した。また、その増加は濃度依存的であった。一方、Ｐ２Ｙ２受容体のアンタゴニストであるＡＲ－Ｃの共存下では、その増加はほぼ完全に抑制された。 (result)
The results are shown in FIG. As shown in FIG. 1, intracellular calcium ion signaling increased dramatically after the addition of diquafosol. Moreover, the increase was concentration-dependent. On the other hand, in the presence of AR-C is an antagonist of P2Y 2 receptors, the increase was almost completely inhibited.
（考察）
上記の結果から、ジクアホソルがターゲットとするプリン作動性Ｐ２Ｙ２受容体のシグナル伝達機構がマイボーム腺細胞に存在することが示された。 (Discussion)
From the above results, diquafosol signal transduction mechanism of purinergic P2Y 2 receptor targeting has been shown to be present in meibomian gland cells.
上記の結果から、ジクアホソルがターゲットとするプリン作動性Ｐ２Ｙ２受容体のシグナル伝達機構がマイボーム腺細胞に存在することが示された。 (Discussion)
From the above results, diquafosol signal transduction mechanism of purinergic P2Y 2 receptor targeting has been shown to be present in meibomian gland cells.
［試験例２］
ジクアホソルがマイボーム腺細胞からの脂質分泌に与える影響を検討した。 [Test Example 2]
We investigated the effect of diquafosol on lipid secretion from meibomian gland cells.
ジクアホソルがマイボーム腺細胞からの脂質分泌に与える影響を検討した。 [Test Example 2]
We investigated the effect of diquafosol on lipid secretion from meibomian gland cells.
（試料調製）
ジクアホソルナトリウムを精製水に溶解して、８．５％（ｗ／ｖ）水溶液を調製した後、Ｈａｍ Ｆ－１２で希釈して目的濃度のジクアホソルナトリウム（以下、「ＤＱＳ」と略）液を調製した。 (Sample preparation)
Diquafosol sodium is dissolved in purified water to prepare an 8.5% (w / v) aqueous solution, which is then diluted with Ham F-12 to the desired concentration of diquafosol sodium (hereinafter abbreviated as "DQS"). ) Liquid was prepared.
ジクアホソルナトリウムを精製水に溶解して、８．５％（ｗ／ｖ）水溶液を調製した後、Ｈａｍ Ｆ－１２で希釈して目的濃度のジクアホソルナトリウム（以下、「ＤＱＳ」と略）液を調製した。 (Sample preparation)
Diquafosol sodium is dissolved in purified water to prepare an 8.5% (w / v) aqueous solution, which is then diluted with Ham F-12 to the desired concentration of diquafosol sodium (hereinafter abbreviated as "DQS"). ) Liquid was prepared.
（試験方法）
白色家兎より酵素処理にてマイボーム腺細胞を単離し、各種培養添加物を含むＨａｍ Ｆ－１２培地中で無血清培養した。細胞分化を誘導した後、細胞を洗浄し、各種濃度のジクアホソルナトリウム（終濃度：０．０３％、０．３％、０．８５％）を添加した。なおＰ２Ｙ２受容体アンタゴニストを適用する群にはＡＲ－Ｃ１１８９２５ＸＸ（以下、「ＡＲ－Ｃ」と略、終濃度：３μＭ）も同時に添加した。４時間インキュベーションした後、培養上清と細胞とを別々に回収した。培養上清サンプルはそのまま、他方、細胞サンプルは脂質成分を抽出処理した後に、それぞれ総コレステロール定量試薬（Ａｍｐｒｅｘ Ｒｅｄ Ｃｈｏｌｅｓｔｅｒｏｌ Ａｓｓａｙ、Ｉｎｖｉｔｒｏｇｅｎ社）を用いて総コレステロール（ＴＣ）含量を測定した。 (Test method)
Meibomian gland cells were isolated from white rabbits by enzyme treatment and cultured without serum in Ham F-12 medium containing various culture additives. After inducing cell differentiation, the cells were washed and various concentrations of diquafosol sodium (final concentrations: 0.03%, 0.3%, 0.85%) were added. Note in the group applying the P2Y 2 receptor antagonist AR-C118925XX (hereinafter generally as "AR-C", final concentration: 3 [mu] M) was also added at the same time. After incubation for 4 hours, the culture supernatant and cells were collected separately. The culture supernatant sample was left as it was, while the cell sample was subjected to extraction treatment of lipid components, and then the total cholesterol (TC) content was measured using a total cholesterol quantifying reagent (Amplex Red Cholesterol Assay, Invitrogen).
白色家兎より酵素処理にてマイボーム腺細胞を単離し、各種培養添加物を含むＨａｍ Ｆ－１２培地中で無血清培養した。細胞分化を誘導した後、細胞を洗浄し、各種濃度のジクアホソルナトリウム（終濃度：０．０３％、０．３％、０．８５％）を添加した。なおＰ２Ｙ２受容体アンタゴニストを適用する群にはＡＲ－Ｃ１１８９２５ＸＸ（以下、「ＡＲ－Ｃ」と略、終濃度：３μＭ）も同時に添加した。４時間インキュベーションした後、培養上清と細胞とを別々に回収した。培養上清サンプルはそのまま、他方、細胞サンプルは脂質成分を抽出処理した後に、それぞれ総コレステロール定量試薬（Ａｍｐｒｅｘ Ｒｅｄ Ｃｈｏｌｅｓｔｅｒｏｌ Ａｓｓａｙ、Ｉｎｖｉｔｒｏｇｅｎ社）を用いて総コレステロール（ＴＣ）含量を測定した。 (Test method)
Meibomian gland cells were isolated from white rabbits by enzyme treatment and cultured without serum in Ham F-12 medium containing various culture additives. After inducing cell differentiation, the cells were washed and various concentrations of diquafosol sodium (final concentrations: 0.03%, 0.3%, 0.85%) were added. Note in the group applying the P2Y 2 receptor antagonist AR-C118925XX (hereinafter generally as "AR-C", final concentration: 3 [mu] M) was also added at the same time. After incubation for 4 hours, the culture supernatant and cells were collected separately. The culture supernatant sample was left as it was, while the cell sample was subjected to extraction treatment of lipid components, and then the total cholesterol (TC) content was measured using a total cholesterol quantifying reagent (Amplex Red Cholesterol Assay, Invitrogen).
（評価方法）
培養上清中のＴＣ含量と細胞内ＴＣ含量の総和に対する培養上清中に存在するＴＣの百分率（Ｒａｔｉｏ ｏｆ ｒｅｌｅａｓｅｄ ＴＣ）（％）を算出し、脂質分泌度の指標とした。 (Evaluation method)
The percentage of TC present in the culture supernatant (Ratio of relaxed TC) (%) with respect to the sum of the TC content in the culture supernatant and the intracellular TC content was calculated and used as an index of the degree of lipid secretion.
培養上清中のＴＣ含量と細胞内ＴＣ含量の総和に対する培養上清中に存在するＴＣの百分率（Ｒａｔｉｏ ｏｆ ｒｅｌｅａｓｅｄ ＴＣ）（％）を算出し、脂質分泌度の指標とした。 (Evaluation method)
The percentage of TC present in the culture supernatant (Ratio of relaxed TC) (%) with respect to the sum of the TC content in the culture supernatant and the intracellular TC content was calculated and used as an index of the degree of lipid secretion.
（結果）
結果を図２に示す。図２に示すように、マイボーム腺細胞から一定の割合でＴＣが細胞外（培養上清中）へと放出され、その放出されたＴＣ比率はＤＱＳの濃度依存的に増加した。一方、Ｐ２Ｙ２受容体のアンタゴニストであるＡＲ－Ｃの共存下ではＤＱＳによるＴＣ分泌の増加は顕著に抑制された。 (result)
The results are shown in FIG. As shown in FIG. 2, TC was released extracellularly (in the culture supernatant) from the meibomian gland cells at a constant rate, and the released TC ratio increased in a DQS concentration-dependent manner. On the other hand, the increase in TC secretion by DQS is in the presence of AR-C is an antagonist of P2Y 2 receptor was significantly suppressed.
結果を図２に示す。図２に示すように、マイボーム腺細胞から一定の割合でＴＣが細胞外（培養上清中）へと放出され、その放出されたＴＣ比率はＤＱＳの濃度依存的に増加した。一方、Ｐ２Ｙ２受容体のアンタゴニストであるＡＲ－Ｃの共存下ではＤＱＳによるＴＣ分泌の増加は顕著に抑制された。 (result)
The results are shown in FIG. As shown in FIG. 2, TC was released extracellularly (in the culture supernatant) from the meibomian gland cells at a constant rate, and the released TC ratio increased in a DQS concentration-dependent manner. On the other hand, the increase in TC secretion by DQS is in the presence of AR-C is an antagonist of P2Y 2 receptor was significantly suppressed.
（考察）
以上の結果より、ジクアホソルがＰ２Ｙ２受容体シグナル伝達を介して、マイボーム腺細胞における脂質分泌を促進することが示された。 (Discussion)
These results, diquafosol via the P2Y 2 receptor signaling was shown to promote lipid secretion in meibomian gland cells.
以上の結果より、ジクアホソルがＰ２Ｙ２受容体シグナル伝達を介して、マイボーム腺細胞における脂質分泌を促進することが示された。 (Discussion)
These results, diquafosol via the P2Y 2 receptor signaling was shown to promote lipid secretion in meibomian gland cells.
［試験例３］
ジクアホソルで処理したマイボーム腺細胞におけるアポトーシスの発現について検討した。 [Test Example 3]
The expression of apoptosis in meibomian gland cells treated with diquafosol was investigated.
ジクアホソルで処理したマイボーム腺細胞におけるアポトーシスの発現について検討した。 [Test Example 3]
The expression of apoptosis in meibomian gland cells treated with diquafosol was investigated.
（試料調製）
ジクアホソルナトリウムを精製水に溶解して、８．５％（ｗ／ｖ）水溶液を調製し、培養液９容に対して本ジクアホソルナトリウム（以下、「ＤＱＳ」と略）液１容の割合で添加使用した。 (Sample preparation)
Dissolve diquafosol sodium in purified water to prepare an 8.5% (w / v) aqueous solution, and 1 volume of this diquafosol sodium (hereinafter abbreviated as "DQS") solution for 9 volumes of the culture solution. Was added and used in the ratio of.
ジクアホソルナトリウムを精製水に溶解して、８．５％（ｗ／ｖ）水溶液を調製し、培養液９容に対して本ジクアホソルナトリウム（以下、「ＤＱＳ」と略）液１容の割合で添加使用した。 (Sample preparation)
Dissolve diquafosol sodium in purified water to prepare an 8.5% (w / v) aqueous solution, and 1 volume of this diquafosol sodium (hereinafter abbreviated as "DQS") solution for 9 volumes of the culture solution. Was added and used in the ratio of.
（試験方法）
白色家兎より酵素処理にてマイボーム腺細胞を単離し、各種培養添加物を含むＨａｍ Ｆ－１２培地中で無血清培養した。細胞分化を誘導した後、洗浄し、ＤＱＳ液を添加して２時間～６時間インキュベーションした。培養上清とともに細胞を回収し、ＡｐｏｐＬａｄｄｅｒ Ｅｘ（タカラバイオ社）を用いて各々の断片化ＤＮＡサンプルを得た。１．５％ アガロースゲルで断片化ＤＮＡサンプルを電気泳動した後、ＳＹＢＲ Ｇｒｅｅｎ Ｉ色素でゲルを染色し、ＤＮＡのバンドを可視化した。また同時に、断片化ＤＮＡサンプルにＳＹＢＲ Ｇｒｅｅｎ Ｉ色素を加えて蛍光強度を測定し、断片化ＤＮＡ量を定量した。 (Test method)
Meibomian gland cells were isolated from white rabbits by enzyme treatment and cultured without serum in Ham F-12 medium containing various culture additives. After inducing cell differentiation, the cells were washed, DQS solution was added, and the cells were incubated for 2 to 6 hours. The cells were collected together with the culture supernatant, and each fragmented DNA sample was obtained using ApopRadder Ex (Takara Bio Inc.). Fragmented DNA samples were electrophoresed on a 1.5% agarose gel and then stained with SYBR Green I dye to visualize DNA bands. At the same time, SYBR Green I dye was added to the fragmented DNA sample, the fluorescence intensity was measured, and the amount of fragmented DNA was quantified.
白色家兎より酵素処理にてマイボーム腺細胞を単離し、各種培養添加物を含むＨａｍ Ｆ－１２培地中で無血清培養した。細胞分化を誘導した後、洗浄し、ＤＱＳ液を添加して２時間～６時間インキュベーションした。培養上清とともに細胞を回収し、ＡｐｏｐＬａｄｄｅｒ Ｅｘ（タカラバイオ社）を用いて各々の断片化ＤＮＡサンプルを得た。１．５％ アガロースゲルで断片化ＤＮＡサンプルを電気泳動した後、ＳＹＢＲ Ｇｒｅｅｎ Ｉ色素でゲルを染色し、ＤＮＡのバンドを可視化した。また同時に、断片化ＤＮＡサンプルにＳＹＢＲ Ｇｒｅｅｎ Ｉ色素を加えて蛍光強度を測定し、断片化ＤＮＡ量を定量した。 (Test method)
Meibomian gland cells were isolated from white rabbits by enzyme treatment and cultured without serum in Ham F-12 medium containing various culture additives. After inducing cell differentiation, the cells were washed, DQS solution was added, and the cells were incubated for 2 to 6 hours. The cells were collected together with the culture supernatant, and each fragmented DNA sample was obtained using ApopRadder Ex (Takara Bio Inc.). Fragmented DNA samples were electrophoresed on a 1.5% agarose gel and then stained with SYBR Green I dye to visualize DNA bands. At the same time, SYBR Green I dye was added to the fragmented DNA sample, the fluorescence intensity was measured, and the amount of fragmented DNA was quantified.
（結果）
結果を図３に示す。図３（ａ）に示すように、培養したマイボーム腺細胞から調製した断片化ＤＮＡより、アポトーシス細胞に特徴的な梯子状のＤＮＡバンドが検出された。またこの特徴的なＤＮＡのラダーパターンはＤＱＳ添加後も乱れることはなかった。一方、定量的には、図３（ｂ）に示すように、断片化ＤＮＡ量はＤＱＳ添加４時間後に大きく増加した。すなわち、マイボーム腺細胞で構成的に発生しているアポトーシス性細胞死がＤＱＳ添加後に促進されることが示された。 (result)
The results are shown in FIG. As shown in FIG. 3A, a ladder-shaped DNA band characteristic of apoptotic cells was detected in the fragmented DNA prepared from cultured meibomian gland cells. Moreover, this characteristic DNA ladder pattern was not disturbed even after the addition of DQS. On the other hand, quantitatively, as shown in FIG. 3 (b), the amount of fragmented DNA increased significantly 4 hours after the addition of DQS. That is, it was shown that the apoptotic cell death constitutively occurring in meibomian gland cells is promoted after the addition of DQS.
結果を図３に示す。図３（ａ）に示すように、培養したマイボーム腺細胞から調製した断片化ＤＮＡより、アポトーシス細胞に特徴的な梯子状のＤＮＡバンドが検出された。またこの特徴的なＤＮＡのラダーパターンはＤＱＳ添加後も乱れることはなかった。一方、定量的には、図３（ｂ）に示すように、断片化ＤＮＡ量はＤＱＳ添加４時間後に大きく増加した。すなわち、マイボーム腺細胞で構成的に発生しているアポトーシス性細胞死がＤＱＳ添加後に促進されることが示された。 (result)
The results are shown in FIG. As shown in FIG. 3A, a ladder-shaped DNA band characteristic of apoptotic cells was detected in the fragmented DNA prepared from cultured meibomian gland cells. Moreover, this characteristic DNA ladder pattern was not disturbed even after the addition of DQS. On the other hand, quantitatively, as shown in FIG. 3 (b), the amount of fragmented DNA increased significantly 4 hours after the addition of DQS. That is, it was shown that the apoptotic cell death constitutively occurring in meibomian gland cells is promoted after the addition of DQS.
（考察）
以上の結果より、ジクアホソルはマイボーム腺細胞の自発的な細胞死を促進することが示された。試験例２の結果と考え併せると、ジクアホソルはマイボーム腺細胞の全分泌型脂質分泌を促進することが示された。 (Discussion)
From the above results, it was shown that diquafosol promotes spontaneous cell death of meibomian gland cells. Combined with the results of Test Example 2, it was shown that diquafosol promotes holocrine lipid secretion in meibomian gland cells.
以上の結果より、ジクアホソルはマイボーム腺細胞の自発的な細胞死を促進することが示された。試験例２の結果と考え併せると、ジクアホソルはマイボーム腺細胞の全分泌型脂質分泌を促進することが示された。 (Discussion)
From the above results, it was shown that diquafosol promotes spontaneous cell death of meibomian gland cells. Combined with the results of Test Example 2, it was shown that diquafosol promotes holocrine lipid secretion in meibomian gland cells.
［製剤例］
製剤例を挙げて本発明の薬剤をさらに具体的に説明するが、本発明はこれらの製剤例にのみ限定されるものではない。 [Example of pharmaceutical product]
The agent of the present invention will be described in more detail with reference to pharmaceutical examples, but the present invention is not limited to these pharmaceutical examples.
製剤例を挙げて本発明の薬剤をさらに具体的に説明するが、本発明はこれらの製剤例にのみ限定されるものではない。 [Example of pharmaceutical product]
The agent of the present invention will be described in more detail with reference to pharmaceutical examples, but the present invention is not limited to these pharmaceutical examples.
滅菌精製水にジクアホソルナトリウムおよびそれ以外の成分を加え、これらを十分に混合することで点眼剤などの各製剤を調製できる。
By adding diquafosol sodium and other components to sterilized purified water and mixing them sufficiently, each formulation such as eye drops can be prepared.
（処方例１：点眼剤（３％（ｗ／ｖ）））
１００ｍｌ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例２：点眼剤（３％（ｗ／ｖ）））
１００ｍｌ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
ＢＡＫ－Ｃ１２ ０．１～１０ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例３：点眼剤（３％（ｗ／ｖ）））
１００ｍｌ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
ＢＡＫ－Ｃ１２ ０．１～１０ｇ
クエン酸一水和物 ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例４：点眼剤（３％（ｗ／ｖ）））
１００ｍｌ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
ＢＡＫ－Ｃ１２ ０．１～１０ｇ
メタリン酸ナトリウム ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例５：点眼剤（３％（ｗ／ｖ）））
１００ｍｌ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
ＢＡＫ－Ｃ１２ ０．１～１０ｇ
ポリリン酸ナトリウム ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例６：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
ポリビニルピロリドン Ｋ９０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例７：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
ポリビニルピロリドン Ｋ６０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例８：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
ポリビニルピロリドン Ｋ４０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例９：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
ポリビニルピロリドン Ｋ９０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１０：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
ポリビニルピロリドン Ｋ６０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１１：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
ポリビニルピロリドン Ｋ４０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１２：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
クエン酸水和物 ０．０００１～０．１ｇ
ポリビニルピロリドン Ｋ９０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１３：点眼剤（３％（ｗ／ｖ）））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
クロルヘキシジングルコン酸塩 ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１４：点眼剤（３％（ｗ／ｖ）））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
クロルヘキシジングルコン酸塩 ０．０００１～０．１ｇ
ポリソルベート８０ ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１５：点眼剤（３％（ｗ／ｖ）））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
塩化ベンザルコニウム ０．０００１～１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量 (Prescription example 1: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100ml
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 2: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100ml
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
BAK-C12 0.1-10g
Sodium edetate hydrate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 3: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100ml
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
BAK-C12 0.1-10g
Citric acid monohydrate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 4: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100ml
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
BAK-C12 0.1-10g
Sodium metaphosphate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 5: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100ml
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
BAK-C12 0.1-10g
Sodium polyphosphate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 6: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Polyvinylpyrrolidone K90 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 7: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Polyvinylpyrrolidone K60 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription Example 8: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Polyvinylpyrrolidone K40 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 9: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Polyvinylpyrrolidone K90 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 10: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Polyvinylpyrrolidone K60 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 11: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Polyvinylpyrrolidone K40 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 12: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Citric acid hydrate 0.0001-0.1g
Polyvinylpyrrolidone K90 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 13: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Chlorhexidine gluconate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 14: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Chlorhexidine gluconate 0.0001-0.1g
Polysorbate 80 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 15: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
Benzalkonium chloride 0.0001 to 1 g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount
１００ｍｌ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例２：点眼剤（３％（ｗ／ｖ）））
１００ｍｌ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
ＢＡＫ－Ｃ１２ ０．１～１０ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例３：点眼剤（３％（ｗ／ｖ）））
１００ｍｌ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
ＢＡＫ－Ｃ１２ ０．１～１０ｇ
クエン酸一水和物 ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例４：点眼剤（３％（ｗ／ｖ）））
１００ｍｌ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
ＢＡＫ－Ｃ１２ ０．１～１０ｇ
メタリン酸ナトリウム ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例５：点眼剤（３％（ｗ／ｖ）））
１００ｍｌ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
ＢＡＫ－Ｃ１２ ０．１～１０ｇ
ポリリン酸ナトリウム ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例６：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
ポリビニルピロリドン Ｋ９０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例７：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
ポリビニルピロリドン Ｋ６０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例８：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
ポリビニルピロリドン Ｋ４０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例９：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
ポリビニルピロリドン Ｋ９０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１０：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
ポリビニルピロリドン Ｋ６０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１１：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
ポリビニルピロリドン Ｋ４０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１２：点眼剤（３％（ｗ／ｖ））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．０１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
クエン酸水和物 ０．０００１～０．１ｇ
ポリビニルピロリドン Ｋ９０ ０．０００１～１０ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１３：点眼剤（３％（ｗ／ｖ）））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
クロルヘキシジングルコン酸塩 ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１４：点眼剤（３％（ｗ／ｖ）））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
エデト酸ナトリウム水和物 ０．０００１～０．１ｇ
クロルヘキシジングルコン酸塩 ０．０００１～０．１ｇ
ポリソルベート８０ ０．０００１～０．１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量
（処方例１５：点眼剤（３％（ｗ／ｖ）））
１００ｍＬ中
ジクアホソルナトリウム ３ｇ
リン酸水素ナトリウム水和物 ０．１～０．５ｇ
塩化ナトリウム ０．０１～１ｇ
塩化カリウム ０．０１～１ｇ
塩化ベンザルコニウム ０．０００１～１ｇ
ｐＨ調節剤 適量
滅菌精製水 適量 (Prescription example 1: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100ml
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 2: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100ml
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
BAK-C12 0.1-10g
Sodium edetate hydrate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 3: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100ml
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
BAK-C12 0.1-10g
Citric acid monohydrate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 4: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100ml
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
BAK-C12 0.1-10g
Sodium metaphosphate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 5: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100ml
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
BAK-C12 0.1-10g
Sodium polyphosphate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 6: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Polyvinylpyrrolidone K90 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 7: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Polyvinylpyrrolidone K60 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription Example 8: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Polyvinylpyrrolidone K40 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 9: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Polyvinylpyrrolidone K90 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 10: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Polyvinylpyrrolidone K60 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 11: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Polyvinylpyrrolidone K40 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 12: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium Hydrogen Phosphate Hydrate 0.01-0.5g
Sodium chloride 0.01-1g
Citric acid hydrate 0.0001-0.1g
Polyvinylpyrrolidone K90 0.0001-10g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 13: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Chlorhexidine gluconate 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 14: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
Sodium edetate hydrate 0.0001-0.1g
Chlorhexidine gluconate 0.0001-0.1g
Polysorbate 80 0.0001-0.1g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount (Prescription example 15: Eye drops (3% (w / v)))
Diquafosol sodium 3g in 100mL
Sodium hydrogen phosphate hydrate 0.1-0.5g
Sodium chloride 0.01-1g
Potassium chloride 0.01-1g
Benzalkonium chloride 0.0001 to 1 g
Acidity regulator Appropriate amount Sterilized purified water Appropriate amount
ジクアホソルまたはその塩はＰ２Ｙ２受容体シグナル伝達を介して、マイボーム腺細胞における脂質分泌を増強することから優れた脂質分泌促進作用を有し、脂質分泌促進によって予防または治療される眼疾患、脂質の補充を必要とする眼疾患、脂質減少に起因する眼疾患等の予防または治療に有用である。
Diquafosol or a salt thereof via the P2Y 2 receptor signaling, has excellent lipid secretion promoting action because it enhances lipid secretion in meibomian gland cells, ocular disease to be prevented or treated by lipid secretagogue, recruitment of lipids It is useful for the prevention or treatment of eye diseases that require meibomian glands, eye diseases caused by lipid reduction, and the like.
Claims (7)
- ジクアホソルまたはその塩を有効成分として含有する脂質分泌促進剤であって、脂質分泌が全分泌型である、脂質分泌促進剤。 A lipid secretion promoter containing diquafosol or a salt thereof as an active ingredient, which is a holocrine type of lipid secretion.
- 脂質分泌がマイボーム腺細胞からの分泌である、請求項１に記載の脂質分泌促進剤。 The lipid secretion promoter according to claim 1, wherein the lipid secretion is secretion from meibomian gland cells.
- 脂質が、ワックスエステル、トリグリセリド、オメガ水酸化脂質、コレステロール、コレステロールエステル、総コレステロール、リン脂質、脂肪酸および脂肪アルコールからなる群から選択される少なくとも１つである、請求項１または２に記載の脂質分泌促進剤。 The lipid according to claim 1 or 2, wherein the lipid is at least one selected from the group consisting of wax esters, triglycerides, omega hydroxide lipids, cholesterol, cholesterol esters, total cholesterol, phospholipids, fatty acids and fatty alcohols. Secretion promoter.
- 請求項１～３のいずれか１項に記載の脂質分泌促進剤を含有する、涙液層安定化剤。 A tear layer stabilizer containing the lipid secretion promoter according to any one of claims 1 to 3.
- 脂質分泌を促進するための、ジクアホソルまたはその塩を有効成分として含有する眼科用組成物であって、脂質分泌が全分泌型である、眼科用組成物。 An ophthalmic composition containing diquafosol or a salt thereof as an active ingredient for promoting lipid secretion, wherein the lipid secretion is a holocrine type.
- マイボーム腺細胞からの全分泌型の脂質分泌を促進するためのインビボ処理方法において使用するための、ジクアホソルまたはその塩を含有する眼科用組成物。 An ophthalmic composition containing diquafosol or a salt thereof for use in an in vivo treatment method for promoting holocrine lipid secretion from meibomian gland cells.
- マイボーム腺細胞からの脂質分泌を促進するためのインビボ処理方法において使用するための、ジクアホソルまたはその塩を含有する眼科用組成物であって、該脂質がワックスエステル、トリグリセリド、オメガ水酸化脂質、コレステロール、コレステロールエステル、総コレステロール、リン脂質、脂肪酸および脂肪アルコールからなる群から選択される少なくとも１つである、眼科用組成物。 An ophthalmic composition containing diquafosol or a salt thereof for use in an in vivo treatment method for promoting lipid secretion from mybome gland cells, wherein the lipid is a wax ester, triglyceride, omega hydroxide lipid, cholesterol, An ophthalmic composition which is at least one selected from the group consisting of cholesterol esters, total cholesterol, phospholipids, fatty acids and fatty alcohols.
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| JP2020-110332 | 2020-06-26 | ||
| JP2020110332A JP2023133646A (en) | 2020-06-26 | 2020-06-26 | Lipid secretagogue |
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| JP (1) | JP2023133646A (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2011527709A (en) * | 2008-07-10 | 2011-11-04 | インスパイアー ファーマシューティカルズ，インコーポレイティド | How to treat blepharitis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2011527709A (en) * | 2008-07-10 | 2011-11-04 | インスパイアー ファーマシューティカルズ，インコーポレイティド | How to treat blepharitis |
Non-Patent Citations (4)
| Title |
|---|
| ENDO KEN-ICHI, SAKAMOTO ASUKA, FUJISAWA KOUSHI: "Diquafosol tetrasodium elicits total cholesterol release from rabbit meibomian gland cells via P2Y2 purinergic receptor signalling", SCIENTIFIC REPORTS, vol. 11, no. 1, 26 March 2021 (2021-03-26), pages 6989, XP055893672, DOI: 10.1038/s41598-021-86433-6 * |
| FUKUOKA SHIMA, ARITA REIKO: "Increase in tear film lipid layer thickness after instillation of 3% diquafosol ophthalmic solution in healthy human eyes", OCULAR SURFACE, vol. 15, no. 4, 1 October 2017 (2017-10-01), pages 730 - 735, XP055893665, ISSN: 1542-0124, DOI: 10.1016/j.jtos.2017.03.005 * |
| FUKUOKA SHIMA, ARITA REIKO: "Tear film lipid layer increase after diquafosol instillation in dry eye patients with meibomian gland dysfunction: a randomized clinical study", SCIENTIFIC REPORTS, vol. 9, no. 9091, 24 June 2019 (2019-06-24), XP055893668, DOI: 10.1038/s41598-019-45475-7 * |
| SIN MAN LAM, LOUIS TONG, SIEW SIAN YONG, BOWEN LI, SHYAM S. CHAURASIA, GUANGHOU SHUI, MARKUS R. WENK: "Meibum Lipid Composition in Asians with Dry Eye Disease", PLOS ONE, PUBLIC LIBRARY OF SCIENCE, vol. 6, no. 10, 1 January 2011 (2011-01-01), pages e24339, XP055013727, ISSN: 19326203, DOI: 10.1371/journal.pone.0024339 * |
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