WO2015152416A1 - Ocular hypotensive agent - Google Patents

Ocular hypotensive agent Download PDF

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WO2015152416A1
WO2015152416A1 PCT/JP2015/060676 JP2015060676W WO2015152416A1 WO 2015152416 A1 WO2015152416 A1 WO 2015152416A1 JP 2015060676 W JP2015060676 W JP 2015060676W WO 2015152416 A1 WO2015152416 A1 WO 2015152416A1
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intraocular pressure
tie2
activator
lowering agent
schlemm
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PCT/JP2015/060676
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French (fr)
Japanese (ja)
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和一 丸山
健太朗 加治屋
美加 加治屋
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国立大学法人東北大学
株式会社資生堂
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Publication of WO2015152416A1 publication Critical patent/WO2015152416A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
    • A61K36/254—Acanthopanax or Eleutherococcus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/54—Lauraceae (Laurel family), e.g. cinnamon or sassafras
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • 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
    • 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
    • A61P27/06—Antiglaucoma agents or miotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention provides an intraocular pressure-lowering agent comprising a Tie2 activating (phosphorylating) agent.
  • Glaucoma In Japan, an aging society, eye diseases associated with aging are increasing. At present, glaucoma is the leading cause of blindness, and it is estimated that there are 4 million patients in Japan including potential glaucoma patients. Glaucoma affects approximately 5% of people over the age of 40 and is currently the leading cause of blindness in Japan. Glaucoma is an optic neuropathy that causes RGC thinning due to the impairment of retinal ganglion cells (RGC) that control visual function, resulting in visual field impairment.
  • RGC retinal ganglion cells
  • One of the main causes of RGC disorders is increased intraocular pressure.
  • the only mainstream of glaucoma treatment is to slow the visual field progression with the only evidenced treatment for reducing intraocular pressure.
  • the central method of glaucoma treatment is to reduce intraocular pressure using eye drops or internal medicine.
  • the hemodynamics of the eyeball are also important, and it has been reported that disturbance of ocular blood circulation causes glaucoma to progress. In any case, the development of further new preventive methods and treatment methods suitable for the disease state is desired.
  • aqueous humor is responsible for supplying nutrients.
  • Aqueous humor is produced in the ciliary body, and about 90% or more flows out to the venous system outside the eye via Schlemm's canal. Since the anatomical study of Schlemm's canal itself has not been sufficiently conducted, it has not yet been clarified through what mechanism of Schlemm's canal the drainage of aqueous humor.
  • Angiopoietin (angiopoietin; Ang) is known as a factor involved in vascular structure stabilization and angiogenesis.
  • Ang is known to activate the receptor tyrosine kinase Tie (tyrosineroskinase with Ig and EGF homology domain) -2 expressed in vascular endothelial cells and exert its function (Non-patent Document 1).
  • Tie2 activators such as Ang regulate adhesion between vascular endothelial cells and vascular wall cells such as pericytes (pericytes) and vascular smooth muscle cells via Tie-2 expressed in vascular endothelial cells. It is understood that it functions in the structural stabilization of blood vessels (Non-patent Document 2).
  • a lymphatic vessel stabilizer comprising a Tie2 (tyrosine kinase with Ig and EGF homology domain 2) activator
  • the Tie2 activator comprises Angiopoietin 1 (Ang-1), Nikkei (Cinnamomum ) Genus plant extract, Siberian carrot (SiberianiberGinseng) extract, syringaresinol and the like are also disclosed.
  • Ang-1 Angiopoietin 1
  • Nikkei Cinnamomum
  • Siberian carrot SiberianiberGinseng
  • An object of the present invention is to provide an intraocular pressure-lowering agent comprising a Tie2 activating (phosphorylating) agent.
  • aqueous humor a type of fluid produced and secreted by the ciliary body.
  • About 90% of the aqueous humor has been elucidated until it is discharged through Schlemm's canal, but histoanatomical knowledge of Schlemm's canal itself has not been obtained, and the outflow mechanism of the aqueous humor has not been elucidated.
  • the present inventor examined the physiological and biochemical properties of Schlemm's canal and found that Tie2 was expressed in Schlemm's canal, like lymphatics. This finding is a completely new discovery and may be a marker for new Schlemm's canal endothelium.
  • a Tie2 activator decreases intraocular pressure as a result of research, and has completed the following invention:
  • (1-1) An intraocular pressure-lowering agent comprising a Tie2 (tyrosine kinase with Ig and EGF homology domain 2) activator.
  • 1-2 The intraocular pressure-lowering agent according to (1-1), wherein the Tie2 activator activates Tie2 in Schlemm's canal.
  • 1-3 The intraocular pressure-lowering agent according to (1-1) or (1-2), wherein the Tie2 activator is an extract of Siberian Ginseng and / or Keihi.
  • (1-5) A composition comprising the intraocular pressure-lowering agent according to any one of (1-1) to (1-4).
  • (1-6) A pharmaceutical composition comprising the intraocular pressure-lowering agent according to any one of (1-1) to (1-4).
  • (1-7) The pharmaceutical composition according to (1-6) for preventing and / or treating glaucoma.
  • the pharmaceutical composition according to (1-6) or (1-7) which is in a dosage form for oral administration.
  • (3-1) A method for reducing intraocular pressure by administering a Tie2 activator to a subject who requires a reduction in intraocular pressure.
  • (3-2) The method according to (3-1), wherein the Tie2 activator activates Tie2 in Schlemm's canal.
  • (3-3) The method according to (3-1) or (3-2), wherein the Tie2 activator is an extract of Siberian ginseng and / or Keihi.
  • (3-4) The method according to any one of (3-1) to (3-3), wherein the Tie2 activator is an alcohol extract of Siberian ginseng and / or Keihi.
  • administration of the Tie2 activator can activate Tie2 in Schlemm's canal and reduce intraocular pressure.
  • a food composition such as a pharmaceutical composition or a supplement containing an intraocular pressure-lowering agent comprising a Tie2 activator can be used for preventing and / or treating glaucoma.
  • FIG. 1 shows the expression state of Tie2 in Schlemm's canal.
  • FIG. 1A shows an anatomical schematic diagram of the eye.
  • FIG. 1B shows a HE-stained ocular tissue section (4 ⁇ ).
  • FIG. 1C shows an enlargement of FIG. 1B (10 times), and the arrow points to Schlemm's canal.
  • FIG. 1D shows the same tissue stained with anti-Tie2 antibody, with arrows pointing to Schlemm's canal stained with anti-Tie2 antibody.
  • FIG. 1E shows nuclear staining of the same tissue with Hoechst, with arrows pointing to Schlemm's canal.
  • FIG. 2 shows intraocular pressure at 9:00 and 17:00 when the Siberian carrot extract was orally administered at 9:00 am.
  • FIG. 3 shows intraocular pressure at each time when the Siberian carrot extract was orally administered at 8 am.
  • FIG. 4 shows intraocular pressure at 9:00 and 17:00 when the cinnamon extract is orally administered at 9:00 am.
  • the present invention relates to an intraocular pressure-lowering agent comprising a Tie2 activator.
  • the present invention also relates to food compositions such as pharmaceutical compositions and supplements containing an intraocular pressure-lowering agent comprising a Tie2 activator. Since the food composition such as the pharmaceutical composition or supplement of the present invention has an extremely excellent intraocular pressure-lowering action, it can be used for the prevention and / or treatment of glaucoma and ocular hypertension.
  • Conventionally known intraocular pressure-lowering agents include sympathomimetic drugs, sympathetic nerve blockers, parasympathomimetic drugs, prostaglandin-related drugs, carbonic acid dehydrogenase inhibitors, and the like. In the past, no intraocular pressure-lowering agent that promotes proper drainage of aqueous humor by activating Tie2 in Schlemm's canal is known.
  • the activator of Tie2 is not particularly limited, but is known to have an activity to activate Tie2, such as Angiopoietin 1, or an extract derived from a plant of the genus Cinnamomum, Siberian carrot (SiberianiberGinseng) ) Extracts or syringalesinol, eleutheroside E (Eleutheroside), eleutheroside E1, and the like that the present inventors have found to have the activity.
  • the activation of Tie2 here refers to the ability to phosphorylate Tie2 and convert it into its active form (phosphorylated Tie2).
  • Nikkei is a plant of the order of Lauraceae and Lauraceae, and there are more than 300 species, such as Cinnamomum cassia Blume, C. camphora, C. daphnoides (C. doederleinii), C. japonicum, Ogasawara jay nick (C. pseudo-pedunculatum), Nikkei (C. zeylanicum) is known.
  • an extract derived from Cay (Cinnamomum cassia Blume), in particular, Cay nae (Keishi) or Keihi (Cinnamon), which is a young branch of Cay, is used.
  • Siberian carrot (Siberian Ginseng) is also called Ezoukogi and is a plant belonging to the family Argiaceae.
  • the medicinal part is the root bark, which has long been used as a raw material for traditional Chinese medicine and herbal medicines.
  • the above-mentioned extract can be obtained by a conventional method, for example, it can be obtained by soaking or heating and refluxing the plant that is the origin together with the extraction solvent at room temperature or heating, followed by filtration and concentration.
  • the extraction solvent can be arbitrarily used as long as it is a solvent that is usually used for extraction.
  • an aqueous solvent such as water, physiological saline, phosphate buffer, borate buffer, or an organic solvent such as ethanol, Alcohols such as propylene glycol, 1,3-butylene glycol and glycerin, hydrous alcohols, chloroform, dichloroethane, carbon tetrachloride, acetone, ethyl acetate, hexane and the like can be used alone or in combination.
  • water is used as the solvent.
  • the extract obtained by extraction with the above solvent can be used as it is or, for example, an extract concentrated by lyophilization or the like, and if necessary, an adsorbent method, for example, an ion exchange resin removed impurities, After adsorption on a polymer column (for example, Amberlite XAD-2), elution with a desired solvent, and further concentrated or only active ingredients can be used.
  • an adsorbent method for example, an ion exchange resin removed impurities
  • the dosage form of the pharmaceutical composition of the present invention may be oral, parenteral, external use and the like.
  • Examples of the dosage form include oral administration agents such as tablets, powders, capsules, granules, extracts and syrups, or parenteral ointments such as injections, drops, eye drops, or suppositories, creams, Mention may be made of external preparations such as emulsions and lotions.
  • Examples of the dosage form of the food composition include supplements, granules, drinks and the like.
  • the blending amount of the Tie2 activator in the pharmaceutical composition or food composition of the present invention can be appropriately determined depending on the use, but is generally 0.0001 to 20.0 mol%, preferably 0.0001 to 10.0 mol% in the total amount of the agent. It is.
  • compositions and food composition of the present invention for example, excipients, moisture-proofing agents, preservatives, reinforcing agents, thickeners, emulsifiers, antioxidants used in ordinary foods and pharmaceuticals.
  • sweeteners, acidulants, seasonings, colorants, fragrances and the like can be appropriately blended as necessary.
  • the ophthalmic pharmaceutical composition of the present invention can be used in combination with a conventional pharmaceutically acceptable carrier depending on the dosage form.
  • the ophthalmic pharmaceutical composition of the present invention may optionally contain various components such as carbohydrates, electrolytes, amino acids, vitamins, lipids, pharmaceutical additives, and pharmaceuticals.
  • components include, for example, sugars such as glucose, maltose, oligosaccharides, mannitol, sugar alcohols such as sorbitol; electrolytes such as sodium chloride, sodium hydrogen phosphate, potassium chloride, magnesium sulfate, chloride Calcium, etc .; amino acids such as glycine, alanine, etc .; vitamins such as thiamine hydrochloride, sodium riboflavin phosphate, pyridoxine hydrochloride, nicotinamide, folic acid, biotin, vitamin A, L-ascorbic acid, ⁇ -glycosyl ascorbic acid and the like These derivatives may be mentioned, and these may be combined in combination as necessary.
  • ophthalmic pharmaceutical composition of the present invention examples include commonly used additives such as preservatives such as methyl parahydroxybenzoate, sodium dehydroacetate, benzalkonium chloride and the like; stabilizers such as sodium edetate and sodium bisulfite. Buffers such as borax, boric acid, sodium hydrogen carbonate, etc .; thickeners such as methyl cellulose, carboxymethyl cellulose, chondroitin sulfate, polyvinyl alcohol, pullulan, etc .; solubilizers such as polysorbate 80 may be blended.
  • preservatives such as methyl parahydroxybenzoate, sodium dehydroacetate, benzalkonium chloride and the like
  • stabilizers such as sodium edetate and sodium bisulfite. Buffers such as borax, boric acid, sodium hydrogen carbonate, etc .
  • thickeners such as methyl cellulose, carboxymethyl cellulose, chondroitin sulfate, polyvinyl alcohol, pullulan, etc
  • a cholinergic intraocular pressure-lowering agent e.g., pilocarpine, carbachol excellent in miosis effect
  • anticholinesterase e.g., deme potassium, DFP, ecothiol
  • physostigmine salicylate as a miotic agent
  • pilocarpine hydrochloride mannitol, glycerin, isosorbide, etc.
  • FIG. 1C is an enlarged view of a HE-stained eyeball tissue section, and the arrow points to Schlemm's canal.
  • FIG. 1D shows the same tissue stained with anti-Tie2 antibody, and the arrow indicates Schlemm's canal stained with anti-Tie2 antibody. The staining of Schlemm's canal indicates that Tie2 is expressed in Schlemm's canal.
  • Experiment 2 Intraocular pressure measurement test 1 (Siberian carrot) The subject had an intraocular pressure of 22 mmHg or less with a Goldman applanation tonometer, and no ocular abnormalities were observed in slit lamp microscopy and fundus examination. ⁇ Health history, history of systemic diseases such as hypertension and diabetes, smoking history, and healthy adults with no eye drops or internal history.
  • Siberian ginseng powder a Siberian carrot extract obtained by extracting 30% ethanol from Siberian carrot roots (Ask Chemical), was used. Expect a sample size of 20 (10 men and women) and administer Siberian ginseng powder to all subjects.
  • the wash-out period is longer than 1 month, and three internal doses and before and after intraocular pressure, blood pressure, pulse, and laser speckle tests are performed.
  • the primary endpoint is a decrease in intraocular pressure
  • the secondary endpoint is an increase in ocular blood flow as measured by laser speckle flowography.
  • Statistical analysis was performed on Siberian ginseng powder at these endpoints.
  • FIG. 2 shows intraocular pressure at the time of ingestion (9 am) and 17:00. After ingestion of Siberian ginseng powder, a decrease in intraocular pressure was observed in about 3 hours, which lasted for about 8 hours.
  • the control was the same subject who did not take Siberian ginseng powder, and the intraocular pressure was measured on a different day from the day of taking Siberian ginseng powder.
  • Experiment 3 Intraocular pressure measurement test 2 (Siberian carrot) The same conditions as in Experiment 2 were used except that the internal use was 8:00 am and the intraocular pressure was measured with a non-contact tonometer.
  • FIG. 3 shows intraocular pressure at each time. After taking Siberian ginseng powder, the intraocular pressure decreased until about 10 hours.
  • Experiment 4 Intraocular pressure measurement test (Keihi) The subject had an intraocular pressure of 22 mmHg or less with a Goldman applanation tonometer, and no ocular abnormalities were observed in slit lamp microscopy and fundus examination. ⁇ Health history, history of systemic diseases such as hypertension and diabetes, smoking history, and healthy adults with no eye drops or internal history.
  • Keihi extract powder obtained by extracting Keihi bark with 30% ethanol was used.
  • the sample size is 8 (5 women and 3 men), and 100 mg of keihi extract powder is ingested at 9 am for all subjects (with commercial mineral water: product name;
  • the intraocular pressure was measured with a Goldman applanation tonometer.
  • the wash-out period was 1 month or longer, and a single oral dose and before and after intraocular pressure, blood pressure, pulse, and laser speckle tests were performed.
  • the primary endpoint is a decrease in intraocular pressure
  • the secondary endpoint is an increase in ocular blood flow as measured by laser speckle flowography.
  • Statistical analysis (ANOVA) was conducted on Keihi powder for these endpoints.
  • FIG. 4 shows the intraocular pressure of the left eye at each time.

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Abstract

The purpose of the present invention is to provide a novel ocular hypotensive agent. The ocular hypotensive agent comprises a Tie2 (tyrosine kinase with Ig and EGF homology domain 2) activator.

Description

眼圧降下剤Intraocular pressure-lowering agent
　本発明はTie2活性化（リン酸化）剤からなる、眼圧降下剤を提供する。 The present invention provides an intraocular pressure-lowering agent comprising a Tie2 activating (phosphorylating) agent.
　高齢化社会となった日本では、加齢に伴う眼疾患が増加している。現在、緑内障は中途失明の原因の第一位となり、潜在的な緑内障患者を含めるとわが国には４００万人の患者がいることが推測されている。緑内障は40歳以上の約5%が罹患し、現在本邦において失明原因第一位の疾患である。緑内障は、視機能を司る網膜神経節細胞(Retinal ganglion cell; RGC)の障害に伴いRGCの菲薄化を生じ、その結果として視野障害を生じる視神経症である。RGCの障害の主な原因の一つは眼圧の上昇である。現状では唯一エビデンスのある眼圧下降治療により視野進行を遅らせることが緑内障治療の主流となっている。緑内障治療の中心となる方法は点眼薬や内服薬を用いて眼圧を低下させることである。しかしながら、緑内障において眼圧の低下に加え眼球の血行動態もまた重要であり、眼球血流循環の障害が緑内障を進行させることが報告されている。いずれにせよ病態に即した更なる新しい予防法や治療法の開発が望まれている。 In Japan, an aging society, eye diseases associated with aging are increasing. At present, glaucoma is the leading cause of blindness, and it is estimated that there are 4 million patients in Japan including potential glaucoma patients. Glaucoma affects approximately 5% of people over the age of 40 and is currently the leading cause of blindness in Japan. Glaucoma is an optic neuropathy that causes RGC thinning due to the impairment of retinal ganglion cells (RGC) that control visual function, resulting in visual field impairment. One of the main causes of RGC disorders is increased intraocular pressure. At present, the only mainstream of glaucoma treatment is to slow the visual field progression with the only evidenced treatment for reducing intraocular pressure. The central method of glaucoma treatment is to reduce intraocular pressure using eye drops or internal medicine. However, in addition to the decrease in intraocular pressure in glaucoma, the hemodynamics of the eyeball are also important, and it has been reported that disturbance of ocular blood circulation causes glaucoma to progress. In any case, the development of further new preventive methods and treatment methods suitable for the disease state is desired.
　眼圧は、房水と呼ばれる眼内を循環する体液の一種により制御されている。眼には透過性の高い血管（脳血流関門のような構造であり、透過性が低い）及びリンパ管が存在しないため、房水が栄養の供給を担っている。房水は毛様体で産生され、約９０％以上がシュレム管を経由して眼外の静脈系へと流出する。シュレム管自体の組織解剖学的な研究は十分にされていないため、シュレム管のどのような機構を通じて房水の排出が行われているのかは未だ解明されていない。 O Intraocular pressure is controlled by a kind of bodily fluid circulating in the eye called aqueous humor. Since the eye has no highly permeable blood vessels (structure like a cerebral blood flow barrier and low permeability) and lymphatic vessels, aqueous humor is responsible for supplying nutrients. Aqueous humor is produced in the ciliary body, and about 90% or more flows out to the venous system outside the eye via Schlemm's canal. Since the anatomical study of Schlemm's canal itself has not been sufficiently conducted, it has not yet been clarified through what mechanism of Schlemm's canal the drainage of aqueous humor.
　血管構造の安定化や血管新生に関与する因子としてアンジオポエチン（angiopoietin；Ang）が知られる。Angは血管内皮細胞に発現する受容体型チロシンキナーゼTie（tyrosine kinase with Ig and EGF homology domain）-2を活性化させ、その機能を発揮することで知られる（非特許文献１）。Ang等のTie2活性化剤は、血管内皮細胞に発現するTie-2を介し、血管内皮細胞と、周皮細胞（ペリサイト）や血管平滑筋細胞のような血管壁細胞との接着を制御し、血管の構造的安定化に機能していることが理解されている（非特許文献２）。また、Tie2（tyrosine kinase with Ig and EGF homology domain 2）活性化剤からなる、リンパ管の安定化剤が開示されており、該Tie2活性化剤が、アンジオポエチン１(Ang-1)、ニッケイ（Cinnamomum)属植物の抽出物、シベリアニンジン（Siberian Ginseng）抽出物及びシリンガレシノールなどであることも開示されている。しかしながら、上記のとおり、そもそも眼内には透過性の高い血管もリンパ管も存在しないため、Tie-2と眼圧との関係が知られる由もない。 Angiopoietin (angiopoietin; Ang) is known as a factor involved in vascular structure stabilization and angiogenesis. Ang is known to activate the receptor tyrosine kinase Tie (tyrosineroskinase with Ig and EGF homology domain) -2 expressed in vascular endothelial cells and exert its function (Non-patent Document 1). Tie2 activators such as Ang regulate adhesion between vascular endothelial cells and vascular wall cells such as pericytes (pericytes) and vascular smooth muscle cells via Tie-2 expressed in vascular endothelial cells. It is understood that it functions in the structural stabilization of blood vessels (Non-patent Document 2). In addition, a lymphatic vessel stabilizer comprising a Tie2 (tyrosine kinase with Ig and EGF homology domain 2) activator is disclosed, and the Tie2 activator comprises Angiopoietin 1 (Ang-1), Nikkei (Cinnamomum ) Genus plant extract, Siberian carrot (SiberianiberGinseng) extract, syringaresinol and the like are also disclosed. However, as described above, there are no highly permeable blood vessels or lymphatic vessels in the eye, so there is no reason to know the relationship between Tie-2 and intraocular pressure.
国際公開第2009/154237号International Publication No. 2009/154237
　本発明の課題は、Tie2活性化（リン酸化）剤からなる、眼圧降下剤の提供にある。 An object of the present invention is to provide an intraocular pressure-lowering agent comprising a Tie2 activating (phosphorylating) agent.
　冒頭で述べたとおり、眼内圧は毛様体より産生・分泌される体液の一種である房水により調節される。房水の約90％はシュレム管を通じて排出されることまでは解明されているが、シュレム管自体の組織解剖学的な知見は得られておらず、房水流出機構は解明されていない。本発明者は、シュレム管の生理学的・生化学的諸性質を調べたところ、シュレム管にはリンパ管と同様、Tie2が発現していることを見出した。この知見は全くの新規発見であり、また新規シュレム管内皮のマーカーになる可能性がある。この知見に基づき、Tie2活性化剤を投与してみたところ、コントロールと比較して眼圧が低下することを発見した。特に理論に拘束されるものではないが、これは、Tie2活性化剤の投与により、シュレム管におけるTie2が活性化され、房水が適切に排出されることによると考えられる。 As described at the beginning, intraocular pressure is regulated by aqueous humor, a type of fluid produced and secreted by the ciliary body. About 90% of the aqueous humor has been elucidated until it is discharged through Schlemm's canal, but histoanatomical knowledge of Schlemm's canal itself has not been obtained, and the outflow mechanism of the aqueous humor has not been elucidated. The present inventor examined the physiological and biochemical properties of Schlemm's canal and found that Tie2 was expressed in Schlemm's canal, like lymphatics. This finding is a completely new discovery and may be a marker for new Schlemm's canal endothelium. Based on this finding, administration of a Tie2 activator found that intraocular pressure was reduced compared to controls. Although not particularly bound by theory, it is thought that this is due to the activation of Tie2 in Schlemm's canal by administration of the Tie2 activator and proper drainage of aqueous humor.
　発明者らは、研究の結果、Tie2活性化剤の投与が、眼圧を低下させることを見出し、以下の発明を完成するに至った：
　(1-1)Tie2（tyrosine kinase with Ig and EGF homology domain2）活性化剤からなる、眼圧降下剤。
　(1-2)前記Tie2活性化剤が、シュレム管におけるTie2を活性化する(1-1)に記載の眼圧降下剤。
　(1-3)前記Tie2活性化剤が、シベリアニンジン（Siberian Ginseng）及び／又はケイヒの抽出物である、(1-1)又は(1-2)に記載の眼圧降下剤。
　(1-4)前記Tie2活性化剤が、シベリアニンジン（Siberian Ginseng）及び／又はケイヒのアルコール抽出物である、(1-1)～(1-3)のいずれかに記載の眼圧降下剤。
　(1-5)(1-1)～(1-4)のいずれかに記載の眼圧降下剤を含有する、組成物。
　(1-6)(1-1)～(1-4)のいずれかに記載の眼圧降下剤を含有する、医薬組成物。
　(1-7)緑内障を予防及び／又は治療するための、(1-6)に記載の医薬組成物。
　(1-8)経口投与の剤型にある、(1-6)又は(1-7)に記載の医薬組成物。
　(1-9)点眼剤の剤型にある、(1-6)又は(1-7)に記載の医薬組成物。
　(2-1)眼圧低下のために使用するための、Tie2活性化剤。
　(2-2)シュレム管におけるTie2を活性化する、(2-1)に記載のTie2活性化剤。
　(2-3)シベリアニンジン及び／又はケイヒの抽出物である、(2-1)又は(2-2)に記載のTie2活性化剤。
　(2-4)シベリアニンジン及び／又はケイヒのアルコール抽出物である、(2-1)～(2-3)のいずれかに記載のTie2活性化剤。
　(2-5)緑内障を予防及び／又は治療するための、(2-1)～(2-4)のいずれかに記載のTie2活性化剤。
　(2-6)前記Tie2活性化剤が、経口で投与される、(2-1)～(2-5)のいずれかに記載のTie2活性化剤。
　(2-7)前記Tie2活性化剤が、点眼剤として投与される、(2-1)～(2-5)のいずれかに記載のTie2活性化剤。
　(3-1)眼圧低下を必要とする対象者にTie2活性化剤を投与することによる、眼圧低下方法。
　(3-2)前記Tie2活性化剤が、シュレム管におけるTie2を活性化する、(3-1)に記載の方法。
　(3-3)前記Tie2活性化剤が、シベリアニンジン及び／又はケイヒの抽出物である、(3-1)又は(3-2)に記載の方法。
　(3-4)前記Tie2活性化剤が、シベリアニンジン及び／又はケイヒのアルコール抽出物である、(3-1)～(3-3)のいずれかに記載の方法。
　(3-5)緑内障を予防及び／又は治療するための、(3-1)～(3-4)のいずれかに記載の方法。
　(3-6)前記Tie2活性化剤が、経口で投与される、(3-1)～(3-5)のいずれかに記載の方法。
　(3-7)前記Tie2活性化剤が、点眼剤として投与される、(3-1)～(3-5)のいずれかに記載の方法。 The inventors have found that administration of a Tie2 activator decreases intraocular pressure as a result of research, and has completed the following invention:
(1-1) An intraocular pressure-lowering agent comprising a Tie2 (tyrosine kinase with Ig and EGF homology domain 2) activator.
(1-2) The intraocular pressure-lowering agent according to (1-1), wherein the Tie2 activator activates Tie2 in Schlemm's canal.
(1-3) The intraocular pressure-lowering agent according to (1-1) or (1-2), wherein the Tie2 activator is an extract of Siberian Ginseng and / or Keihi.
(1-4) The intraocular pressure-lowering agent according to any one of (1-1) to (1-3), wherein the Tie2 activator is an alcohol extract of Siberian Ginseng and / or Keihi .
(1-5) A composition comprising the intraocular pressure-lowering agent according to any one of (1-1) to (1-4).
(1-6) A pharmaceutical composition comprising the intraocular pressure-lowering agent according to any one of (1-1) to (1-4).
(1-7) The pharmaceutical composition according to (1-6) for preventing and / or treating glaucoma.
(1-8) The pharmaceutical composition according to (1-6) or (1-7), which is in a dosage form for oral administration.
(1-9) The pharmaceutical composition according to (1-6) or (1-7), which is in an eye drop dosage form.
(2-1) A Tie2 activator for use in reducing intraocular pressure.
(2-2) The Tie2 activator according to (2-1), which activates Tie2 in Schlemm's canal.
(2-3) The Tie2 activator according to (2-1) or (2-2), which is an extract of Siberian carrot and / or Keihi.
(2-4) The Tie2 activator according to any one of (2-1) to (2-3), which is an alcohol extract of Siberian carrot and / or Keihi.
(2-5) The Tie2 activator according to any one of (2-1) to (2-4), for preventing and / or treating glaucoma.
(2-6) The Tie2 activator according to any one of (2-1) to (2-5), wherein the Tie2 activator is administered orally.
(2-7) The Tie2 activator according to any one of (2-1) to (2-5), wherein the Tie2 activator is administered as an eye drop.
(3-1) A method for reducing intraocular pressure by administering a Tie2 activator to a subject who requires a reduction in intraocular pressure.
(3-2) The method according to (3-1), wherein the Tie2 activator activates Tie2 in Schlemm's canal.
(3-3) The method according to (3-1) or (3-2), wherein the Tie2 activator is an extract of Siberian ginseng and / or Keihi.
(3-4) The method according to any one of (3-1) to (3-3), wherein the Tie2 activator is an alcohol extract of Siberian ginseng and / or Keihi.
(3-5) The method according to any one of (3-1) to (3-4), for preventing and / or treating glaucoma.
(3-6) The method according to any one of (3-1) to (3-5), wherein the Tie2 activator is administered orally.
(3-7) The method according to any one of (3-1) to (3-5), wherein the Tie2 activator is administered as an eye drop.
　本発明によれば、Tie2活性化剤の投与により、シュレム管のTie2を活性化し、眼圧を低下させることができる。本発明によれば、Tie2活性化剤からなる眼圧降下剤を含有する医薬組成物やサプリメント等の食品組成物を、緑内障を予防及び／又は治療するために用いることができる。 According to the present invention, administration of the Tie2 activator can activate Tie2 in Schlemm's canal and reduce intraocular pressure. According to the present invention, a food composition such as a pharmaceutical composition or a supplement containing an intraocular pressure-lowering agent comprising a Tie2 activator can be used for preventing and / or treating glaucoma.
図１は、シュレム管におけるTie2の発現状態を示す。図１Ａは、眼の解剖学的模式図を示す。図１Ｂは、ＨＥ染色した眼球組織切片を示す(4倍)。図１Ｃは、図１Ｂを拡大したものを示し(10倍)、矢印はシュレム管を指す。図１Ｄは、同じ組織をanti-Tie2抗体で染色したものを示し、矢印はanti-Tie2抗体で染色されたシュレム管を指す。図１Ｅは、同じ組織をHoechstで核染色したものを示し、矢印はシュレム管を指す。FIG. 1 shows the expression state of Tie2 in Schlemm's canal. FIG. 1A shows an anatomical schematic diagram of the eye. FIG. 1B shows a HE-stained ocular tissue section (4 ×). FIG. 1C shows an enlargement of FIG. 1B (10 times), and the arrow points to Schlemm's canal. FIG. 1D shows the same tissue stained with anti-Tie2 antibody, with arrows pointing to Schlemm's canal stained with anti-Tie2 antibody. FIG. 1E shows nuclear staining of the same tissue with Hoechst, with arrows pointing to Schlemm's canal. 図２は、シベリアニンジン抽出物を朝９時に経口投与した場合の、９時と１７時における眼圧を示す。FIG. 2 shows intraocular pressure at 9:00 and 17:00 when the Siberian carrot extract was orally administered at 9:00 am. 図３は、シベリアニンジン抽出物を午前８時に経口投与した場合の、各時間における眼圧を示す。FIG. 3 shows intraocular pressure at each time when the Siberian carrot extract was orally administered at 8 am. 図４は、ケイヒ抽出物を朝９時に経口投与した場合の、９時と１７時における眼圧を示す。FIG. 4 shows intraocular pressure at 9:00 and 17:00 when the cinnamon extract is orally administered at 9:00 am.
　一つの態様では、本発明は、Tie2活性化剤からなる眼圧降下剤に関する。また、本発明は、Tie2活性化剤からなる眼圧降下剤を含有する医薬組成物やサプリメント等の食品組成物に関する。本発明の医薬組成物やサプリメント等の食品組成物は、極めて優れた眼圧降下作用を有するので、緑内障、高眼圧症の予防及び／又は治療するために用いることができる。これまでに知られている眼圧降下剤としては、交感神経刺激薬、交感神経遮断薬、副交感神経刺激薬、プロスタグランジン関連薬、炭酸脱水素酵素阻害薬などがあるが、本発明のようにシュレム管におけるTie2を活性化することにより、房水の適切な排出を促す眼圧降下剤はこれまで知られていない。 In one aspect, the present invention relates to an intraocular pressure-lowering agent comprising a Tie2 activator. The present invention also relates to food compositions such as pharmaceutical compositions and supplements containing an intraocular pressure-lowering agent comprising a Tie2 activator. Since the food composition such as the pharmaceutical composition or supplement of the present invention has an extremely excellent intraocular pressure-lowering action, it can be used for the prevention and / or treatment of glaucoma and ocular hypertension. Conventionally known intraocular pressure-lowering agents include sympathomimetic drugs, sympathetic nerve blockers, parasympathomimetic drugs, prostaglandin-related drugs, carbonic acid dehydrogenase inhibitors, and the like. In the past, no intraocular pressure-lowering agent that promotes proper drainage of aqueous humor by activating Tie2 in Schlemm's canal is known.
　Tie2の活性化剤は特に限定されるものではないが、アンジオポエチン1など、Tie2を活性化する活性を有することが周知のものや、ニッケイ（Cinnamomum)属植物由来の抽出物、シベリアニンジン（Siberian Ginseng）抽出物あるいはシリンガレシノール、エレウテロシドＥ（Eleutheroside）、エレウテロシドＥ１といった、本発明者がその活性を有することを見出したものが挙げられる。なお、ここでいうTie2の活性化とはTie2をリン酸化することでその活性体（リン酸化Tie2）に変換できる能力をいう。 The activator of Tie2 is not particularly limited, but is known to have an activity to activate Tie2, such as Angiopoietin 1, or an extract derived from a plant of the genus Cinnamomum, Siberian carrot (SiberianiberGinseng) ) Extracts or syringalesinol, eleutheroside E (Eleutheroside), eleutheroside E1, and the like that the present inventors have found to have the activity. The activation of Tie2 here refers to the ability to phosphorylate Tie2 and convert it into its active form (phosphorylated Tie2).
　ニッケイ属はクスノキ目（Lauraceae)、クスノキ科（Lauraceae)の植物であり、300以上の種が存在し、例えばケイ（Cinnamomum cassia Blume)、クスノキ（C. camphora）、マルバニッケイ（C. daphnoides）シバニッケイ（C. doederleinii）、ヤブニッケイ（C. japonicum）、オガサワラヤブニッケイ（C. pseudo-pedunculatum）、ニッケイ（C. sieboldii）、シバヤブニッケイ(Cinnamomum x takushii)、セイロンニッケイ（C. verum）、シナモン（C. zeylanicum）が知られる。本発明におけるTie2活性化剤として好ましくはケイ（Cinnamomum cassia Blume)、特にケイの若枝であるケイシ（桂枝）又はケイヒ（桂皮）由来の抽出物が使用される。 The genus Nikkei is a plant of the order of Lauraceae and Lauraceae, and there are more than 300 species, such as Cinnamomum cassia Blume, C. camphora, C. daphnoides (C. doederleinii), C. japonicum, Ogasawara jay nick (C. pseudo-pedunculatum), Nikkei (C. zeylanicum) is known. As the Tie2 activator in the present invention, an extract derived from Cay (Cinnamomum cassia Blume), in particular, Cay nae (Keishi) or Keihi (Cinnamon), which is a young branch of Cay, is used.
　シベリアニンジン（Siberian Ginseng）は、エゾウコギとも呼ばれ、ウコギ科に属する植物である。薬用部分は根皮であり、古くから漢方や生薬の原料として使われてきている。 Siberian carrot (Siberian Ginseng) is also called Ezoukogi and is a plant belonging to the family Argiaceae. The medicinal part is the root bark, which has long been used as a raw material for traditional Chinese medicine and herbal medicines.
　上記抽出物は常法により得ることができ、例えばその起源となる植物を抽出溶媒とともに常温又は加熱して浸漬または加熱還流した後、濾過し、濃縮して得ることができる。抽出溶媒としては、通常抽出に用いられる溶媒であれば任意に用いることができ、例えば、水性溶媒、例えば水、生理食塩水、リン酸緩衝液、ホウ酸緩衝液、あるいは有機溶媒、例えばエタノール、プロピレングリコール、1,3-ブチレングリコール、グリセリン等のアルコール類、含水アルコール類、クロロホルム、ジクロルエタン、四塩化炭素、アセトン、酢酸エチル、ヘキサン等を、それぞれ単独あるいは組み合わせて用いることができる。好ましくは、溶媒として水が使用される。上記溶媒で抽出して得られた抽出物をそのまま、あるいは例えば凍結乾燥などにより濃縮したエキスを使用でき、また必要であれば吸着法、例えばイオン交換樹脂を用いて不純物を除去したものや、ポーラスポリマー（例えばアンバーライトXAD-2）のカラムにて吸着させた後、所望の溶媒で溶出し、さらに濃縮したものや活性成分のみも使用することができる。 The above-mentioned extract can be obtained by a conventional method, for example, it can be obtained by soaking or heating and refluxing the plant that is the origin together with the extraction solvent at room temperature or heating, followed by filtration and concentration. The extraction solvent can be arbitrarily used as long as it is a solvent that is usually used for extraction. For example, an aqueous solvent such as water, physiological saline, phosphate buffer, borate buffer, or an organic solvent such as ethanol, Alcohols such as propylene glycol, 1,3-butylene glycol and glycerin, hydrous alcohols, chloroform, dichloroethane, carbon tetrachloride, acetone, ethyl acetate, hexane and the like can be used alone or in combination. Preferably, water is used as the solvent. The extract obtained by extraction with the above solvent can be used as it is or, for example, an extract concentrated by lyophilization or the like, and if necessary, an adsorbent method, for example, an ion exchange resin removed impurities, After adsorption on a polymer column (for example, Amberlite XAD-2), elution with a desired solvent, and further concentrated or only active ingredients can be used.
　本発明の医薬組成物の投与形態は、経口、非経口、外用等であってよい。剤型としては、例えば錠剤、粉剤、カプセル剤、顆粒剤、エキス剤、シロップ剤等の経口投与剤、又は注射剤、点滴剤、点眼剤、若しくは坐剤等の非経口投与剤軟膏、クリーム、乳液、ローション等の外用剤を挙げることができる。食品組成物の投与形態としてはサプリメント、顆粒剤、ドリンク剤等を挙げることができる。 The dosage form of the pharmaceutical composition of the present invention may be oral, parenteral, external use and the like. Examples of the dosage form include oral administration agents such as tablets, powders, capsules, granules, extracts and syrups, or parenteral ointments such as injections, drops, eye drops, or suppositories, creams, Mention may be made of external preparations such as emulsions and lotions. Examples of the dosage form of the food composition include supplements, granules, drinks and the like.
　本発明の医薬組成物中や食品組成物中のTie2活性化剤の配合量は、用途に応じて適宜決定できるが、一般には剤全量中、0.0001～20.0モル％、好ましくは0.0001～10.0モル％である。 The blending amount of the Tie2 activator in the pharmaceutical composition or food composition of the present invention can be appropriately determined depending on the use, but is generally 0.0001 to 20.0 mol%, preferably 0.0001 to 10.0 mol% in the total amount of the agent. It is.
　また、本発明の医薬組成物中や食品組成物中には、例えば、通常の食品や医薬品に使用される賦形剤、防湿剤、防腐剤、強化剤、増粘剤、乳化剤、酸化防止剤、甘味料、酸味料、調味料、着色料、香料等を必要に応じて適宜配合することができる。 In the pharmaceutical composition and food composition of the present invention, for example, excipients, moisture-proofing agents, preservatives, reinforcing agents, thickeners, emulsifiers, antioxidants used in ordinary foods and pharmaceuticals. , Sweeteners, acidulants, seasonings, colorants, fragrances and the like can be appropriately blended as necessary.
　本発明の眼用医薬組成物は、その剤形に依存して慣用の医薬的に許容される担体を併用することができる。 The ophthalmic pharmaceutical composition of the present invention can be used in combination with a conventional pharmaceutically acceptable carrier depending on the dosage form.
　本発明の眼用医薬組成物には、任意的に、糖質、電解質、アミノ酸、ビタミン、脂質、医薬品添加物、医薬品等の各種成分が配合されていてよい。そのような成分の例には、例えば、糖類、例えばグルコース、マルトース等、オリゴ糖、マンニトール、糖アルコール類、例えばソルビトール等；電解質、例えば塩化ナトリウム、リン酸水素ナトリウム、塩化カリウム、硫酸マグネシウム、塩化カルシウム等；アミノ酸、例えばグリシン、アラニン等； ビタミン類、例えば塩酸チアミン、リン酸リボフラビンナトリウム、塩酸ピリドキシン、ニコチン酸アミド、葉酸、ビオチン、ビタミンＡ 、Ｌ－ アスコルビン酸、α－グリコシルアスコルビン酸等及びそれらの誘導体が挙げられ、必要に応じてこれらを適宜組合せて配合してよい。 The ophthalmic pharmaceutical composition of the present invention may optionally contain various components such as carbohydrates, electrolytes, amino acids, vitamins, lipids, pharmaceutical additives, and pharmaceuticals. Examples of such components include, for example, sugars such as glucose, maltose, oligosaccharides, mannitol, sugar alcohols such as sorbitol; electrolytes such as sodium chloride, sodium hydrogen phosphate, potassium chloride, magnesium sulfate, chloride Calcium, etc .; amino acids such as glycine, alanine, etc .; vitamins such as thiamine hydrochloride, sodium riboflavin phosphate, pyridoxine hydrochloride, nicotinamide, folic acid, biotin, vitamin A, L-ascorbic acid, α-glycosyl ascorbic acid and the like These derivatives may be mentioned, and these may be combined in combination as necessary.
　本発明の眼用医薬組成物としては、通常使用される添加剤、例えば保存剤、例えばパラオキシ安息香酸メチル、デヒドロ酢酸ナトリウム、塩化ベンザルコニウム等；安定化剤、例えばエデト酸ナトリウム、亜硫酸水素ナトリウム等；緩衝剤、例えば硼砂、硼酸、炭酸水素ナトリウム等；増粘剤、例えばメチルセルロース、カルボキシメチルセルロース、コンドロイチン硫酸、ポリビニルアルコール、プルラン等；溶解補助剤、例えばポリソルベート80等；を配合してよい。 Examples of the ophthalmic pharmaceutical composition of the present invention include commonly used additives such as preservatives such as methyl parahydroxybenzoate, sodium dehydroacetate, benzalkonium chloride and the like; stabilizers such as sodium edetate and sodium bisulfite. Buffers such as borax, boric acid, sodium hydrogen carbonate, etc .; thickeners such as methyl cellulose, carboxymethyl cellulose, chondroitin sulfate, polyvinyl alcohol, pullulan, etc .; solubilizers such as polysorbate 80 may be blended.
　本発明の眼用医薬組成物が緑内障治療又は予防薬として用いるときは、コリン系眼圧降下剤(例えばピロカルピン、縮瞳効果に優れたカルバコール等)、抗コリンステラーゼ(例えばデメカリウム、D.F.P.、エコチオフェート等)、縮瞳剤としてのサリチル酸フイゾスチグミン、塩酸ピロカルピン等、静注用高浸透圧剤としてのマンニトール、グリセリン、イソソルバイド等、点眼剤用防腐剤としてのクロロブタノール、ベンズアルコニウムクロリド、プロピルパラベン、メチルパラベン、エチルパラベン、ブチルパラベン、他の炎症疾患予防および治療に用いられるペニシリン、サルファ剤、クロラムフェニコール、コルチネゾン、クロルフェニラミン等を配合してもよい。 When the ophthalmic pharmaceutical composition of the present invention is used as a glaucoma therapeutic or preventive agent, a cholinergic intraocular pressure-lowering agent (e.g., pilocarpine, carbachol excellent in miosis effect), anticholinesterase (e.g., deme potassium, DFP, ecothiol) Fate, etc.), physostigmine salicylate as a miotic agent, pilocarpine hydrochloride, mannitol, glycerin, isosorbide, etc. as intravenous hyperosmotic agents, chlorobutanol, benzalkonium chloride, propylparaben as antiseptics for eye drops, Methylparaben, ethylparaben, butylparaben, penicillin, sulfa drugs, chloramphenicol, cortinonezone, chlorpheniramine and the like used for the prevention and treatment of other inflammatory diseases may be added.
　次に実施例によって本発明をさらに詳細に説明する。なお、本発明はこれにより限定されるものではない。 Next, the present invention will be described in more detail with reference to examples. In addition, this invention is not limited by this.
実験１：シュレム管におけるTie2の発現
　マウス眼球をパラホルムアルデヒド固定した後、6μmの凍結切片をスライドガラス上で作製した。一次抗体をanti-Tie2抗体（Santa Cruz Biotechnology、希釈倍率200倍）、2次抗体をAlexa Fluor594で標識されたanti-rabbit IgGにて発色した。蛍光顕微鏡（オリンパス）にて観察し、写真を取得した。
　図１ＣはＨＥ染色した眼球組織切片を拡大したものであり、矢印はシュレム管を指す。図１Ｄは同じ組織をanti-Tie2抗体で染色したものであり、矢印はanti-Tie2抗体で染色されたシュレム管を指す。シュレム管が染色されたことにより、シュレム管においてTie2が発現していることが分かる。 Experiment 1: Expression of Tie2 in Schlemm's canal After mouse eyes were fixed with paraformaldehyde, frozen sections of 6 µm were prepared on a glass slide. The primary antibody was developed with anti-Tie2 antibody (Santa Cruz Biotechnology, dilution factor 200 times), and the secondary antibody was developed with anti-rabbit IgG labeled with Alexa Fluor594. Observation was made with a fluorescence microscope (Olympus), and a photograph was obtained.
FIG. 1C is an enlarged view of a HE-stained eyeball tissue section, and the arrow points to Schlemm's canal. FIG. 1D shows the same tissue stained with anti-Tie2 antibody, and the arrow indicates Schlemm's canal stained with anti-Tie2 antibody. The staining of Schlemm's canal indicates that Tie2 is expressed in Schlemm's canal.
実験２：眼圧測定試験１（シベリアニンジン）
　対象者はゴールドマン圧平眼圧計にて両眼の眼圧が22mmHg以下、かつ細隙灯顕微鏡検査および眼底検査にて明らかな異常を認めず、眼圧・眼球血流に影響を及ぼす眼疾患・手術歴の既往や高血圧症や糖尿病などの全身疾患の既往、喫煙歴、点眼薬・内服歴のない成人の健常者とする。Tie2活性化剤として、シベリアニンジンの根を30％エタノールで抽出したシベリアニンジン抽出物（アスク薬品）であるシベリア人参粉末を使用した。
　サンプルサイズを20名（男女各10名）と予測し、全対象者についてシベリア人参粉末を投与する。洗い流し期間を1ヶ月以上あけ、3回の内服と前後における眼圧や血圧、脈拍、レーザースペックル検査を行う。主要エンドポイントは眼圧の低下であり、副次エンドポイントはレーザースペックルフローグラフィにより計測された眼球血流の増加である。これらエンドポイントに対しシベリア人参粉末について統計解析（ANOVA）をおこなった。
　図２は、摂取時（午前9時）と17時における眼圧を示す。シベリア人参粉末摂取後、3時間程度で眼圧の低下が見られ、8時間程度持続した。なお、コントロールはシベリア人参粉末を摂取していない同一の対象者とし、眼圧の測定はシベリア人参粉末の摂取日とは異なる日に行った。 Experiment 2: Intraocular pressure measurement test 1 (Siberian carrot)
The subject had an intraocular pressure of 22 mmHg or less with a Goldman applanation tonometer, and no ocular abnormalities were observed in slit lamp microscopy and fundus examination.・ Health history, history of systemic diseases such as hypertension and diabetes, smoking history, and healthy adults with no eye drops or internal history. As a Tie2 activator, Siberian ginseng powder, a Siberian carrot extract obtained by extracting 30% ethanol from Siberian carrot roots (Ask Chemical), was used.
Expect a sample size of 20 (10 men and women) and administer Siberian ginseng powder to all subjects. The wash-out period is longer than 1 month, and three internal doses and before and after intraocular pressure, blood pressure, pulse, and laser speckle tests are performed. The primary endpoint is a decrease in intraocular pressure, and the secondary endpoint is an increase in ocular blood flow as measured by laser speckle flowography. Statistical analysis (ANOVA) was performed on Siberian ginseng powder at these endpoints.
FIG. 2 shows intraocular pressure at the time of ingestion (9 am) and 17:00. After ingestion of Siberian ginseng powder, a decrease in intraocular pressure was observed in about 3 hours, which lasted for about 8 hours. The control was the same subject who did not take Siberian ginseng powder, and the intraocular pressure was measured on a different day from the day of taking Siberian ginseng powder.
実験３：眼圧測定試験２（シベリアニンジン）
　内服時を午前8時とし、非接触型眼圧計により眼圧を測定した以外は、実験２と同様の条件を用いた。
　図３は、各時間における眼圧を示す。シベリア人参粉末摂取後、10時間程度まで眼圧の低下が見られた。 Experiment 3: Intraocular pressure measurement test 2 (Siberian carrot)
The same conditions as in Experiment 2 were used except that the internal use was 8:00 am and the intraocular pressure was measured with a non-contact tonometer.
FIG. 3 shows intraocular pressure at each time. After taking Siberian ginseng powder, the intraocular pressure decreased until about 10 hours.
実験４：眼圧測定試験（ケイヒ）
　対象者はゴールドマン圧平眼圧計にて両眼の眼圧が22mmHg以下、かつ細隙灯顕微鏡検査および眼底検査にて明らかな異常を認めず、眼圧・眼球血流に影響を及ぼす眼疾患・手術歴の既往や高血圧症や糖尿病などの全身疾患の既往、喫煙歴、点眼薬・内服歴のない成人の健常者とする。Tie2活性化剤として、ケイヒの樹皮を３０％エタノールで抽出したケイヒエキスパウダー（日本粉末薬品株式会社）を使用した。
　サンプルサイズを８名（女性5名・男性3名）とし、全対象者について午前９時を投与時刻としてケイヒエキスパウダーを１００ｍｇ摂取させ（市販のミネラルウォーターとともに：商品名；い・ろ・は・す）（登録商標）、ゴールドマン圧平眼圧計により眼圧を測定した。洗い流し期間を1ヶ月以上あけ、１回の内服と前後における眼圧や血圧、脈拍、レーザースペックル検査を行った。主要エンドポイントは眼圧の低下であり、副次エンドポイントはレーザースペックルフローグラフィにより計測された眼球血流の増加である。これらエンドポイントに対しケイヒ粉末について統計解析（ANOVA）をおこなった。
　図４は、各時間における左眼の眼圧を示す。ケイヒ粉末摂取後、3時間で有意に眼圧の減少が見られた。このような眼圧の低下は、ケイヒ粉末摂取後１０時間程度まで見られた。なお、コントロールはケイヒ粉末を摂取していない同一の対象者とし、眼圧の測定はケイヒ粉末の摂取日とは異なる日に行った。 Experiment 4: Intraocular pressure measurement test (Keihi)
The subject had an intraocular pressure of 22 mmHg or less with a Goldman applanation tonometer, and no ocular abnormalities were observed in slit lamp microscopy and fundus examination.・ Health history, history of systemic diseases such as hypertension and diabetes, smoking history, and healthy adults with no eye drops or internal history. As a Tie2 activator, Keihi extract powder (Nippon Powder Chemical Co., Ltd.) obtained by extracting Keihi bark with 30% ethanol was used.
The sample size is 8 (5 women and 3 men), and 100 mg of keihi extract powder is ingested at 9 am for all subjects (with commercial mineral water: product name; The intraocular pressure was measured with a Goldman applanation tonometer. The wash-out period was 1 month or longer, and a single oral dose and before and after intraocular pressure, blood pressure, pulse, and laser speckle tests were performed. The primary endpoint is a decrease in intraocular pressure, and the secondary endpoint is an increase in ocular blood flow as measured by laser speckle flowography. Statistical analysis (ANOVA) was conducted on Keihi powder for these endpoints.
FIG. 4 shows the intraocular pressure of the left eye at each time. There was a significant decrease in intraocular pressure 3 hours after ingestion of Keihi powder. Such a decrease in intraocular pressure was observed up to about 10 hours after ingestion of Keihi powder. The control was the same subject who did not take keihi powder, and the intraocular pressure was measured on a different day from the day of taking keihi powder.

Claims (9)

  1. 　Tie2（tyrosine kinase with Ig and EGF homology domain2）活性化剤からなる、眼圧降下剤。 An intraocular pressure-lowering agent consisting of Tie2 (tyrosine kinase with Ig and EGF homology domain 2) activator
  2. 　前記Tie2活性化剤が、シュレム管におけるTie2を活性化する、請求項１に記載の眼圧降下剤。 The intraocular pressure-lowering agent according to claim 1, wherein the Tie2 activator activates Tie2 in Schlemm's canal.
  3. 　前記Tie2活性化剤が、シベリアニンジン（Siberian Ginseng）及び／又はケイヒの抽出物である、請求項１又は２に記載の眼圧降下剤。 The intraocular pressure-lowering agent according to claim 1 or 2, wherein the Tie2 activator is an extract of Siberian ginseng and / or Keihi.
  4. 　前記Tie2活性化剤が、シベリアニンジン（Siberian Ginseng）及び／又はケイヒのアルコール抽出物である、請求項１～３のいずれか一項に記載の眼圧降下剤。 The intraocular pressure-lowering agent according to any one of claims 1 to 3, wherein the Tie2 activator is an alcohol extract of Siberian ginseng and / or Keihi.
  5. 　請求項１～４のいずれか一項に記載の眼圧降下剤を含有する、組成物。 A composition comprising the intraocular pressure-lowering agent according to any one of claims 1 to 4.
  6. 　請求項１～４のいずれか一項に記載の眼圧降下剤を含有する、医薬組成物。 A pharmaceutical composition comprising the intraocular pressure-lowering agent according to any one of claims 1 to 4.
  7. 　緑内障を予防及び／又は治療するための、請求項６に記載の医薬組成物。 The pharmaceutical composition according to claim 6, for preventing and / or treating glaucoma.
  8. 　経口投与の剤型にある、請求項６又は７に記載の医薬組成物。 The pharmaceutical composition according to claim 6 or 7, which is in a dosage form for oral administration.
  9. 　点眼剤の剤型にある、請求項６又は７に記載の医薬組成物。 The pharmaceutical composition according to claim 6 or 7, which is in an eye drop dosage form.
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