HIF inhibitory composition

Abstract

To provide a composition for inhibiting HIFs and effectively improving related diseases and abnormalities in the eye.SOLUTION: Provided is an HIF inhibitory composition containing pyridoxine and a salt thereof. Also provided are a retina protection composition, an anti-aging composition for the eye, a VEGF inhibitory composition for the eye, a neovascularization inhibitory composition for the eye, a composition for maintaining the homeostasis of vascular tissues in the eye, and a composition for reducing the risk of, preventing and treating a disease in the posterior segment of the eye, each of which contains the above component.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a composition for suppressing HIF. The present invention also relates to a composition for protecting the retina, a composition for anti-aging of the eye, a composition for suppressing VEGF in the eye, a composition for suppressing angiogenesis in the eye, a composition for maintaining homeostasis of vascular tissue in the eye, and a composition for reducing the risk of, preventing, or treating posterior eye diseases. [Background technology]

[0002] HIF (Hypoxia Inducible Factor) is a transcription factor that is induced when cells are deprived of oxygen. It has a variety of physiological activities, including hypoxia adaptation response, maintenance of stem cells, and homeostasis such as control of inflammation. However, overexpression of HIF is thought to be closely related to the onset and progression of cancer.

[0003] For this reason, HIF is being studied as a target for cancer treatment, for example, as disclosed in Non-Patent Document 1. On the other hand, as disclosed in Patent Document 1 and Non-Patent Document 2, suppression of HIF is expected to have therapeutic, preventive, and risk-reducing effects not only on cancer, but also on ophthalmological diseases and ocular abnormalities. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2018 / 088566 [Non-patent literature]

[0005] [Non-Patent Document 1] Onnis B, et al. “Development of HIF-1 inhibitors for cancer therapy”JCell Mol Med. 2009 [Non-Patent Document 2] Kurihara T, et al. “Hypoxia-induced metabolic stress inretinal pigment epithelial cells is sufficient to induce photoreceptordegeneration” eLife2016 [Non-Patent Document 3] Wang X, et al. “Mitochondrialferritin affects mitochondria bystabilizing HIF-1arufwa in retinalpigmentepithelium: implicationsfworthe pathophysiologyof age-relatedmacular degeneration”Neurobiol Aging. 2016 Summary of the Invention [Problem to be solved by the invention]

[0006] However, as disclosed in Non-Patent Document 1, most of the compositions for suppressing HIF are anticancer drugs, and therefore have strong cytotoxicity, making it difficult to directly apply them to, for example, eye diseases. Therefore, there is a demand for novel compositions for suppressing HIF that use ingredients that are highly safe and can be applied not only in the field of pharmaceuticals but also in the field of functional foods and supplements.

[0007] Therefore, an object of the present invention is to provide a composition that suppresses HIF and thereby effectively improves associated diseases and ocular abnormalities. [Means for solving the problem]

[0008] In order to solve the above problems, the present inventors conducted intensive research and discovered that ingredients that are safely used in the food and pharmaceutical fields have been found to have HIF inhibitory activity, leading to the completion of the present invention.

[0009] That is, the present invention provides the compositions described below. [1] A composition for suppressing HIF, comprising at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine or its derivatives and their salts, niacin, inositol, phytic acid, pantothenic acid, biotin, and folic acid. [2] A retinal protective composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine or its derivatives and their salts, niacin, inositol, phytic acid, pantothenic acid, biotin, and folic acid. [3] An anti-aging composition for the eye, comprising at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine or its derivatives and their salts, niacin, inositol, phytic acid, pantothenic acid, biotin, and folic acid. [4] A composition for suppressing VEGF in the eye, comprising at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine or its derivatives and their salts, niacin, inositol, phytic acid, pantothenic acid, biotin, and folic acid. [5] A composition for inhibiting ocular angiogenesis, comprising at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine or its derivatives and their salts, niacin, inositol, phytic acid, pantothenic acid, biotin, and folic acid. [6] A composition for maintaining homeostasis of ocular vascular tissue, comprising at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine or its derivatives and their salts, niacin, inositol, phytic acid, pantothenic acid, biotin, and folic acid. [7] A composition for reducing the risk of, preventing, or treating a posterior eye disease, comprising at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine or its derivatives and their salts, niacin, inositol, phytic acid, pantothenic acid, biotin, and folic acid. [8] The composition according to [7], wherein the posterior ocular disease is at least one selected from the group consisting of age-related macular degeneration, diabetic retinopathy, diabetic macular edema, retinitis pigmentosa, proliferative vitreous retinopathy, retinal artery occlusion, retinal vein occlusion, uveitis, Leber's disease, retinopathy of prematurity, retinal detachment, retinal pigment epithelial detachment, central serous chorioretinopathy, central exudative chorioretinopathy, polypoidal choroidal vasculopathy, multifocal choroiditis, neovascular maculopathy, retinal aneurysm, retinal angiomatous proliferation, optic neuropathy caused by these diseases, optic neuropathy caused by glaucoma, and ischemic optic neuropathy. Effect of the Invention

[0010] According to the present invention, it is possible to inhibit HIF using ingredients that are safely used in the food and pharmaceutical fields, and to effectively improve ocular abnormalities and diseases caused by high levels of HIF. [Brief description of the drawings]

[0011] [Figure 1] 1(A) to (D) are graphs showing the results of ERG (rod system function) measurement by stimulation (10 cd s / m 2 ) under dark adaptation in Test Example 3. [Diagram 2]2(A) and (B) are graphs showing the results of ERG (cone system function) measurement by stimulation (10 cd s / m 2 ) under light adaptation (Lightadapted) in Test Example 3. [Diagram 3] 3(A) and (B) are graphs showing the results of ERG (rod system function) measurement by stimulation (10 cd s / m 2 ) under dark-adapted conditions in Test Example 3. [Figure 4] FIG. 4 is a graph showing the results of ERG (cone system function) measurement using light-adapted stimulation (10 cd s / m2) in Test Example 3. [Diagram 5] Fig. 5(A) is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation on the 42nd day (6 weeks) after the start of administration in Test Example 4. Fig. 5(B) is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation on the 56th day (8 weeks) after the start of administration in Test Example 4. [Figure 6] FIG. 6 is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation in Test Example 5. [Figure 7] FIG. 7 is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation in Test Example 6. [Figure 8] FIG. 8 is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation in Test Example 7. [Figure 9] FIG. 9 is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation in Test Example 8. [Figure 10] FIG. 10 is a graph showing the results of confirming the expression of downstream genes of HIF in Test Example 9. [Figure 11] FIG. 11 is a graph showing the results of confirming the expression of downstream genes of HIF in Test Example 9. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] [HIF suppression composition] In one embodiment, the HIF-suppressing composition of the present invention contains at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, and oleanolic acid and its salts.

[0013] In the present specification, the plant extract may be a crude extract extracted from the whole plant or a necessary part of the plant (flower, inflorescence, flower bud, flower spike, leaf, branch, branch leaf, rhizome, root bark, root, bark, fruit, pericarp, legume, seed, etc.), or it may be a product obtained by further purification or concentration, or may be a product obtained by synthesis, or a commercially available product may be used. The method for obtaining the plant extract is not particularly limited, and a usual extraction method, purification method, concentration method, synthesis method, drying powder method, etc. may be used.

[0014] (Hydroxycitric acid and its salts) Hydroxycitric acid (HCA; 1,2-dihydroxypropane-1,2,3-tricarboxylic acid) is a natural substance found naturally in the pericarp of plants of the Garcinia genus, such as Garcinia cambogia. Its calcium salt (calcium hydroxycitrate) is known to inhibit fatty acid biosynthesis. Garcinia cambogia is a plant of the Hypericaceae family native to Southeast Asia that has long been used as a spice and preservative, and is now well known as a diet ingredient due to its ability to inhibit fat synthesis. Various products containing HCA, including pharmaceuticals, have been developed that claim to have anti-obesity effects.

[0015] The salt of hydroxycitric acid is not particularly limited as long as it is medicamentarily, pharmacologically (pharmaceutical) or physiologically acceptable, and examples thereof include alkali metal (particularly sodium) salts and alkaline earth metal (particularly calcium or magnesium) salts, etc. As the salt of hydroxycitric acid, calcium salts are preferred.

[0016] These hydroxycitric acid and / or its salts may be used alone or in any combination of two or more. In addition, food materials containing hydroxycitric acid and / or its salts may be used. Such materials include, but are not limited to, Garcinia cambogia and its extract.

[0017] (Garcinia and its extract) Garcinia and its extract refers to the fruit or peel of Garcinia cambogia, an evergreen tree of the Hypericaceae family, its dried matter, its fine powder, and its extract.

[0018] The Garcinia extract is not limited by the extraction method as long as it can achieve the effects of the present invention. For example, the Garcinia extract refers to an extract obtained by immersing Garcinia or its crushed product in water and / or an organic solvent and filtering the residue, a product obtained by removing the solvent from the extract, or a fine powder of these, or a product obtained by dissolving, dispersing, or diluting the above extract or the product obtained by removing the solvent using an appropriate solvent, and a commercially available product can also be used. As another example of the extraction method, the Garcinia extract can be extracted by processing the fruit or peel of Garcinia cambogia by steaming or the like.

[0019] In the present specification, when using a plant extract (also referred to as a plant extract), more specifically, as the extraction solvent, water (including hot water), alcohols such as methanol, ethanol, isopropanol, ethylene glycol, 1,3-butylene glycol, glycerin, etc., esters such as ethyl acetate, ketones such as acetone and methyl ethyl ketone, nitriles such as acetonitrile, ethers such as diethyl ether and tetrahydrofuran, saturated hydrocarbons such as pentane, hexane, cyclopentane, cyclohexane, aromatic hydrocarbons such as toluene, halogenated hydrocarbons such as dichloromethane and chloroform, and other organic solvents such as dimethylformamide and dimethyl sulfoxide (which may all contain water) can be appropriately used, and may be any mixture of one or two of them. Among these solvents, water, ethanol, 1,3-butylene glycol, or a mixture of these is preferable. The extracts described in the present specification can be obtained from various raw material companies, and are usually sold in a form including excipients, but are not limited thereto. Hereinafter, the amount of extract refers to the amount including the dry solid content and excipients.

[0020] The extraction solvent for Garcinia extract is not limited as long as it achieves the effects of the present invention, but is preferably water, ethanol, or aqueous ethanol, and more preferably aqueous ethanol.

[0021] Examples of food materials containing hydroxycitric acid or commercially available Garcinia extract products include, but are not limited to, Super Citrimax (Ryusendo Co., Ltd.), Garcinia Cambogia Extract Powder (Vidya Japan Co., Ltd.), Garcinia Cambogia (BioActives Japan Co., Ltd.), Garcitrin (Sabinsa Japan Corporation), and Garcinia Extract Powder (Koshin Bussan Co., Ltd.).

[0022] (Lactoferrin) Lactoferrin is a glycoprotein with a molecular weight of approximately 80,000 daltons, which can reversibly bind two iron atoms per molecule, and is present in mammalian milk.

[0023] The lactoferrin used in the present invention includes lactoferrin obtained by separating and purifying the raw material from the milk (colostrum, transitional milk, normal milk, terminal milk, etc.) of mammals (e.g., humans, cows, goats, sheep, horses, etc.) or processed products of these milks (skim milk, whey, etc.) by a conventional method (e.g., ion exchange chromatography, etc.) (isolated lactoferrin); apolactoferrin obtained by removing iron from the isolated lactoferrin with hydrochloric acid, citric acid, etc.; various lactoferrins produced by genetic engineering in microorganisms, animal cells, transgenic animals, etc. These lactoferrins may be used alone or as a mixture of two or more kinds. Commercially available lactoferrins may also be used.

[0024] Food materials containing lactoferrin may also be used. Examples of food materials containing lactoferrin include, but are not limited to, commercially available products containing lactoferrin, such as Lactoferrin (Rivason Co., Ltd.), Aura Barrier LF+LPO (Morinaga Milk Industry Co., Ltd.), IDP (Kanematsu Chemical Co., Ltd.), and Bioferrin (Glanbia Japan Co., Ltd.).

[0025] (Carrot and its extract) Carrots and their extracts refer to carrots, finely powdered carrots (carrot powder), and their extracts.

[0026] Specifically, ginseng powder is "ginseng" powder as defined in the Japanese Pharmacopoeia, and contains 0.10% or more of ginsenoside Rg1 and 0.20% or more of ginsenoside Rb1 based on the calculated dry matter of the herb. Ginseng powder that meets the standards set out in the Japanese Pharmacopoeia is commercially available and can be used.

[0027] The carrot extract is not limited by the extraction method as long as it can achieve the effects of the present invention. For example, the carrot extract refers to an extract obtained by immersing and extracting carrots or pulverized carrots in water and / or an organic solvent and filtering the residue, a product from which the solvent has been removed, or a fine powder of these, or a product obtained by dissolving, dispersing, or diluting the above-mentioned extract or the product from which the solvent has been removed using an appropriate solvent, and a commercially available product can also be used. As another example of the extraction method, the carrot extract can be extracted after processing carrots, such as steaming them.

[0028] The extraction solvent for the carrot extract is not limited as long as it achieves the effects of the present invention, but is preferably water, ethanol, or aqueous ethanol, and more preferably aqueous ethanol.

[0029] Furthermore, the origin of the ginseng and its extract is not limited as long as the effects of the present invention are achieved; however, it is preferable that the ginseng and its extract are from Panax ginseng (Korean ginseng), American ginseng (Western ginseng), Siberian ginseng (Eleuthero), Sanshichi ginseng (Tanshichi ginseng, Denshichi ginseng), or a combination of these, and it is more preferable that the ginseng is Panax ginseng (Korean ginseng).

[0030] Commercially available products of ginseng and its extracts include, but are not limited to, the following. Examples of Panax ginseng include black ginseng extract powder (Nihon Yakuhin Co., Ltd.), Korean ginseng extract powder (Frontier Foods Co., Ltd.), Korean ginseng extract (Sankt Co., Ltd.), ginseng extract powder S (Matsuura Yakugyo Co., Ltd.), and ginseng liquor extract 10% (Indena Japan Co., Ltd.). Examples of American ginseng include Western ginseng extract powder (FAP Japan Co., Ltd.), Western ginseng extract powder (Matsuura Yakugyo Co., Ltd.), Wisconsin ginseng (Kokan Pharmaceutical Research Institute Co., Ltd.), and American ginseng extract (Tama Biochemical Co., Ltd.). Examples of Siberian ginseng include Eleuthero extract (Matsuura Yakugyo Co., Ltd.), Siberian ginseng extract powder (Koshin Bussan Co., Ltd.), Eleuthero extract powder (FAP Japan Co., Ltd.), Eleuthero extract powder (Kokan Pharmaceutical Research Institute Co., Ltd.), and Siberian ginseng dried extract (Ask Pharmaceutical Co., Ltd.). Examples of Sanshichi ginseng (sanchi ginseng, danchi ginseng) include danchi ginseng extract powder (Nakahara Co., Ltd.), danchi ginseng extract powder (Matsuura Pharmaceutical Co., Ltd.), danchi ginseng (Adaptogen Pharmaceutical Co., Ltd.), etc., but commercially available products of ginseng and its extracts are not limited to these.

[0031] (Pyridoxine and its salts) Pyridoxine is a compound also known as 5-hydroxy-6-methylpyridine-3,4-dimethanol. Pyridoxine and / or its salts are water-soluble vitamins, including vitamin B 6 These compounds are known in the art and can be synthesized by known methods or are commercially available.

[0032] The salt of pyridoxine is not particularly limited so long as it is medicamentarily, pharmacologically (pharmaceutical) or physiologically acceptable, and specific examples thereof include organic acid salts [for example, monocarboxylates (acetate, trifluoroacetate, butyrate, palmitate, stearate, etc.), polycarboxylates (fumarate, maleate, succinate, malonate, etc.), oxycarboxylates (lactate, tartrate, citrate, etc.), organic sulfonates (methanesulfonate, toluenesulfonate, tosylate, etc.)], inorganic acid salts (for example, hydrochloride, sulfate, nitrate, hydrobromide, phosphate, etc.), and the like.

[0033] These pyridoxine and / or salts thereof may be used alone or in any combination of two or more. From the viewpoint of improving the foreign body sensation in the eye, the component (A) is preferably an inorganic acid salt of pyridoxine, more preferably a hydrochloride or phosphate of pyridoxine, and even more preferably a hydrochloride of pyridoxine.

[0034] Food materials containing pyridoxine and / or a salt thereof may also be used. Food materials containing pyridoxine and / or a salt thereof or commercially available products thereof include, for example, red pepper, rice bran, garlic, basil, parsley, wheat germ, etc., and commercially available products thereof include, for example, vitamin B 6 Contains yeast (Seti Co., Ltd.), vitamin B 6 (DSM Nutrition Japan Co., Ltd.), Capsicum Oleoresin (Vidya Japan Co., Ltd.), Selected Dried Defatted Rice Bran (Tsukuno Foods Industry Co., Ltd.), Oryza Dorim (Oryza Oil & Fat Chemical Co., Ltd.), Garlic Extract (BioActives Japan Co., Ltd.), etc., but are not limited to these.

[0035] Among food materials containing pyridoxine and / or a salt thereof, from the viewpoint of significantly exhibiting the effects of the present invention, rice bran is preferred, and defatted rice bran (also referred to as defatted rice bran extract) is more preferred.

[0036] Defatted rice bran is obtained by defatting rice bran obtained by polishing brown rice. It can also be used to make an extract with various extraction solvents. In the present invention, rice bran obtained from seeds of rice (Oryza sativa) belonging to the genus Oryza of the family Poaceae is used as a raw material, and rice bran derived from any variety of rice, such as japonica or indica, can be used.

[0037] (Oleanolic acid and its salts) Oleanolic acid is a type of β-amyrin triterpene, and is known to exist as a free form or saponin in olive leaves, Swertia japonica, Clove, Grape, Beet, Persimmon, Plum, Loquat, Sea Buckthorn, Argan, and Sugar Beet. Oleanolic acid can be extracted and purified from the above-mentioned raw materials by known methods, or it can be obtained by chemical synthesis or as a commercially available product.

[0038] Food materials containing oleanolic acid may also be used. Examples of food materials containing oleanolic acid or commercially available products thereof include, but are not limited to, Oleanolic-55 (Tama Biochemical Co., Ltd.), Oleavita (Nutrition Act Co., Ltd.), Nippum Pamis Extract GR, Nippum Pamis Extract OA (Nippon Flour Mills Co., Ltd.), and Palo Asul (IHM Co., Ltd.).

[0039] The oleanolic acid is not particularly limited as long as it is medicamentarily, pharmacologically (pharmaceutical) or physiologically acceptable, and examples thereof include alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt, and organic acid salts including basic amino acid salts such as arginine salt.

[0040] (Thiamin or its derivatives and their salts) Thiamine is a water-soluble vitamin B 1 These compounds are known in the art and can be synthesized by known methods or are commercially available.

[0041] The salt of thiamine is not particularly limited so long as it is medicamentarily, pharmacologically (pharmaceutical) or physiologically acceptable, and specific examples thereof include organic acid salts [for example, monocarboxylates (acetate, trifluoroacetate, butyrate, palmitate, stearate, etc.), polycarboxylates (fumarate, maleate, succinate, malonate, etc.), oxycarboxylates (lactate, tartrate, citrate, etc.), organic sulfonates (methanesulfonate, toluenesulfonate, tosylate, etc.)], inorganic acid salts (for example, hydrochloride, sulfate, nitrate, hydrobromide, phosphate, etc.), and the like.

[0042] These thiamine or its derivatives and their salts may be used alone or in any combination of two or more. Examples of thiamine or its derivatives and their salts include dibenzoylthiamine, dibenzoylthiamine hydrochloride, thiamine cetyl sulfate, thiamine thiocyanate, thiamine naphthalene-1,5-disulfonate, thiamine lauryl sulfate, bisbentiamine, etc. Among these, inorganic acid salts of thiamine are preferred, thiamine hydrochloride or nitrate are more preferred, and nitrate is even more preferred.

[0043] Food materials containing thiamine or its derivatives and salts thereof may also be used. Examples of food materials containing thiamine or its derivatives and salts thereof or commercially available products thereof include, but are not limited to, thiamine nitrate (Nakahara Co., Ltd.), thiamine hydrochloride (Sankt Co., Ltd.), vitamin B1-containing yeast (Seti Co., Ltd.), vitamin B1 (DSM Co., Ltd.), selected dried defatted rice bran (Tsukuno Foods Co., Ltd.), Oryza Dorim (Oryza Oil & Fat Chemical Co., Ltd.), germ powder (Nippon Flour Mills Co., Ltd.), and sesame (BioActives Japan Co., Ltd.).

[0044] (Niacin) Niacin is a general term for nicotinic acid (vitamin B3, niacin) and its amide compounds (nicotinamide, nicotinamide, niacinamide), which are known as water-soluble vitamins and can be synthesized by known methods or obtained commercially.

[0045] Niacin may be used alone or in any combination of two or more. From the viewpoint of improving the foreign body sensation in the eye, niacinamide is preferred as component (A).

[0046] Furthermore, food materials containing niacin may be used. Examples of food materials containing niacin or commercially available products thereof include, but are not limited to, rice bran, niacinamide-containing yeast (Cetie Co., Ltd.), nicotinic acid (DSM Co., Ltd.), selected dried defatted rice bran (Tsukuno Foods Co., Ltd.), and Oryza Dorim (Oryza Oil & Fat Chemical Co., Ltd.).

[0047] (Inositol) Inositol is a known compound that is a type of cyclitol and has a structure (1,2,3,4,5,6-cyclohexanehexaol) in which one hydrogen atom on each carbon of cyclohexane is replaced with a hydroxyl group. It can be synthesized by known methods or obtained commercially.

[0048] In addition, food materials containing inositol may be used. Examples of food materials containing inositol or commercially available products thereof include, but are not limited to, rice bran, inositol (Sankt Co., Ltd.), inositol (Tsukuno Foods Industry Co., Ltd.), selected dried defatted rice bran (Tsukuno Foods Industry Co., Ltd.), Oryzadrim (Oryza Oil & Fat Chemical Co., Ltd.), and Kittle Syrup (Pures Japan Co., Ltd.).

[0049] (Phytic acid and its salts) Phytic acid is a biological substance that is a hexaphosphate ester of myo-inositol, also called myo-inositol-1,2,3,4,5,6-hexaphosphate or hexakisphosphate or hexakis(dihydrogenphosphate), and can be synthesized by known methods or purchased commercially. Salts of phytic acid include calcium and magnesium salts of phytic acid (phytin), which are prepared from plant seeds and grains and are obtained as white powders that are almost insoluble in water.

[0050] Furthermore, food materials containing phytic acid may be used. Examples of food materials containing phytic acid or commercially available products thereof include, but are not limited to, rice bran, phytic acid (IP6) (Tsukuno Foods Industry Co., Ltd.), phytic acid FC (FC Chemical Co., Ltd.), phytic acid (Fuso Chemical Co., Ltd.), selected dried defatted rice bran (Tsukuno Foods Industry Co., Ltd.), Oryza Dorim (Oryza Oil & Fat Chemical Co., Ltd.), and the like.

[0051] (Pantothenic acid and / or its salts) Pantothenic acid, D(+)-N-(2,4-dihydroxy-3,3-dimethylbutyryl)-β-alanine, is a known compound that is part of the B vitamin group, which is a water-soluble vitamin. It can be synthesized by known methods or obtained commercially.

[0052] There are no particular limitations on the salt of pantothenic acid, so long as it is medicamentarily, pharmacologically (pharmaceutical) or physiologically acceptable, and specific examples include calcium salts, sodium salts, ammonium salts, etc.

[0053] These pantothenic acids and / or salts thereof may be used alone or in any combination of two or more. As component (A), calcium salts of pantothenic acid are preferred.

[0054] Food materials containing pantothenic acid and / or a salt thereof may also be used. Examples of food materials containing pantothenic acid and / or a salt thereof or commercially available products thereof include, but are not limited to, pantothenic acid (DSM Co., Ltd.), pantothenic acid-containing yeast (Seti Co., Ltd.), selected dried defatted rice bran (Tsukuno Foods Co., Ltd.), Oryza Dorim (Oryza Oil & Fat Chemical Co., Ltd.), and Capsicum Oleoresin (Vidya Japan Co., Ltd.).

[0055] (Biotin) It is a water-soluble vitamin classified as a B vitamin, and can be synthesized by known methods or obtained commercially.

[0056] Food materials containing biotin may also be used. Examples of food materials containing phytic acid or commercially available products thereof include, but are not limited to, rice bran, biotin-containing yeast (Cetie Co., Ltd.), selected dried defatted rice bran (Tsukuno Foods Co., Ltd.), and Oryza Dorim (Oryza Oil & Fat Chemical Co., Ltd.).

[0057] (Folic Acid) Folic acid is a water-soluble vitamin classified as a B vitamin, which has a structure in which pteridine is bound to para-aminobenzoic acid and one or more glutamic acids, and is a physiologically active substance classified as a water-soluble vitamin. It can be synthesized by known methods or is available commercially.

[0058] In addition, food materials containing folic acid may be used. Examples of food materials containing folic acid or commercially available products thereof include, but are not limited to, rice bran, folic acid-containing yeast (Cetie Co., Ltd.), and folic acid (DSM Co., Ltd.).

[0059] The total content of at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran is appropriately set depending on the types and contents of other blended ingredients and the formulation form of the composition, and is preferably 0.1 to 95% by mass, more preferably 0.3 to 80% by mass, even more preferably 0.5 to 75% by mass, and particularly preferably 1 to 70% by mass, based on the total amount of the composition.

[0060] When hydroxycitric acid and its salts are contained, the content of hydroxycitric acid and its salts alone is not particularly limited, but is, for example, preferably 0.1 to 95 mass%, more preferably 1 to 80 mass%, even more preferably 10 to 75 mass%, and particularly preferably 20 to 70 mass%, relative to the total amount of the composition.

[0061] When Garcinia and an extract thereof are contained, the content of Garcinia and an extract thereof alone is not particularly limited, but is, for example, preferably 0.1 to 95 mass% relative to the total amount of the composition, more preferably 1 to 80 mass%, even more preferably 10 to 75 mass%, and particularly preferably 20 to 70 mass%.

[0062] When carrot and its extract are contained, the content of carrot and its extract alone is not particularly limited, but is, for example, preferably 0.1 to 95 mass%, more preferably 1 to 80 mass%, even more preferably 10 to 75 mass%, and particularly preferably 20 to 70 mass%, relative to the total amount of the composition.

[0063] When pyridoxine and a salt thereof are contained, the content of pyridoxine and a salt thereof alone is not particularly limited, but is, for example, preferably 0.1 to 95 mass%, more preferably 0.3 to 70 mass%, even more preferably 0.5 to 50 mass%, and particularly preferably 1 to 30 mass%, relative to the total amount of the composition.

[0064] When oleanolic acid and its salts are contained, the content of oleanolic acid and its salts alone is not particularly limited, but is, for example, preferably 0.1 to 95 mass%, more preferably 1 to 80 mass%, even more preferably 10 to 75 mass%, and particularly preferably 20 to 70 mass%, relative to the total amount of the composition.

[0065] When lactoferrin is contained, the content of lactoferrin alone is not particularly limited, but is, for example, preferably 0.1 to 95 mass% relative to the total amount of the composition, more preferably 1 to 80 mass%, even more preferably 10 to 75 mass%, and particularly preferably 20 to 70 mass%.

[0066] When thiamine is contained, the content of thiamine alone is not particularly limited, but is, for example, preferably 0.001 to 95 mass%, more preferably 0.01 to 50 mass%, and even more preferably 0.1 to 10 mass%, relative to the total amount of the composition.

[0067] When niacin is contained, the content of niacin alone is not particularly limited, but is, for example, preferably 0.01 to 95 mass%, more preferably 0.1 to 80 mass%, and even more preferably 1 to 75 mass%, relative to the total amount of the composition.

[0068] When inositol is contained, the content of inositol alone is not particularly limited, but is, for example, preferably 1 to 95 mass%, more preferably 5 to 80 mass%, and even more preferably 10 to 75 mass%, relative to the total amount of the composition.

[0069] When phytic acid is contained, the content of phytic acid alone is not particularly limited, but is, for example, preferably 1 to 95 mass %, more preferably 10 to 80 mass %, and even more preferably 20 to 75 mass %, relative to the total amount of the composition.

[0070] When pantothenic acid is contained, the content of pantothenic acid alone is not particularly limited, but is, for example, preferably 0.001 to 95 mass%, more preferably 0.01 to 50 mass%, and even more preferably 0.1 to 10 mass%, relative to the total amount of the composition.

[0071] When biotin is contained, the content of biotin alone is not particularly limited, but is, for example, preferably 0.00005 to 50 mass%, more preferably 0.0001 to 10 mass%, and even more preferably 0.001 to 1 mass%, relative to the total amount of the composition.

[0072] When folic acid is contained, the content of folic acid alone is not particularly limited, but is, for example, preferably 0.00005 to 50 mass%, more preferably 0.0001 to 10 mass%, even more preferably 0.001 to 1 mass%, and particularly preferably 20 to 70 mass%, relative to the total amount of the composition.

[0073] [Application] The HIF-suppressing composition of the present invention is also suitable for use in improving conditions, symptoms, and diseases associated with the inhibition of HIF. As described above, Non-Patent Document 2 and other publications show the association between HIF and ocular abnormalities. Furthermore, Non-Patent Document 3 and other publications show that abnormally high levels of HIF are observed in retinal pigment epithelial cells (RPE cells) with aging. Therefore, inhibiting HIF in retinal cells is also effective for anti-aging of the eye.

[0074] In addition, it is known that the expression of VEGF is regulated by HIF-1α and HIF-2α, which are types of HIF. Therefore, by inhibiting HIF in retinal cells, it is effective to suppress VEGF in the eye and angiogenesis induced by VEGF. Thus, by inhibiting HIF in retinal cells, angiogenesis is suppressed, so it is also effective to maintain the homeostasis of vascular tissue in the eye.

[0075] In addition, it is known that the inhibition of HIF has a therapeutic effect on retinal ganglion cells and photoreceptor cell disorders in the posterior segment of the eye, and a protective effect on retinal ganglion cells and photoreceptor cells. Therefore, the above-mentioned components are effective against posterior segment diseases and abnormalities in the posterior segment of the eye, including glaucoma and age-related macular degeneration.

[0076] In this specification, the posterior segment of the eye refers to the area inside the back surface of the crystalline lens of the eye, and includes the retina, macula, sclera, choroid, vitreous body, optic nerve, etc. Among the areas of the posterior segment of the eye, it is preferable to target the retina from the viewpoint of significantly exhibiting the effects of the present invention.

[0077] In this specification, the term "posterior segment abnormality" refers to a state in which some abnormality or finding occurs in at least one selected from the group consisting of the retina, macula, sclera, choroid, vitreous body, and optic nerve, although it does not correspond to a disease in the posterior segment. Abnormalities and (clinical) findings in the posterior segment can be found, for example, through a health check, a medical examination by a doctor, a clinical test, etc., but are not limited thereto. Examples of tests for the posterior segment include fundus examination, intraocular pressure test, fundus three-dimensional image analysis (OCT) test, visual field test, refraction test, slit lamp microscopy test, etc. Among these tests, for example, in fundus examination, abnormal findings in the posterior segment are evaluated from the viewpoint of hypertensive changes and arteriosclerotic changes using "Scheie's classification". In addition, in addition to the "Scheie classification", fundus examination may show findings such as enlarged optic nidus, leopard-striated fundus, hard exudates, drusen, suspected glaucoma, hemorrhage, suspected hemorrhage, retinal choroidal atrophy, suspected cataract, crossing phenomenon, exudates, retinal degeneration, meandering, suspected macular degeneration, conus, optic disc findings, etc. The present invention may also be used for reducing the risk, preventing, or treating posterior segment abnormalities.

[0078] As used herein, prevention of posterior ocular abnormalities refers to preventing or delaying the onset of posterior ocular abnormalities, or reducing the risk of onset of posterior ocular abnormalities.

[0079] As used herein, improvement of posterior ocular abnormalities refers to alleviating or improving a condition in which there is an abnormality in the posterior ocular segment, preventing or delaying the worsening of a condition in which there is an abnormality, or preventing, delaying, or reversing the progression of a condition in which there is an abnormality in the posterior ocular segment.

[0080] The posterior ocular disease targeted by the composition for reducing, preventing or treating the risk of posterior ocular disease of the present invention is not limited, but includes at least one selected from the group consisting of age-related macular degeneration, diabetic retinopathy, diabetic macular edema, retinitis pigmentosa, proliferative vitreous retinopathy, retinal artery occlusion, retinal vein occlusion, uveitis, Leber's disease, retinopathy of prematurity, retinal detachment, retinal pigment epithelial detachment, central serous chorioretinopathy, central exudative chorioretinopathy, polypoidal choroidal vasculopathy, multifocal choroiditis, neovascular maculopathy, retinal aneurysm, retinal angiomatous proliferation, optic nerve disorders caused by these diseases, optic nerve disorders caused by glaucoma and ischemic optic nerve disorders. Among these posterior ocular diseases, it is preferable to target age-related macular degeneration and / or retinitis pigmentosa from the viewpoint of significantly exerting the effects of the present invention.

[0081] As used herein, prevention of a posterior ocular disease refers to preventing or delaying the onset of a posterior ocular disease or symptom, or reducing the risk of onset of a posterior ocular disease or symptom.

[0082] As used herein, treatment of a posterior ocular disease refers to alleviating or improving a posterior ocular disease or symptom, preventing or slowing the worsening of a disease or symptom, or preventing, slowing, or reversing the progression of a posterior ocular disease or symptom.

[0083] Improvement of posterior segment abnormalities or prevention of posterior segment diseases includes, but is not limited to, reducing the risk of developing at least one selected from the group consisting of age-related macular degeneration, diabetic retinopathy, diabetic macular edema, retinitis pigmentosa, proliferative vitreous retinopathy, retinal artery occlusion, retinal vein occlusion, uveitis, Leber's disease, retinopathy of prematurity, retinal detachment, retinal pigment epithelial detachment, central serous chorioretinopathy, central exudative chorioretinopathy, polypoidal choroidal vasculopathy, multifocal choroiditis, neovascular maculopathy, retinal aneurysm, retinal angiomatous proliferation, optic neuropathy caused by these diseases, optic neuropathy caused by glaucoma, and ischemic optic neuropathy.

[0084] The composition of the present invention can be added to or mixed with food, beverage, feed, or pet food. Alternatively, it can be used as a beverage or food as it is. Alternatively, it can be added to or mixed with food and beverage products labeled as having functional properties such as retinal protection, anti-aging of the eye, VEGF inhibition, angiogenesis inhibition, maintenance of homeostasis of vascular tissue, and risk reduction or prevention of posterior eye disease, i.e., health foods, functional foods, foods for the sick, and foods for specified health uses.

[0085] Health foods, functional foods, foods for the sick, and foods for specified health uses can be used in various formulation forms, specifically, solid preparations (tablets, granules, fine granules, powders, capsules, chewable tablets, etc.), liquid preparations (syrups, suspensions), liquid diets, etc. Foods in formulation form can be produced in the same manner as known pharmaceutical preparations, and can be produced by mixing an active ingredient with a carrier acceptable for use as a food, such as a suitable excipient, and then using conventional means.

[0086] For example, tablets can be prepared by mixing a powdered active ingredient with a pharma- ceutically acceptable carrier component (such as an excipient) and compressing the mixture, and confectionery tablets such as candy can be prepared by pouring the mixture into a mold. Tablets can also be sugar-coated to form sugar-coated tablets. Furthermore, tablets can be single-layer tablets or laminated tablets such as double-layer tablets.

[0087] Powdered granules such as granules may be prepared by various granulation methods (such as extrusion granulation, grinding granulation, dry compaction granulation, fluidized bed granulation, rolling granulation, and high-speed stirring granulation), and tablets can be prepared by an appropriate combination of the above-mentioned granulation methods and tableting methods (wet tableting, direct tableting), etc.

[0088] Capsules can be prepared by filling a capsule (soft or hard capsule) with a powdered agent (powder, granules, etc.) by a conventional method.

[0089] The liquid preparation can be prepared by dissolving or dispersing each component in an aqueous medium (purified water, purified water containing ethanol, etc.) which is a carrier component, filtering or sterilizing the mixture as necessary, filling the mixture into a predetermined container, and sterilizing the mixture. The preferred dosage form of the solid preparation of the present invention is a capsule or tablet, and a soft capsule (soft capsule) is more preferred.

[0090] Soft capsules have a smooth surface and are easy to swallow, and are therefore preferred by users. Typical methods for producing soft capsules include the plate method, the rotary method, and the seamless method.

[0091] In the rotary method (punching method), a sheet-like capsule shell sandwiches the flowing filling contents and forms a capsule shape along the holes of a rotating cylindrical die. On the other hand, in the seamless method (dropping method), the capsule shell composition and contents are simultaneously discharged from multiple concentric nozzles to form a seamless capsule shape.

[0092] The base material for the soft capsule shell is not particularly limited, but starch, pullulan, cellulose, polyvinyl alcohol, gelatin, succinated gelatin, etc. can be used, with starch, gelatin, and succinated gelatin being preferred, and gelatin and succinated gelatin being more preferred. These may be used alone or in combination of two or more kinds.

[0093] The composition of the present invention can also be manufactured as liquid beverages such as soups, juices, fruit juice drinks, milk, dairy drinks, whey drinks, lactic acid bacteria drinks, tea drinks, alcoholic drinks, coffee drinks, carbonated drinks, soft drinks, water drinks, cocoa drinks, jelly drinks, sports drinks, and diet drinks; semi-solid foods such as pudding and yogurt; noodles, confectioneries, spreads, and the like.

[0094] When the composition of the present invention is prepared as a food composition, various food additives may be blended therein, such as antioxidants, colorants, flavorings, seasonings, sweeteners, acidulants, pH adjusters, quality stabilizers, preservatives, etc.

[0095] When the composition of the present invention is prepared as a pharmaceutical composition, it is prepared as a formulation containing at least one active ingredient selected from the group consisting of hydroxycitric acid and its salt, Garcinia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salt, and oleanolic acid and its salt, and preferably a pharmaceutical acceptable carrier.The pharmaceutical acceptable carrier generally refers to an inactive, non-toxic, solid or liquid filler, diluent, or encapsulating material that does not react with the active ingredient, such as water, ethanol, polyols, suitable mixtures thereof, and solvents or dispersion media such as vegetable oils.

[0096] The pharmaceutical composition is administered orally or parenterally, for example, into the oral cavity, into the digestive tract, or into the nasal cavity. Oral preparations include solid preparations (tablets, granules, fine granules, powders, capsules, chewable tablets, etc.) and liquid preparations (syrups, suspensions, inhalants, etc.). Parenteral preparations include eye drops, drops, nasal drops, injections, etc.

[0097] The pharmaceutical composition may further contain additives commonly used in the pharmaceutical field. Such additives include, for example, excipients, binders, disintegrants, lubricants, antioxidants, colorants, flavoring agents, etc., and can be used appropriately as needed. In order to achieve sustained release so as to act for a long time, the composition may also be coated with a known retardant, etc. The pharmaceutical composition may further contain other additives or drugs, such as antacids and gastric mucosa protectants, as needed.

[0098] The pharmaceutical composition can be applied in the form of an oral composition, an internally ingested composition, etc. The pharmaceutical composition may be used therapeutically or non-therapeutically.

[0099] The oral intake or dosage of hydroxycitric acid and / or its salts per day for adults can be appropriately determined according to individual condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The oral intake or dosage of hydroxycitric acid and / or its salts per day for adults is, for example, preferably 0.01 mg / kg body weight / day or more, more preferably 0.1 mg / kg body weight / day or more, even more preferably 0.5 mg / kg body weight / day or more, particularly preferably 1 mg / kg body weight / day or more, and most preferably 10 mg / kg body weight / day or more. The oral intake or dosage of hydroxycitric acid and / or its salts per day for adults is, for example, preferably 400 mg / kg body weight / day or less, more preferably 300 mg / kg body weight / day or less, even more preferably 100 mg / kg body weight / day or less, particularly preferably 50 mg / kg body weight / day or less, and most preferably 30 mg / kg body weight / day or less. The oral intake or dosage of hydroxycitric acid and / or its salt per day for an adult is, for example, preferably 0.01-400 mg / kg body weight / day, more preferably 0.1-300 mg / kg body weight / day, even more preferably 0.5-100 mg / kg body weight / day, particularly preferably 1-50 mg / kg body weight / day, and most preferably 10-30 mg / kg body weight / day. The content of hydroxycitric acid and / or its salt can be the amount that results in the above intake or dosage. The oral intake or dosage per day for an adult may be divided into 1-6 capsules, 1-4 capsules, 1-3 capsules, or 1-2 capsules depending on the dosage form, for example, in the case of capsules.

[0100] The daily oral intake or dosage of Garcinia and / or its extract for an adult can be appropriately determined depending on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The daily oral intake or dosage of Garcinia and / or its extract for an adult is, for example, preferably 0.02 mg / kg body weight / day or more, more preferably 0.2 mg / kg body weight / day or more, even more preferably 1.0 mg / kg body weight / day or more, particularly preferably 2 mg / kg body weight / day or more, and most preferably 20 mg / kg body weight / day or more. The daily oral intake or dosage of Garcinia and / or its extract for an adult is, for example, preferably 800 mg / kg body weight / day or less, more preferably 600 mg / kg body weight / day or less, even more preferably 200 mg / kg body weight / day or less, particularly preferably 100 mg / kg body weight / day or less, and most preferably 60 mg / kg body weight / day or less. The oral intake or dosage of Garcinia and / or its extract per day for an adult is, for example, preferably 0.02 to 800 mg / kg body weight / day, more preferably 0.2 to 600 mg / kg body weight / day, even more preferably 1.0 to 200 mg / kg body weight / day, particularly preferably 2 to 100 mg / kg body weight / day, and most preferably 20 to 60 mg / kg body weight / day. The content of Garcinia and / or its extract can be the amount that results in the above intake or dosage. The oral intake or dosage per day for an adult may be divided into 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules depending on the dosage form, for example, in the case of capsules.

[0101] The oral intake or dosage of lactoferrin for an adult per day can be appropriately determined according to individual condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors.The oral intake or dosage of lactoferrin for an adult per day is, for example, preferably 0.01 mg / kg body weight / day or more, more preferably 0.1 mg / kg body weight / day or more, even more preferably 0.5 mg / kg body weight / day or more, particularly preferably 1 mg / kg body weight / day or more, and most preferably 2 mg / kg body weight / day or more.The oral intake or dosage of lactoferrin for an adult per day is, for example, preferably 200 mg / kg body weight / day or less, more preferably 150 mg / kg body weight / day or less, even more preferably 100 mg / kg body weight / day or less, particularly preferably 50 mg / kg body weight / day or less, and most preferably 20 mg / kg body weight / day or less. The oral intake or dosage of lactoferrin per day for an adult is, for example, preferably 0.01-200 mg / kg body weight / day, more preferably 0.1-150 mg / kg body weight / day, even more preferably 0.5-100 mg / kg body weight / day, particularly preferably 1-50 mg / kg body weight / day, and most preferably 2-20 mg / kg body weight / day. The lactoferrin content can be the amount that results in the above intake or dosage. The oral intake or dosage per day for an adult may be divided into 1-6 capsules, 1-4 capsules, 1-3 capsules, or 1-2 capsules depending on the dosage form, for example, in the case of capsules.

[0102] The daily oral intake or dosage of carrot and / or its extract for an adult can be appropriately determined depending on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The daily oral intake or dosage of carrot and / or its extract for an adult is, for example, preferably 0.02 mg / kg body weight / day or more, more preferably 0.2 mg / kg body weight / day or more, even more preferably 1.0 mg / kg body weight / day or more, particularly preferably 2 mg / kg body weight / day or more, and most preferably 20 mg / kg body weight / day or more. The daily oral intake or dosage of carrot and / or its extract for an adult is, for example, preferably 800 mg / kg body weight / day or less, more preferably 600 mg / kg body weight / day or less, even more preferably 200 mg / kg body weight / day or less, particularly preferably 100 mg / kg body weight / day or less, and most preferably 60 mg / kg body weight / day or less. The oral intake or dosage of ginseng and / or its extract per day for an adult is, for example, preferably 0.02 to 800 mg / kg body weight / day, more preferably 0.2 to 600 mg / kg body weight / day, even more preferably 1.0 to 200 mg / kg body weight / day, particularly preferably 2 to 100 mg / kg body weight / day, and most preferably 20 to 60 mg / kg body weight / day. The content of ginseng and / or its extract can be the amount that results in the above intake or dosage. The oral intake or dosage per day for an adult may be divided into 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules depending on the dosage form, for example, in the case of capsules.

[0103] The oral intake or dosage of pyridoxine and / or its salts per day for adults can be appropriately determined according to individual condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The oral intake or dosage of pyridoxine and / or its salts per day for adults is, for example, preferably 0.0001 mg / kg body weight / day or more, more preferably 0.001 mg / kg body weight / day or more, even more preferably 0.005 mg / kg body weight / day or more, particularly preferably 0.01 mg / kg body weight / day or more, and most preferably 0.02 mg / kg body weight / day or more. The oral intake or dosage of pyridoxine and / or its salts per day for adults is, for example, preferably 50 mg / kg body weight / day or less, more preferably 10 mg / kg body weight / day or less, even more preferably 5 mg / kg body weight / day or less, particularly preferably 1 mg / kg body weight / day or less, and most preferably 0.5 mg / kg body weight / day or less. The oral intake or dosage of pyridoxine and / or its salt per day for an adult is, for example, preferably 0.0001-50 mg / kg body weight / day, more preferably 0.001-10 mg / kg body weight / day, even more preferably 0.005-5 mg / kg body weight / day, particularly preferably 0.01-1 mg / kg body weight / day, and most preferably 0.02-0.5 mg / kg body weight / day. The content of pyridoxine and / or its salt can be the amount that results in the above intake or dosage. The oral intake or dosage per day for an adult may be divided into 1-6 capsules, 1-4 capsules, 1-3 capsules, or 1-2 capsules depending on the dosage form, for example, in the case of capsules.

[0104] The oral intake or dosage of oleanolic acid and / or its salts per day for an adult can be appropriately determined depending on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The oral intake or dosage of oleanolic acid and / or its salts per day for an adult is, for example, preferably 0.01 mg / kg body weight / day or more, more preferably 0.1 mg / kg body weight / day or more, even more preferably 0.5 mg / kg body weight / day or more, particularly preferably 1 mg / kg body weight / day or more, and most preferably 10 mg / kg body weight / day or more. The oral intake or dosage of oleanolic acid and / or its salts per day for an adult is, for example, preferably 2000 mg / kg body weight / day or less, more preferably 1500 mg / kg body weight / day or less, even more preferably 1000 mg / kg body weight / day or less, particularly preferably 800 mg / kg body weight / day or less, and most preferably 500 mg / kg body weight / day or less. The oral intake or dosage of oleanolic acid and / or its salt per day for an adult is, for example, preferably 0.01-2000 mg / kg body weight / day, more preferably 0.1-1500 mg / kg body weight / day, even more preferably 0.5-1000 mg / kg body weight / day, particularly preferably 1-800 mg / kg body weight / day, and most preferably 10-500 mg / kg body weight / day. The content of oleanolic acid and / or its salt can be the amount that results in the above intake or dosage. The oral intake or dosage per day for an adult may be divided into 1-6 capsules, 1-4 capsules, 1-3 capsules, or 1-2 capsules depending on the dosage form, for example, in the case of capsules.

[0105] The daily oral intake or dosage of thiamine for an adult can be appropriately determined depending on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The daily oral intake or dosage of thiamine for an adult is, for example, preferably 0.0001 mg / kg body weight / day or more, more preferably 0.001 mg / kg body weight / day or more, and even more preferably 0.005 mg / kg body weight / day or more. The daily oral intake or dosage of thiamine for an adult is, for example, preferably 10 mg / kg body weight / day or less, more preferably 1 mg / kg body weight / day or less, and even more preferably 0.1 mg / kg body weight / day or less. The daily oral intake or dosage of thiamine for an adult is, for example, preferably 0.0001 to 10 mg / kg body weight / day, more preferably 0.001 to 1 mg / kg body weight / day, and even more preferably 0.005 to 0.1 mg / kg body weight / day. The amount of thiamine contained can be the amount that results in the above-mentioned intake or dosage. The oral intake or dosage for an adult per day can be divided into 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules depending on the dosage form.

[0106] The oral intake or dosage of niacin per day for an adult can be appropriately determined according to individual condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors.The oral intake or dosage of niacin per day for an adult is, for example, preferably 0.01 mg / kg body weight / day or more, more preferably 0.1 mg / kg body weight / day or more, and even more preferably 0.5 mg / kg body weight / day or more, as nicotinic acid.The oral intake or dosage of niacin per day for an adult is, for example, preferably 100 mg / kg body weight / day or less, more preferably 50 mg / kg body weight / day or less, and even more preferably 10 mg / kg body weight / day or less, as nicotinic acid. The oral intake or dosage of niacin per day for an adult is, for example, preferably 0.01-100 mg / kg body weight / day, more preferably 0.1-50 mg / kg body weight / day, and even more preferably 0.5-10 mg / kg body weight / day. The content of niacin can be the amount that results in the above intake or dosage. The oral intake or dosage per day for an adult may be divided into 1-6 capsules, 1-4 capsules, 1-3 capsules, or 1-2 capsules depending on the dosage form, for example, in the case of capsules.

[0107] The daily oral intake or dosage of inositol for an adult can be appropriately determined according to the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors.The daily oral intake or dosage of inositol for an adult is, for example, preferably 0.01 mg / kg body weight / day or more, more preferably 0.1 mg / kg body weight / day or more, and even more preferably 0.5 mg / kg body weight / day or more.The daily oral intake or dosage of inositol for an adult is, for example, preferably 200 mg / kg body weight / day or less, more preferably 150 mg / kg body weight / day or less, and even more preferably 100 mg / kg body weight / day or less. The oral intake or dosage of inositol per day for an adult is, for example, preferably 0.01-200 mg / kg body weight / day, more preferably 0.1-150 mg / kg body weight / day, even more preferably 0.5-100 mg / kg body weight / day, particularly preferably 1-50 mg / kg body weight / day, and most preferably 2-20 mg / kg body weight / day. The content of inositol can be the amount that results in the above intake or dosage. The oral intake or dosage per day for an adult may be divided into 1-6 capsules, 1-4 capsules, 1-3 capsules, or 1-2 capsules depending on the dosage form, for example, in the case of capsules.

[0108] The oral intake or dosage of phytic acid per day for an adult can be appropriately determined depending on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The oral intake or dosage of phytic acid per day for an adult is, for example, preferably 0.01 mg / kg body weight / day or more, more preferably 0.1 mg / kg body weight / day or more, and even more preferably 1 mg / kg body weight / day or more. The oral intake or dosage of phytic acid per day for an adult is, for example, preferably 200 mg / kg body weight / day or less, more preferably 150 mg / kg body weight / day or less, and even more preferably 100 mg / kg body weight / day or less. The oral intake or dosage of phytic acid per day for an adult is, for example, preferably 0.01 to 200 mg / kg body weight / day, more preferably 0.1 to 150 mg / kg body weight / day, even more preferably 0.5 to 100 mg / kg body weight / day, particularly preferably 1 to 50 mg / kg body weight / day, and most preferably 2 to 20 mg / kg body weight / day. The content of phytic acid can be the amount that results in the above intake or dosage. The oral intake or dosage per day for an adult may be divided into 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules depending on the dosage form, for example, in the case of capsules.

[0109] The daily oral intake or dosage of pantothenic acid for an adult can be appropriately determined depending on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The daily oral intake or dosage of pantothenic acid for an adult is, for example, preferably 0.001 mg / kg body weight / day or more, more preferably 0.005 mg / kg body weight / day or more, and even more preferably 0.01 mg / kg body weight / day or more. The daily oral intake or dosage of pantothenic acid for an adult is, for example, preferably 100 mg / kg body weight / day or less, more preferably 10 mg / kg body weight / day or less, and even more preferably 1 mg / kg body weight / day or less. The daily oral intake or dosage of pantothenic acid for an adult is, for example, preferably 0.001 to 100 mg / kg body weight / day, more preferably 0.005 to 10 mg / kg body weight / day, and even more preferably 0.01 to 1 mg / kg body weight / day. The content of pantothenic acid can be the amount that results in the above intake or dosage. The daily oral intake or dosage for an adult may be divided into 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules depending on the dosage form, for example, in the case of capsules.

[0110] The oral intake or dosage of biotin per day for an adult can be appropriately determined depending on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The oral intake or dosage of biotin per day for an adult is, for example, preferably 0.01 mg / kg body weight / day or more, more preferably 0.1 mg / kg body weight / day or more, and even more preferably 0.5 mg / kg body weight / day or more. The oral intake or dosage of biotin per day for an adult is, for example, preferably 100 mg / kg body weight / day or less, more preferably 50 mg / kg body weight / day or less, and even more preferably 10 mg / kg body weight / day or less. The oral intake or dosage of biotin per day for an adult is, for example, preferably 0.01 to 100 mg / kg body weight / day, more preferably 0.1 to 50 mg / kg body weight / day, and even more preferably 0.5 to 10 mg / kg body weight / day. The amount of biotin contained can be the amount that results in the above-mentioned intake or administration amount. The oral intake or administration amount per day for an adult may be divided into 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules depending on the dosage form, for example, in the case of capsules.

[0111] The oral intake or dosage of folic acid per day for an adult can be appropriately determined depending on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The oral intake or dosage of folic acid per day for an adult is, for example, preferably 0.01 μg / kg body weight / day or more, more preferably 0.1 μg / kg body weight / day or more, and even more preferably 0.5 μg / kg body weight / day or more. The oral intake or dosage of folic acid per day for an adult is, for example, preferably 200 μg / kg body weight / day or less, more preferably 150 μg / kg body weight / day or less, and even more preferably 100 μg / kg body weight / day or less. The oral intake or dosage of folic acid per day for an adult is, for example, preferably 0.01 to 200 mg / kg body weight / day, more preferably 0.1 to 150 μg / kg body weight / day, and even more preferably 0.5 to 100 μg / kg body weight / day. The content of folic acid can be the amount that results in the above-mentioned intake or dosage. The oral intake or dosage per day for adults may be divided into 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules depending on the dosage form.

[0112] The oral intake or administration amount of at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine, niacin, inositol, phytic acid, pantothenic acid, biotin, and folic acid can be appropriately determined according to the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The oral intake or administration amount of the above-mentioned components per time is, for example, preferably 0.01 mg or more, more preferably 0.1 mg or more, even more preferably 0.5 mg or more, particularly preferably 1 mg or more, and most preferably 10 mg or more. The oral intake or administration amount of the above-mentioned components per time is, for example, preferably 500 mg or less, more preferably 400 mg or less, even more preferably 300 mg or less, particularly preferably 200 mg or less, and most preferably 100 mg or less. The oral intake or administration amount of the above-mentioned component per time is, for example, preferably 0.01 to 500 mg, more preferably 0.1 to 400 mg, further preferably 0.5 to 300 mg, particularly preferably 1 to 200 mg, and most preferably 10 to 100 mg.

[0113] The food composition for preventing and / or improving posterior ocular abnormalities, or the agent for preventing and / or treating posterior ocular diseases of the present invention can be ingested or administered once or several times a day, typically 1 to 6 times a day, 1 to 3 times a day, 1 to 2 times a day, or at any period and interval.

[0114] The present invention may also have the following aspects. A composition for suppressing HIF, comprising at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; A composition for retinal protection comprising at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; an anti-aging composition for the eye, comprising at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; A composition for suppressing VEGF in the eye, comprising at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; A composition for inhibiting ocular neovascularization, comprising at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; A composition for maintaining homeostasis of ocular vascular tissue, comprising at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; A composition for reducing the risk of, preventing or treating a posterior ocular disease, comprising at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid and rice bran; Use of a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran, for the production of a HIF suppressor; Use of a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran, for the manufacture of a retinal protective agent; Use of a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran, for the manufacture of an anti-aging agent for the eye; Use of a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran, for the manufacture of an ocular VEGF inhibitor; Use of a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran, for the production of an ocular angiogenesis inhibitor; Use of a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran, for the manufacture of an agent for maintaining homeostasis of ocular vascular tissue; Use of a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran, for the manufacture of an agent for reducing the risk of, preventing, or treating a posterior ocular disease; A method for suppressing HIF, comprising having a person ingest a composition containing at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; A method for protecting the retina, comprising having a human ingest a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; an anti-aging method for the eye, comprising having a person ingest a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; A method for suppressing ocular VEGF, comprising having a human ingest a composition containing at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; A method for inhibiting ocular neovascularization, comprising having a human ingest a composition containing at least one member selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; A method for maintaining homeostasis of ocular vascular tissue, comprising having a human ingest a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid, and rice bran; A method for reducing the risk of, preventing or treating a posterior ocular disease, comprising having a person ingest a composition containing at least one selected from the group consisting of hydroxycitric acid and its salts, Garcinia cambogia and its extract, lactoferrin, carrot and its extract, pyridoxine and its salts, oleanolic acid and its salts, thiamine and its salts, niacin, inositol, phytic acid and its salts, pantothenic acid and its salts, biotin, folic acid and rice bran; the composition, use, or method, wherein the posterior ocular disease is at least one selected from the group consisting of age-related macular degeneration, diabetic retinopathy, diabetic macular edema, retinitis pigmentosa, proliferative vitreous retinopathy, retinal artery occlusion, retinal vein occlusion, uveitis, Leber's disease, retinopathy of prematurity, retinal detachment, retinal pigment epithelium detachment, central serous chorioretinopathy, central exudative chorioretinopathy, polypoidal choroidal vasculopathy, multifocal choroiditis, neovascular maculopathy, retinal aneurysm, retinal angiomatous proliferation, optic neuropathy caused by these diseases, optic neuropathy caused by glaucoma, and ischemic optic neuropathy; the composition, use or method, wherein the salt of hydroxycitric acid is a calcium salt; the composition, use or method, wherein the salt of pyridoxine is a hydrochloride or a phosphate; the composition, use or method, wherein the rice bran is defatted rice bran; the composition, use, or method, wherein said niacin is niacinamide; the composition, use or method, wherein the salt of pantothenic acid is a calcium salt; the composition, use, or method, wherein the total content of at least one selected from the group consisting of hydroxycitric acid and salts thereof, Garcinia and extract thereof, lactoferrin, carrot and extract thereof, pyridoxine and salts thereof, oleanolic acid and salts thereof, thiamine and salts thereof, niacin, inositol, phytic acid and salts thereof, pantothenic acid and salts thereof, biotin, folic acid, and rice bran is 0.1 to 95% by mass; the composition, use, or method, wherein the daily oral intake of the hydroxycitric acid and its salts for an adult is 0.01 to 400 mg / kg body weight / day; the composition, use or method, wherein the daily oral intake amount of the Garcinia cambogia and / or an extract thereof per adult is 0.02 to 800 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake of the ginseng and / or an extract thereof for an adult is 0.02 to 800 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake of the pyridoxine and / or a salt thereof per adult is 0.0001 to 50 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake of the oleanolic acid and / or a salt thereof for an adult is 0.01 to 2000 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake of lactoferrin for an adult is 0.01 to 200 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake of thiamine per adult is 0.0001 to 10 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake of niacin per adult is 0.01 to 100 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake amount of the inositol per adult is 0.01 to 200 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake of the phytic acid and / or a salt thereof per adult is 0.01 to 200 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake of the pantothenic acid and / or a salt thereof per adult is 0.001 to 100 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake of biotin per adult is 0.01 to 100 mg / kg body weight / day; the composition, use, or method, wherein the daily oral intake of the folic acid for an adult is 0.01 to 200 mg / kg body weight / day; the composition, use, or method, wherein the amount of the component taken orally per dose is 0.01 to 500 mg; The composition, use or method as described above, wherein the ingredient is taken 1 to 6 times a day; EXAMPLES

[0115] Next, the present invention will be specifically described with reference to examples, but the present invention is not limited to the following examples. Furthermore, the amount of extract shown in the examples is the amount calculated as a dry solid content and including excipients, unless otherwise specified.

[0116] [Test Example 1: Screening of HIF-inhibiting substances 1] We used lentivirus to transduce both HIF-dependent Firefly-Luciferase and the endogenous control CMV-Renilla-Luciferase into a mouse retinal cone cell line (661W) to generate a stable expression line, RH661W. We then treated this line with cobalt chloride (CoCl ), a prolyl hydroxylase (PHD) inhibitor, to generate a stable expression line, RH661W. 2 The addition of luciferin stabilizes HIF, and the addition of luciferin produces a luminescence signal that correlates with HIF activity. Various samples (candidate substances) were added to this RH661W, and the HIF inhibitory effect of the candidate substances on cobalt chloride-induced HIF activity was confirmed.

[0117] So, we put 0.8 × 10 RH661W into a 96-well plate. 4 After 24 hours, the cells were attached to the bottom of the plate, and then 200 μM CoCl 2 The HIF activity was induced by adding 1 mg / ml of each candidate substance. The candidate substances were (-)-hydroxycitric acid calcium salt, Garcinia cambogia extract (Bioactives Japan Co., Ltd.), ginseng extract (Matsuura Pharmaceutical Co., Ltd.), pyridoxine hydrochloride, and 20-23% vitamin B 6 , defatted rice bran (Oryza Oil & Fat Chemical Co., Ltd.), oleanolic acid, and lactoferrin were used. After 24 hours, the luminescence intensity was measured using the Promega Dual-Glo Luciferase assay system to verify the HIF inhibitory effect.

[0118] The activity of each was calculated as follows. The number of photons emitted by the cells was measured using a plate reader (BioTek, SYNERGY HTX), the ratio was calculated, and the average value of each candidate substance was calculated. The ratio when the candidate substance was added was divided by the ratio when only cobalt chloride was added, and the average value of cell + CoCl 2 The ratio was calculated, and the results are shown in Table 1.

[0119] [Test Example 2: Screening of HIF-inhibiting substances 2] We used lentivirus to transduce both HIF-dependent Firefly-luciferase and the endogenous control CMV-Renilla-luciferase into human retinal pigment epithelial cell line (ARPE) to generate a stable expression line RHARPE. We then treated this RHARPE with cobalt chloride (CoCl ), a prolyl hydroxylase (PHD) inhibitor, to generate a stable expression line RHARPE. 2 By adding luciferin, HIF can be stabilized, and a luminescence signal that correlates with HIF activity can be obtained by adding various samples (candidate substances) to this RHARPE, and the HIF inhibitory effect of the candidate substances on cobalt chloride-induced HIF activity was confirmed.

[0120] So, 0.8 × 10 RHARPE was added to a 96-well plate. 4 After 24 hours, the cells were attached to the bottom of the plate, and then 200 μM CoCl 2 The candidate substances were added at 1 mg / ml. The candidate substances were ginseng extract (Matsuura Pharmaceutical Co., Ltd.), Garcinia cambogia extract (BioActives Japan Co., Ltd.), (-)-hydroxycitric acid calcium salt, lactoferrin, and vitamin B 1 , Vitamin B 6 , niacinamide (nicotinamide), inositol, phytic acid, calcium D-pantothenate, d-biotin, folic acid, defatted rice bran (Oryza Oil & Fat Chemical Co., Ltd.), and oleanolic acid were used. After 24 hours, the luminescence intensity was measured using the Promega Dual-Glo Luciferase assay system to verify the HIF inhibitory effect.

[0121] The activity of each was calculated as follows. The number of photons emitted by the cells was measured using a plate reader (BioTek, SYNERGY HTX), the ratio was calculated, and the average value of each candidate substance was calculated. The ratio when the candidate substance was added was divided by the ratio when only cobalt chloride was added, and the average value of cell + CoCl 2 The ratio was calculated, and the results are shown in Table 2.

[0122] [Table 1]

[0123] [Table 2]

[0124] As shown in Tables 1 and 2, HIF inhibitory activity was observed for hydroxycitric acid and its salts, Garcinia and its extract, carrot and its extract, pyridoxine and its salts, oleanolic acid, lactoferrin, thiamine, niacin, inositol, phytic acid, pantothenic acid, biotin, defatted rice bran, and folic acid.

[0125] It is known that abnormally high levels of HIF are observed in retinal pigment epithelial cells (RPE cells) with age. In Test Example 1, retinal cone cells, which also play an important role as photoreceptor cells in the retina, were used to screen for HIF inhibitors, and in Test Example 2, retinal pigment epithelial cells were used to screen for HIF inhibitors. The above ingredients that were confirmed to suppress HIF are capable of inhibiting HIF in retinal cells, suggesting that they are effective for anti-aging of the eye.

[0126] In addition, it is known that the expression of VEGF is regulated by HIF-1α and HIF-2α, which are types of HIF. Therefore, inhibiting HIF in retinal cells is effective in suppressing VEGF in the eye and angiogenesis induced by VEGF. Thus, it was suggested that inhibiting HIF in retinal cells is also effective in maintaining the homeostasis of ocular vascular tissue because angiogenesis is suppressed by inhibiting HIF in retinal cells.

[0127] In addition, it is known that inhibiting HIF has a therapeutic effect on retinal ganglion cells and photoreceptor cell disorders in the posterior segment of the eye, and a protective effect on retinal ganglion cells and photoreceptor cells. Therefore, it is suggested that the above-mentioned components are effective against posterior segment diseases and abnormalities in the posterior segment of the eye, including glaucoma and age-related macular degeneration.

[0128] [Test Example 3: Electroretinogram (ERG) Test (protection of retinal photoreceptor cells by suppressing the decrease in retinal action potential)] Six-week-old male BALB / cA mice were fed a control diet (MF, Oriental Yeast Co., Ltd.) for the control group, and a diet containing 3g / kg / day of Garcinia cambogia extract (Bioactives Japan Co., Ltd.) mixed with the control diet for a total of 10 weeks for the Garcinia treatment group. On the 8th week of treatment, after dark adaptation from the previous day, the mice were exposed to 3000 Lux light for 1 hour. ERG was measured 1 and 2 weeks after exposure. Eight-week-old BALB / cA mice (male) were intraperitoneally administered corn oil (SIGMA-ALDRICH corn oil 2303205, the same below) to the control group, and (-)-hydroxycitric acid suspended in corn oil at 300 mg / kg / day / 1000 μL to the hydroxycitric acid group, 5 days / week for a total of 2 weeks. In the first week of administration, after dark adaptation from the previous day, the mice were exposed to 3000 Lux light for 1 hour. ERG was measured one week after exposure. For ERG, full-field flash ERG responses were recorded using a Ganzfeld dome, measuring device, and LED stimulator (PuREC, MAYO Corporation, Japan). After overnight dark adaptation, mice were anesthetized with MMB. The relevant electrode was recorded with a contact lens electrode, and the indifferent electrode was placed subcutaneously between the eyes. A clipping electrode to the tail was used as ground. ERG responses were obtained from both eyes of each animal. First, stimulation under dark adaptation (10 cd s / m 2 ) dark-topic responses were recorded. Photopic responses were then recorded. 2 ) under stimulation (20cds / m 2 ) photopic responses were recorded. All mice were kept warm during the procedure using a heat pad. The ERG a-wave was fitted to the following equation: Based on the Lamb and Pugh model, responses (R) were calculated as a function of time (T) and flash intensity (I). Rmax, S, and Td are the maximum amplitude, sensitivity, and delay before the onset of the a-wave, respectively. S and Rmax data were transformed to logarithmic values ​​for data analysis.

[0129] [Formula 1] R(I,T)= {1-exp [-I·S·(T-Td)2]·Rmax

[0130] After being low-pass filtered at 30 Hz, the ERG b-wave was fitted to the Naka-Rushton equation: where R is the amplitude, Rmax is the maximum amplitude, I is the flash intensity, K is the light intensity at half-maximum amplitude, and n is a dimensionless compression constant.

[0131] [Formula 2] R = In / In + Kn Rmax

[0132] Figure 1 (A) to (D) show the dark-adapted stimulation (10 cds / m ) in the Garcinia cambogia group and the control group. 2 2 shows the results of ERG (rod system function) measurement using a Garcinia cambogia-administered group and a control-administered group. 2 3 shows the results of ERG (cone system function) measurement by electroretinogram (ERG). In this specification, the number n in the figures showing the results of electroretinogram (ERG) tests indicates the number of mice evaluated.

[0133] As shown in Figures 1 and 2, the decrease in ERG amplitude was suppressed in the Garcinia administration group compared to the control administration group at both the 1st and 2nd week of irradiation, and the suppressive effect of Garcinia administration on retinal light damage was confirmed. Therefore, the retinal protective effect of Garcinia in the retinal light damage model was confirmed.

[0134] Figure 3 (A)-(B) shows the dark-adapted stimulation (10 cd s / m 2 ) in the hydroxycitric acid-administered group and the corn oil (control)-administered group. 2 ) shows the results of ERG (cone system function) measurement.

[0135] As shown in Figures 3 and 4, the decrease in ERG amplitude was suppressed in the hydroxycitric acid group compared to the corn oil group at 1 week after irradiation, and the suppressive effect of hydroxycitric acid administration on retinal light damage was confirmed. Therefore, the retinal protective effect of hydroxycitric acid in the retinal light damage model was confirmed. Since hydroxycitric acid is an active ingredient contained in large amounts in Garcinia cambogia, it was confirmed that the results of Garcinia administration in Figures 1 and 2 and the results of hydroxycitric acid administration in Figures 3 and 4 are consistent.

[0136] [Test Example 4: Laser CNV Test Method (Angiogenesis Inhibition) 1] The diet was orally administered to 4-week-old C57BL6 / J mice (male) for a total of 49 and 63 days, respectively, and on the 42nd day after the start of administration, a choroidal neovascularization (CNV) model was created by laser irradiation at 200 mW, 0.1 sec, and 75 μm, and the CNV volume was quantified 7 days after irradiation. The control group was administered control diet (MF, Oriental Yeast Co., Ltd.), and the Garcinia administration group was administered diet containing Garcinia cambogia extract (BioActives Japan Co., Ltd.) mixed with the control diet at 300 mg / kg / day.

[0137] Specifically, C57BL / 6 mice were anesthetized with MMB, and 5 sites per eye were irradiated near the optic disc with an argon laser (spot size 75 μm, duration 0.1 sec, intensity 200 mW). Rupture of Bruch's membrane was confirmed by the generation of air bubbles during laser application. In this specification, the number n in the figures showing the results of the laser CNV test indicates the number of sites evaluated for laser irradiation. The mice were irradiated with laser on the 42nd and 56th days after the start of feeding, respectively. On the 7th day after laser irradiation, the mice were anesthetized with Somnopentyl (Kyoritsu Seiyaku Co., Ltd., Tokyo, Japan), and the eyeballs were enucleated. Only the retinal pigment epithelium was flat-mounted and fixed with 4% PFA (prepared by mixing 4% paraformaldehyde, 16% PFA (ELECTRONMICROSCOPY SCIENCE), 20x PBS, and MQ at a ratio of 5:1:14) for 1 hour, washed three times with PBS, and stained with isolectin (IsolectinGS-IB4FromGriffonia simplicifolia, Alexa FluorTM 594 Conjugate: diluted in PBS = 1:200) for 2 nights at 4°C. The mice were then washed three times with PBS and mounted with Fluoromount-G (SouthernBiotech). The mice were observed under a ZweissLSM 710 confocal microscope, and the volume of CNV was quantified with Imaris (Bitplane).

[0138] Figure 5(A) is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation on the 42nd day (6 weeks) after the start of administration. Figure 5(B) is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation on the 56th day (8 weeks) after the start of administration. As shown in Figures 5(A) and (B), even when choroidal neovascularization (CNV) was induced by laser irradiation, the Garcinia administration group showed an inhibitory effect on the induction of CNV. As predicted in Test Examples 1 and 2, it was suggested that if a component that has been confirmed to have HIF inhibitory activity in retinal cells has an inhibitory effect on the induction of CNV when choroidal neovascularization (CNV) is induced by laser irradiation, it will have an inhibitory effect on the induction of CNV.

[0139] [Test Example 5: Laser CNV Test Method (Angiogenesis Inhibition) 2] Six-week-old C57BL6 / J mice (male) were intraperitoneally administered corn oil (control group) and (-)-hydroxycitric acid suspended in corn oil (30 mg / kg / day / 1000 μl) (5 days / week) for a total of 2 weeks. A choroidal neovascularization (CNV) model was created by irradiating the mouse with a 200 mW, 0.1 sec, 75 μm laser in the first week of administration, and the CNV volume was quantified 7 days after irradiation.

[0140] Specifically, C57BL / 6 mice were anesthetized with MMB, and an argon laser (spot size 75 μm, duration 0.1 sec, intensity 200 mW) was applied to five locations near the optic disc per eye. Rupture of Bruch's membrane was confirmed by the generation of air bubbles during laser application. Laser irradiation was performed one week after the start of administration. On the seventh day after laser irradiation, the mice were anesthetized with Somnopentyl (Kyoritsu Seiyaku Co., Ltd., Tokyo, Japan), and the eyeballs were enucleated. Only the retinal pigment epithelium was flat-mounted and fixed in 4% PFA (prepared by mixing 4% paraformaldehyde, 16% PFA (ELECTRONMICROSCOPY SCIENCE), 20x PBS, and MQ in a ratio of 5:1:14) for 1 hour. After washing three times with PBS, the mice were incubated with isolectin (IsolectinGS-IB4 From Griffonia simplicifolia, Alexa Fluor 9781). TMThe sections were stained with 594 conjugate:PBS (diluted 1:200) at 4°C for 2 nights. Then, the sections were washed three times with PBS and mounted with Fluoromount-G (SouthernBiotech). The sections were observed under a Zweiss LSM 710 confocal microscope, and the CNV volume was quantified using Imaris (Bitplane).

[0141] FIG. 6 is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation one week after the start of administration. As shown in Figure 6, even when choroidal neovascularization (CNV) was induced by laser irradiation, the hydroxycitric acid administration group showed an inhibitory effect on CNV induction. Considering the results of Test Examples 1, 2, and 5, it was suggested that a component that has been confirmed to have HIF inhibitory activity in retinal cells would have an inhibitory effect on CNV induction when choroidal neovascularization (CNV) was induced by laser irradiation.

[0142] [Test Example 6: Laser CNV Test Method (Inhibition of Angiogenesis) 3] The diet was orally administered to 4-week-old C57BL6 / J mice (male) for a total of 63 days, and on the 56th day after the start of administration, a choroidal neovascularization (CNV) model was created by irradiating the mouse with a laser at 200 mW, 0.1 sec, and 75 μm, and the CNV volume was quantified 7 days after irradiation. The control group was administered a control diet (MF, Oriental Yeast Co., Ltd.), and the carrot extract group was administered a diet containing carrot extract (Matsuura Pharmaceutical Co., Ltd.) mixed with the control diet at 200 mg / kg / day.

[0143] Specifically, C57BL / 6 mice were anesthetized with MMB, and 5 sites per eye were irradiated near the optic disc with an argon laser (spot size 75 μm, duration 0.1 sec, intensity 200 mW). Rupture of Bruch's membrane was confirmed by the generation of air bubbles during laser application. In this specification, the number n in the figures showing the results of the laser CNV test indicates the number of sites evaluated for laser irradiation. The mouse was irradiated with a laser on the 56th day after the start of feeding, and on the 7th day after laser irradiation, after anesthetizing with Somnopentyl (Kyoritsu Seiyaku Co., Ltd., Tokyo, Japan), the eyeball was enucleated, and only the retinal pigment epithelium was flat-mounted and fixed with 4% PFA (prepared by mixing 4% paraformaldehyde, 16% PFA (ELECTRONMICROSCOPYSCIENCE), 20x PBS, and MQ at a ratio of 5:1:14) for 1 hour, washed three times with PBS, and stained with isolectin (IsolectinGS-IB4From Griffonia simplicifolia, Alexa FluorTM 594 Conjugate: diluted with PBS = 1:200) for 2 nights at 4°C. After that, it was washed three times with PBS and mounted with Fluoromount-G (SouthernBiotech). This was observed under a ZweissLSM 710 confocal microscope, and the volume of CNV was quantified with Imaris (Bitplane).

[0144] FIG. 7 is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation 8 weeks after the start of administration. As shown in Figure 7, even when choroidal neovascularization (CNV) was induced by laser irradiation, the carrot extract administration group showed an inhibitory effect on CNV induction. Considering the results of Test Examples 1, 2, and 6, it was suggested that if a component has been confirmed to have HIF inhibitory activity in retinal cells, it would have an inhibitory effect on CNV induction when choroidal neovascularization (CNV) is induced by laser irradiation.

[0145] [Test Example 7: Laser CNV Test Method (Inhibition of Angiogenesis) 4] Three-week-old C57BL6 / J mice (male) were administered PBS (control group) and lactoferrin suspended in PBS (1600mg / kg / day) (lactoferrin administration group) 6 days / week for a total of 5 weeks via gastric tube. A choroidal neovascularization (CNV) model was created by irradiating the mouse with a 200mW, 0.1sec, 75μm laser one week after administration, and the CNV volume was quantified one week after irradiation.

[0146] Specifically, C57BL / 6 mice were anesthetized with MMB, and an argon laser (spot size 75 μm, duration 0.1 sec, intensity 200 mW) was applied to five locations near the optic disc per eye. Rupture of Bruch's membrane was confirmed by the generation of air bubbles during laser application. Four weeks after the start of administration, the mouse was irradiated with a laser. One week after laser irradiation, the mouse was anesthetized with Somnopentyl (Kyoritsu Seiyaku Co., Ltd., Tokyo, Japan), and the eyeball was removed. Only the retinal pigment epithelium was flat-mounted and fixed in 4% PFA (prepared by mixing 4% paraformaldehyde, 16% PFA (ELECTRON MICROSCOPY SCIENCE), 20x PBS, and MQ in a ratio of 5:1:14) for 1 hour. After washing three times with PBS, the mouse was incubated with isolectin (IsolectinGS-IB4From Griffonia simplicifolia, Alexa Fluor 9781). TM The sections were stained with 594 Conjugate:PBS (diluted 1:200) for 2 nights at 4°C. Then, the sections were washed three times with PBS and mounted with Fluoromount-G (SouthernBiotech). The sections were observed under a Zweiss LSM 710 confocal microscope, and the CNV volume was quantified using Imaris (Bitplane).

[0147] FIG. 8 is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation 4 weeks after the start of administration. As shown in Figure 8, even when choroidal neovascularization (CNV) was induced by laser irradiation, the lactoferrin administration group showed an inhibitory effect on CNV induction. Considering the results of Test Examples 1, 2, and 7, it was suggested that if a component that has been confirmed to have HIF inhibitory activity in retinal cells is used, it will have an inhibitory effect on CNV induction when choroidal neovascularization (CNV) is induced by laser irradiation.

[0148] [Test Example 8: Laser CNV Test Method (Inhibition of Angiogenesis) 5] Three-week-old C57BL6 / J mice (male) were orally administered food for a total of 63 days. On the 56th day after the start of administration, a choroidal neovascularization (CNV) model was created by irradiating the mouse with a laser at 200 mW, 0.1 sec, and 75 μm. The CNV volume was quantified 7 days after irradiation. The control group was administered vitamin B 6 Vitamin B deficiency AIN-93G (Oriental Yeast Co., Ltd.) 6 The diet was mixed with vitamin B at 0.17 mg / kg / day. 6 The administration group was vitamin B 6 Deficiency of AIN-93G and Vitamin B 6 The rice bran group was administered 1.53 mg / kg / day or 5.96 mg / kg / day of vitamin B 6 Deficiency of AIN-93G and Vitamin B 6 The rats were fed a diet containing 0.17 mg / kg / day of glycerol and 100 mg / kg / day of defatted rice bran (Oryza Oil & Fat Chemical Co., Ltd.).

[0149] Specifically, C57BL / 6 mice were anesthetized with MMB, and 5 sites per eye were irradiated near the optic disc with an argon laser (spot size 75 μm, duration 0.1 sec, intensity 200 mW). Rupture of Bruch's membrane was confirmed by the generation of air bubbles during laser application. In this specification, the number n in the figures showing the results of the laser CNV test indicates the number of sites evaluated for laser irradiation. The mice were irradiated with a laser on the 56th day after the start of feeding, and on the 7th day after laser irradiation, the mice were anesthetized with Somnopentyl (Kyoritsu Seiyaku Co., Ltd., Tokyo, Japan), and the eyeballs were enucleated. Only the retinal pigment epithelium was flat-mounted and fixed with 4% PFA (prepared by mixing 4% paraformaldehyde, 16% PFA (ELECTRONMICROSCOPY SCIENCE), 20x PBS, and MQ at a ratio of 5:1:14) for 1 hour, washed three times with PBS, and stained with isolectin (IsolectinGS-IB4From Griffonia simplicifolia, Alexa FluorTM 594 Conjugate: diluted in PBS = 1:200) for 2 nights at 4°C. The mice were then washed three times with PBS and mounted with Fluoromount-G (SouthernBiotech). The mice were observed under a ZweissLSM 710 confocal microscope, and the volume of CNV was quantified with Imaris (Bitplane).

[0150] FIG. 9 is a graph showing the results of choroidal neovascularization (CNV) induction after laser irradiation 8 weeks after the start of administration. As shown in Figure 9, even when choroidal neovascularization (CNV) was induced by laser irradiation, vitamin B 6 In the group administered with nicotinamide and the group administered with rice bran, the effect of suppressing the induction of CNV was observed. Considering the results of Test Examples 1, 2, and 8, it was suggested that if a component that has been confirmed to have HIF inhibitory activity in retinal cells is used, it will have the effect of suppressing the induction of CNV when choroidal neovascularization (CNV) is induced by laser irradiation.

[0151] [Test Example 9: Test to confirm expression of HIF downstream genes] Human retinal pigment epithelial cell line (ARPE) was added to a 12-well plate at 1.0 × 10 6 After 24 hours, the cells were attached to the bottom of the plate, and the medium was replaced with 200 μM CoCl. 2 At the same time, Garcinia cambogia extract (Bioactives Japan Co., Ltd.) and (-)-hydroxycitric acid calcium salt were added at 1 mg / ml.

[0152] Eight hours after addition, RNA was extracted and gene expression was confirmed by RT-PCR (ReverTra AceqPCR RT Master Mix (TOYOBO) and real-time PCR (THUNDERBIRDSYBO qPCR Mix (TOYOBO)). The results for Garcinia cambogia extract (Bioactives Japan Co., Ltd.) and (-)-hydroxycitric acid calcium salt are shown in Figure 10 and Figure 11, respectively.

[0153] As shown in Figures 10 and 11, CoCl 2 The expression of HIF downstream genes, VEGF, GLUT1, PDK1, and BNIP3, which were upregulated in the rats, was significantly suppressed by the addition of Garcinia cambogia extract and (-)-hydroxycitric acid calcium salt.

[0154] In this specification, the MMB anesthetic was prepared by mixing three drugs: midazolam (4 mg / kg, SANDOZ, Yamagata, Japan), medetomidine (0.75 mg / kg, Fukushima, Japan), and butorphanol tartrate (5 mg / kg, Meiji Seika Pharmaceutical Co., Tokyo, Japan). Each drug was contained in the MMB anesthetic in a ratio of 1:1.07:1.33.

[0155] All procedures herein have been approved by the Ethics Committee on Animal Research at Keio University School of Medicine, which adheres to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research, the Keio University Institutional Guidelines for Animal Experimentation, and the Guidelines for the Use of Animals in Research (ARRIVE). C57BL / 6J and BALB / cA mice were obtained from CLEA Japan, Inc.

Claims

1. A composition for suppressing HIF, comprising pyridoxine and / or a salt thereof.

2. A composition for retinal protection comprising pyridoxine and / or a salt thereof.

3. A composition for suppressing high levels of HIF in retinal pigment epithelial cells, comprising pyridoxine and / or a salt thereof.

4. A composition for inhibiting VEGF in the eye, comprising pyridoxine and / or a salt thereof.

5. A composition for inhibiting ocular neovascularization, comprising pyridoxine and / or a salt thereof.

6. A composition for maintaining homeostasis of vascular tissue by inhibiting VEGF in the eye and inhibiting angiogenesis induced by VEGF, comprising pyridoxine and / or a salt thereof.

7. A composition for reducing the risk of, preventing, or treating a posterior ocular disease, comprising pyridoxine and / or a salt thereof.

8. The composition according to claim 7, wherein the posterior ocular disease is at least one selected from the group consisting of age-related macular degeneration, diabetic retinopathy, diabetic macular edema, retinitis pigmentosa, proliferative vitreous retinopathy, retinal artery occlusion, retinal vein occlusion, uveitis, Leber's disease, retinopathy of prematurity, retinal detachment, retinal pigment epithelial detachment, central serous chorioretinopathy, central exudative chorioretinopathy, polypoidal choroidal vasculopathy, multifocal choroiditis, neovascular maculopathy, retinal aneurysm, retinal angiomatous proliferation, optic neuropathy caused by these diseases, optic neuropathy caused by glaucoma, and ischemic optic neuropathy.

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