Erythropoietin production promoter

Abstract

To provide a novel erythropoietin production promoter.SOLUTION: An erythropoietin production promoter contains isorhamnetin and / or an isorhamnetin glycoside as an active ingredient.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to an erythropoietin production promoter. [Background technology]

[0002] Erythropoietin (EPO) is a glycoprotein hormone that stimulates differentiation of erythroid stem cells (progenitor cells) and promotes red blood cell production. Its production is primarily in the kidneys and is regulated according to the oxygen partial pressure in arterial blood. Administration of erythropoietin is expected to treat renal anemia, alleviate fatigue, and improve exercise endurance, and erythropoietin preparations such as recombinant artificial erythropoietin and erythropoietin-like compounds are known. Compounds and bacteria that promote erythropoietin production in the body have also been proposed (e.g., Patent Document 1). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Japanese Patent Publication No. 2020-132561 Summary of the Invention [Problem to be solved by the invention]

[0004] An object of the present invention is to provide a novel erythropoietin production promoter. [Means for solving the problem]

[0005] The present inventors have conducted extensive research to solve the above problems and have found that the following inventions meet the above objectives, thereby completing the present invention. <1> An erythropoietin production promoter containing isorhamnetin and / or isorhamnetin glycoside as an active ingredient. <2> The composition is an oral composition. <1> The erythropoietin production promoter according to claim 1. <3> The composition contains isorhamnetin and / or isorhamnetin glycoside so that the total amount of the isorhamnetin and the amount of the isorhamnetin glycoside converted into isorhamnetin aglycone is 0.01 μg to 10,000 μg as the daily intake amount for an adult human. <1> or <2> The erythropoietin production promoter according to claim 1. [Effects of the Invention]

[0006] According to the present invention, a novel erythropoietin production promoter is provided. [Brief explanation of the drawings]

[0007] [Figure 1] FIG. 1 shows the amount of EPO produced (%) per cell by the addition of isorhamnetin. DETAILED DESCRIPTION OF THE INVENTION

[0008] The following describes in detail the embodiments of the present invention, but the explanation of the constituent elements described below is one example (typical example) of the embodiment of the present invention, and the present invention is not limited to the following content unless the gist of the present invention is changed. Note that when the expression "to" is used in this specification, it is used as an expression including the numerical values ​​or physical property values ​​before and after it.

[0009] <Erythropoietin production promoter> The present invention relates to an erythropoietin production enhancer containing isorhamnetin and / or isorhamnetin glycoside as an active ingredient (hereinafter, sometimes referred to as "the EPO production enhancer of the present invention").

[0010] Isorhamnetin is a compound classified as an O-methylated flavonol. Isorhamnetin glycosides are compounds in which sugars such as glucose and rhamnose are bound to isorhamnetin and are converted to isorhamnetin aglycone (isorhamnetin) by metabolism. Specific examples of isorhamnetin glycosides include narcisine (isorhamnetin-3-O-rutinoside), isorhamnetin-3-O-glucoside-7-O-rhamnoside, isorhamnetin-3-O-glucoside, isorhamnetin-3-O-rutinoside-7-O-glucoside, and isorhamnetin-3-O-rutinoside-4'-O-glucoside. One or more isorhamnetin glycosides may be combined.

[0011] When the EPO production promoter of the present invention contains isorhamnetin, the contained isorhamnetin can promote erythropoietin production. Furthermore, when the EPO production promoter of the present invention contains isorhamnetin glycoside, the contained isorhamnetin glycoside is metabolized in vivo to produce isorhamnetin aglycone (isorhamnetin), and the produced isorhamnetin aglycone can promote erythropoietin production. Isorhamnetin aglycone is obtained by removing sugars from isorhamnetin glycoside and has the same structure as isorhamnetin. Therefore, the EPO production promoter of the present invention is used to promote erythropoietin production by isorhamnetin and / or isorhamnetin aglycone (isorhamnetin) produced from isorhamnetin glycoside.

[0012] (Oral Composition) The EPO production-promoting agent of the present invention can be used as a food composition or a pharmaceutical composition, and is preferably in the form of an oral composition. That is, the EPO production-promoting agent of the present invention is preferably in the form of a food composition containing isorhamnetin and / or isorhamnetin glycoside as an active ingredient for promoting erythropoietin production, or an oral pharmaceutical composition containing isorhamnetin and / or isorhamnetin glycoside as an active ingredient for promoting erythropoietin production.

[0013] (Food composition) The EPO production-promoting agent of the present invention is preferably formulated as a food composition. Formation of the food composition facilitates long-term daily intake. When formulated as a food composition, examples of the food composition include a composition in which isorhamnetin and / or isorhamnetin glycoside itself is incorporated into the food composition, a processed product (e.g., pulverized material, crushed material, juice, puree, extract, dried material, etc.) of a plant (whole or organ) containing isorhamnetin and / or isorhamnetin glycoside, and a composition in which these processed products are incorporated into the food composition. Such food compositions include not only general foods but also foods and / or beverages taken for the purpose of maintaining or improving health, such as foods for specified health uses, foods with functional claims, foods with nutrient functions, dietary supplements, health supplements, and supplements. Among these, foods with health claims, such as foods for specified health uses, foods with functional claims, and foods with nutrient functions, are preferred.

[0014] The form of the food composition is not particularly limited, and may be any of solid, liquid, semi-solid, gel, sol, etc. Foods and beverages are not particularly limited, and examples of foods include sausage, ham, processed seafood products, jelly, candy, chewing gum, etc. Beverages include various fruit juice drinks, teas, soft drinks, alcoholic beverages, energy drinks, etc. Food compositions may also contain additives such as colorants, preservatives, thickening stabilizers, antioxidants, bleaching agents, antibacterial and antifungal agents, acidulants, seasonings, emulsifiers, strengthening agents, flavorings, and sweeteners.

[0015] (Pharmaceutical composition) The EPO production enhancer of the present invention can be made into a pharmaceutical composition. This pharmaceutical composition includes both drugs and quasi-drugs. When made into a pharmaceutical composition, the EPO production enhancer of the present invention basically contains isorhamnetin and / or isorhamnetin glycoside or a material containing them, and a pharmaceutically acceptable carrier. Examples of such a pharmaceutical composition include a pharmaceutical composition in which isorhamnetin and / or isorhamnetin glycoside itself is incorporated, a processed product (e.g., pulverized product, crushed product, juice, puree, extract, dried product, etc.) of a plant (whole or organ) containing isorhamnetin and / or isorhamnetin glycoside, and a pharmaceutical composition in which such a processed product is incorporated.

[0016] The dosage form of the pharmaceutical composition is not particularly limited and may be a solid or liquid preparation. Specific examples include powders, granules, tablets, capsules, lozenges, suspensions, emulsions, syrups, and elixirs. Pharmaceutically acceptable carriers include binders, stabilizers, excipients, diluents, pH buffers, thickeners, colorants, dispersants, emulsifiers, suspending agents, preservatives, disintegrants, solubilizers, and dissolution aids.

[0017] The content of isorhamnetin and / or isorhamnetin glycoside in the EPO production promoter of the present invention is designed according to the method of ingestion, etc., so that the sum of the amount of isorhamnetin and the amount of isorhamnetin glycoside converted into isorhamnetin aglycone is the desired daily intake for an adult human. For example, the EPO production promoter of the present invention preferably contains isorhamnetin and / or isorhamnetin glycoside so that the total amount of isorhamnetin and the amount of isorhamnetin glycoside converted into isorhamnetin aglycone is 0.01 μg to 10,000 μg, 0.1 μg to 9,500 μg, 1 μg to 9,000 μg, 10 μg to 8,000 μg, 100 μg to 7,000 μg, 1,000 μg to 6,000 μg, or 1,500 μg to 5,000 μg, as the daily intake for an adult human. When the EPO production promoter of the present invention contains only isorhamnetin, the "total amount of isorhamnetin and the amount of isorhamnetin glycoside converted into isorhamnetin aglycone" is the same as the amount of isorhamnetin, and when the EPO production promoter of the present invention contains only isorhamnetin glycoside, the "total amount of isorhamnetin and the amount of isorhamnetin glycoside converted into isorhamnetin aglycone" is the same as the amount of isorhamnetin glycoside converted into isorhamnetin aglycone. As mentioned above, isorhamnetin aglycone is a form obtained by removing sugars from isorhamnetin glycoside, so the amount of isorhamnetin glycoside converted into isorhamnetin aglycone can be calculated from the amount of isorhamnetin glycoside and its structure. When two or more isorhamnetin glycosides are contained, the amount of isorhamnetin aglycone is the sum of the amounts of each isorhamnetin glycoside converted into isorhamnetin aglycone.

[0018] The EPO production enhancer of the present invention may be taken once a day or in divided doses (e.g., 2 to 4 times) per day to achieve the above-mentioned intake amount. The daily intake amount refers to the total amount ingested or administered in one day, and when taken in divided doses per day, it is the total amount of these doses. Furthermore, the EPO production enhancer of the present invention can be taken continuously, for example, over several days to several weeks or months.

[0019] The EPO production promoters of the present invention are not limited to humans, but may also be used in non-human animals such as dogs and cats. Therefore, the EPO production promoters of the present invention may be incorporated into animal food compositions such as pet foods or veterinary pharmaceutical compositions. Specifically, they may be incorporated into animal food compositions or veterinary pharmaceutical compositions containing isorhamnetin or isorhamnetin glycoside itself, or into processed products (e.g., pulverized, crushed, compressed, extracts, etc.) of plants (whole or organs) containing isorhamnetin or isorhamnetin glycoside, or into animal food compositions or veterinary pharmaceutical compositions containing these processed products. [Example]

[0020] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples as long as the gist of the present invention is not changed.

[0021] Measurement of EPO concentration is a useful test for understanding the state of erythropoiesis in vivo. We evaluated whether isorhamnetin has a hematopoietic effect by adding isorhamnetin and measuring the EPO concentration in the culture supernatant.

[0022] <Experimental Method> ·Cell culture Human hepatocellular carcinoma-derived cell line (HepG2 cells) was precultured in Eagle's minimal essential medium (EMEM) (containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS)) in a 10 cm dish until confluent. After washing with phosphate-buffered saline (PBS), the cells were resuspended in medium and plated at 0.5 × 10 cells per well in a 96-well plate. 5 The cells were seeded at a density of 1000 cells / well and cultured overnight in a CO2 incubator (37°C, 5% CO2).

[0023] Adding evaluation samples Isorhamnetin (EXTRASYNTHESE, 1120 S) was dissolved in DMSO (Dimethyl Sulfoxide, Fujifilm Wako Pure Chemical Industries, Ltd., 043-07216) and added to serum-free EMEM (containing 1% penicillin-streptomycin) to prepare serum-free EMEM containing isorhamnetin at the concentrations shown in Figure 1. 24 hours after seeding of HepG2 cells, 100 μL of the prepared serum-free EMEM (containing 1% penicillin-streptomycin) containing isorhamnetin was added to each well, and the cells were cultured in a CO2 incubator (37°C, 5% CO2) for 24 hours. As a control, serum-free EMEM (containing 1% penicillin-streptomycin) to which DMSO was added was used.

[0024] -Evaluation of EPO production by ELISA method Twenty-four hours after the addition of the evaluation sample (isorhamnetin or DMSO), the HepG2 cell culture supernatant was collected. The amount of EPO produced in the HepG2 cell culture supernatant was calculated according to the protocol of the Enzyme-Linked Immunosorbent Assay (ELISA) kit for Erythropoietin (EPO) (Cloud Clone Crop.).

[0025] - Cell viability measurement by MTT method Twenty-four hours after the addition of the evaluation samples, the culture supernatant was collected, and 100 μL of sample-free EMEM (containing 1% penicillin-streptomycin) was added to each well of a 96-well plate. Next, 20 μL of MTT staining solution (5 mg / mL in PBS) was added to each well, and the plates were incubated in a CO2 incubator for 4 hours. The medium was removed, and 100 μL of hydrochloric acid-isopropanol solution was added to each well. The formazan was completely dissolved by pipetting. The absorbance at 570 nm was measured using a microplate reader, and cell viability was calculated.

[0026] Isorhamnetin was added to HepG2 cells, and the amount of erythropoietin (EPO) produced in the cell supernatant was measured and analyzed. Figure 1 shows the amount of EPO produced (%) per cell by the addition of isorhamnetin. As shown in Figure 1, the amount of EPO produced (%) per cell by the addition of isorhamnetin was increased compared to the control (DMSO). Furthermore, the addition of isorhamnetin (0.1 and 1 μg / mL treatment groups) significantly promoted EPO production compared to the control (DMSO).

Claims

1. An erythropoietin production promoter containing isorhamnetin and / or isorhamnetin glycoside as an active ingredient.

2. The erythropoietin production enhancer according to claim 1, which is an oral composition.

3. 3. The erythropoietin production promoter according to claim 1, wherein the agent contains isorhamnetin and / or isorhamnetin glycoside such that the total amount of the isorhamnetin and the amount of the isorhamnetin glycoside converted into isorhamnetin aglycone is 0.01 μg to 10,000 μg per day for an adult human.

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