JP7201191B1 - Erythropoietin production accelerator - Google Patents

Abstract

A novel erythropoietin production promoter is provided. [Means for Solving Problem] An erythropoietin production promoter characterized by containing a processed product of seabuckthorn fruit as an active ingredient. [Selection diagram] Fig. 1

Description

TECHNICAL FIELD The present invention relates to an erythropoietin production promoter.

Erythropoietin (EPO) is a glycoprotein hormone that stimulates differentiation induction of erythroid stem cells (progenitor cells) and promotes red blood cell production. Its production takes place mainly in the kidney and is regulated according to the partial pressure of oxygen in arterial blood. Administration of erythropoietin is expected to treat renal anemia, improve fatigue, improve exercise endurance, etc., and erythropoietin preparations such as recombinant artificial erythropoietin and erythropoietin-like compounds are known. A compound that promotes the production of erythropoietin in the body has also been proposed (Patent Document 1).

Japanese Patent Application Laid-Open No. 2020-132561

An object of the present invention is to provide a novel erythropoietin production promoter.

As a result of earnest research to solve the above problems, the inventors of the present invention have found that the following inventions meet the above objects, and have completed the present invention.

That is, the present invention relates to the following inventions.
<1> An erythropoietin production promoter characterized by containing a processed product of seabuckthorn fruit as an active ingredient.
<2> The erythropoietin production promoter according to <1> above, wherein the processed sea buckthorn fruit is sea buckthorn juice and/or dried sea buckthorn juice.
<3> The erythropoietin production promoter according to <1> or <2> above, wherein the sea buckthorn fruit is a sea buckthorn fruit containing pericarp, pulp and seeds.
<4> The erythropoietin production promoter according to any one of <1> to <3>, which is an oral composition.

According to the present invention, a novel erythropoietin production promoter is provided.

FIG. 1 shows erythropoietin concentrations of Example 1 and a control study. In the figure, * indicates that p<0.05.

Embodiments of the present invention will be described in detail below. is not limited to the contents of In this specification, when the expression "~" is used, it is used as an expression including the numerical values before and after it.

<Erythropoietin production promoter of the present invention>
TECHNICAL FIELD The present invention relates to an erythropoietin production-enhancing agent containing a processed seabuckthorn fruit as an active ingredient. The present inventors have newly found that a processed product of seabuckthorn fruit can promote the production of erythropoietin. The present invention is based on this finding.

(processed sea buckthorn fruit)
Sea buckthorn (scientific name: Hippophae rhamnoides L., English name: Sea buckthorn) is a plant of the genus Hippophae of the family Gumaceae native to the Eurasian Continent, and is said to have grown more than 70 million years ago. The processed sea buckthorn fruit is obtained by processing the sea buckthorn fruit. The fruit may be used in a state containing the pericarp and seeds, or may be used with the pericarp removed, or only the pulp portion from which the pericarp and seeds have been removed may be used. Examples of the processed sea buckthorn fruit include sea buckthorn juice, dried sea buckthorn juice, dried sea buckthorn fruit, and sea buckthorn fruit extract. Sea buckthorn juice and/or dried sea buckthorn juice are preferred.

(sage juice)
Sea buckthorn juice is fruit juice obtained by subjecting sea buckthorn fruit to pressure (pressing), concentrated juice obtained by concentrating this juice, or diluted juice obtained by diluting this juice. Sea buckthorn juice may be liquid, semi-liquid such as puree or paste, or a mixture thereof. Processing to apply pressure (compressing processing) includes crushing processing and grinding processing. You can get juice.

The sea buckthorn juice may be used after subjecting the sea buckthorn fruit to pressure treatment, and then centrifuging, filtering, or straining as appropriate. Straining can be performed, such as by passing through a mesh (eg, 40-60 mesh). Puree removes at least a portion of the fibrous material of the pericarp, seeds, and pulp.

The sea buckthorn juice is preferably a juice obtained from a sea buckthorn fruit containing a pericarp, more preferably a fruit juice obtained from a fruit containing a pericarp, pulp and seeds. For example, a liquid or semi-liquid product obtained by processing sea buckthorn fruit containing skin, pulp and seeds under pressure, sea buckthorn fruit containing skin, pulp and seeds under pressure, and further Liquids or semi-liquids obtained by pureeing, or mixtures thereof, can be used as sea buckthorn juice.

(Dried sea bass juice)
The dried sea sage juice is obtained by drying sea sage juice. Drying can be performed by a known drying method such as sun drying, heat drying, reduced pressure drying, freeze drying, spray drying, and the like. Moreover, the dried product is preferably pulverized or pulverized during drying using spray drying or the like to obtain a dry powder.

(subject)
Subjects for use of the erythropoietin production-enhancing agent of the present invention are animals including humans. Animals other than humans include dogs and cats.

Since the processed sage fruit has an erythropoietin production-enhancing effect, the erythropoietin production-enhancing agent of the present invention can be used for the prevention and treatment of anemia such as renal anemia. Renal anemia is different from iron deficiency anemia caused by inadequate hemoglobin production due to iron deficiency. anemia caused by Specifically, the erythropoietin production promoter of the present invention may be used by subjects already suffering from renal anemia or anemia to improve renal anemia or anemia. It can also be used for the prevention of renal anemia or anemia in a subject.

The processed sage fruit can be used as it is in a product, or can be blended in a product. So far, few compounds have been known that can promote the production of erythropoietin by simply ingesting them. Processed sage fruit is derived from natural products, so it is safe and highly palatable, and can be easily processed into a form that is easily ingested on a daily basis. Therefore, the erythropoietin production-enhancing agent of the present invention is preferably an oral composition. The processed product of sea buckthorn fruit can be used as a product as an oral composition, or the processed sea buckthorn fruit can be blended with the composition for oral use to obtain an oral composition containing the processed sea buckthorn fruit. . The use as an oral composition is not particularly limited, and it can be used as a food composition, a pharmaceutical composition for oral administration, and the like.

(Food composition)
By making the processed sage fruit into a food composition that is easily orally ingested on a daily basis, it is also easy to ingest for a long period of time. Therefore, it is preferable to prepare a food composition for promoting erythropoietin production containing a processed product of sea buckthorn fruit as an active ingredient. This food composition can be used not only for general foods but also for the purpose of maintaining and promoting health, such as foods for special dietary uses, foods for specified health uses, foods with function claims, foods with nutrient function claims, nutritional supplements, health supplements, and supplements. It also includes food and/or beverages consumed. Among these, food with specified health uses, food with functional claims, and food with nutrient function claims, which are foods with health claims, are preferred.

The shape of the food composition is not particularly limited, and may be any shape such as solid, liquid, semi-solid, gel, and sol. Sea buckthorn juice can be used as it is and its content can be increased, so it is preferably in the form of a drink (liquid drink, puree, etc.).

When it is commercialized as a food composition, it may be a product made of processed sea cucumber fruit, or it may be blended in foods and beverages. In the case of commercialization as a food composition, various additives used in foods and beverages may be added in addition to the processed sea buckthorn fruit. Specifically, coloring agents, preservatives, thickeners, antioxidants, bleaching agents, antibacterial and antifungal agents, acidulants, seasonings, emulsifiers, reinforcing agents, manufacturing agents, flavors, sweeteners, etc. may be added.

Foods and beverages that are objects of the food composition are not particularly limited. Examples of foods include foods such as sausages, hams, processed seafood products, jellies, candies, and chewing gums. Examples of beverages include various fruit juice beverages, teas, soft drinks, alcoholic beverages, energy drinks, and the like.

It may also be used as an animal food composition, and the processed sage fruit can also be added to an animal food composition such as pet food.

(Pharmaceutical composition)
The processed sea buckthorn fruit can be mixed with a pharmaceutically acceptable carrier to prepare a pharmaceutical composition for oral administration. In addition, in the present invention, the pharmaceutical composition includes not only pharmaceuticals but also quasi-drugs. The dosage form of the pharmaceutical composition is not particularly limited, and may be a solid formulation or a liquid formulation. Specific examples include powders, granules, tablets, capsules, troches, suspensions, emulsions, syrups, elixirs and the like. Pharmaceutically acceptable carriers include, for example, binders, stabilizers, excipients, diluents, pH buffers, thickeners, colorants, dispersants, emulsifiers, suspending agents, preservatives, Disintegrants, solubilizers, solubilizers and the like are included.

The amount of the processed sea buckthorn fruit blended in the composition (food composition, pharmaceutical composition, etc.) containing the processed sea buckthorn fruit varies depending on the mode and the like, and is appropriately adjusted. For example, the content of the processed sea buckthorn fruit in the composition containing the processed sea buckthorn fruit (in terms of solid content) is 1% by mass or more, 2% by mass or more, 3% by mass or more, 4% by mass or more, 5 mass % or more is preferable. The upper limit is not particularly limited, but the upper limit may be appropriately set such that the content of the processed seabuckthorn fruit (in terms of solid content) is 20% by mass or less, or 15% by mass. The solid content of the processed seabuckthorn fruit is the mass obtained by removing water and solvent from the processed seabuckthorn fruit. For example, sea buckthorn juice contains about 80 to 90% by mass of water, so the amount of solids is about 10 to 20% by mass. The content of the processed sea buckthorn fruit (in terms of solid content) is the percentage obtained by dividing the solid content of the processed sea buckthorn fruit by the mass of the composition containing the processed sea buckthorn fruit.

In addition, when the sea buckthorn juice is used as a processed sea buckthorn fruit, the sea buckthorn juice can be used as it is, or a composition containing sea buckthorn juice mixed with additives and the like can be used as an oral composition for promoting erythropoietin production. . The content of sea buckthorn juice in the composition for oral use (mass of sea buckthorn juice/mass of oral composition×100(%)) is 30% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, The higher the content, in the order of 80% by mass or more and 90% by mass or more, the more preferable. Moreover, the upper limit of the content of sea buckthorn juice in the composition for oral use may be appropriately set to 100% by mass or less, 99.99% by mass or less, or 99.98% by mass.

The intake amount of the composition (food composition, pharmaceutical composition, etc.) containing the processed sage fruit may be appropriately selected according to the subject's sex, body weight, age, purpose of use, and the like. For example, in the case of an adult human, the daily intake of processed seabuckthorn fruit (converted to dry solid content) can be 0.1 to 45 g, 1 to 35 g, 2 to 15 g, or the like. In the case of a composition containing sea buckthorn juice, for example, the daily intake for an adult human can be 1 to 300 g, 10 to 250 g, 15 to 200 g, 20 to 100 g, and the like. In addition, the ingestion can be carried out once or several times a day, and the ingestion may be continued for several days to several weeks or several months.

EXAMPLES 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 unless the gist thereof is changed.

[Example]
I. Preparation of sea buckthorn juiceH. rhamnoides fruits (flesh and skin, from Banhan Mountain, Inner Mongolia) were frozen at −18° C. with branches and leaves, removed branches and leaves in the frozen state, and washed with drinking water. Removed damaged fruit. Next, the fruit with the peel attached was put into a mixer, pureed, filtered (60 mesh) to remove the peel residue and seeds, and subjected to high-temperature flash sterilization to obtain sea buckthorn juice. Stevia was added to the sea buckthorn juice, mixed while removing foreign matter with a mesh (40 mesh), and heat sterilized to obtain sea buckthorn juice containing 99.95% by mass of sea buckthorn juice.

II. Measurement of EPO Concentration The freeze-dried sea buckthorn juice obtained in I above was added to Hep-G2 cells, and the amount of erythropoietin (EPO) produced in the cell supernatant was evaluated.

(i) Preparation of Sample Solution Pre-frozen sea buckthorn juice was placed in a freeze dryer and allowed to stand under conditions of −50° C. and 40 Pa for several days to obtain a freeze-dried product. Lyophilized sea buckthorn juice was dissolved in sterile water to 0.02 mg/mL, 0.2 mg/mL, 2 mg/mL, 20 mg/mL, 40 mg/mL and 80 mg/mL. Then, before adding the sample to human liver cancer-derived cell line (HepG2 cells), it was diluted 200-fold with serum-free EMEM (containing 1% penicillin-streptomycin) (final concentrations 0.1 μg/mL, 1 μg/mL, 10 μg/mL). mL, 100 μg/mL, 200 μg/mL, and 400 μg/mL). 100 μL of the 200-fold diluted sample solution was added to the cells.

(ii) Test Human hepatoma-derived cell line (HepG2 cells) was grown to φ10 cm until confluent using Eagle's minimal essential Medium (EMEM) (containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS)). It was pre-cultured in a dish. Thereafter, the cells were washed with phosphate buffered saline (PBS), resuspended in a medium, seeded in a 96-well plate at a concentration of 0.5×10 ⁵ cells/well, and placed in a CO ₂ incubator (37° C., 5% CO). ₂ ) and cultured overnight.
After addition of the sample solution prepared in (i) above, the cells were again cultured in the CO ₂ incubator (37° C., 5% CO ₂ ) for 24 hours.

(iii) Control test Sterilized water was used instead of the sample solution, and the test was carried out in the same manner as in (ii) above.

(iv) Evaluation of Erythropoietin Production Amount by ELISA Method HepG2 cell culture supernatant was collected 24 hours after the addition of the evaluation sample. An Enzyme-Linked Immunosorbent Assay (ELISA) kit for Erythropoietin (EPO) (Cloud-Clone Crop.) was used to calculate the amount of EPO produced in the culture supernatant. EPO standards, samples and blanks (buffer only without sample) were added to the antibody coated plates. After that, each sample was removed, detection reagent A was added, and the mixture was allowed to stand at 37° C. for 1 hour. Detection Reagent A was removed and the wells were washed three times with wash reagent. Subsequently, detection reagent B was added, and the mixture was allowed to stand at 37° C. for 30 minutes. Detection Reagent B was removed and the wells were washed 5 times with wash reagent. A substrate solution was added, and the mixture was allowed to stand at 37° C. for 20 minutes in the dark. A reaction stopping solution was added, and the absorbance at 450 nm was measured with a microplate reader.

Figure 1 shows the erythropoietin concentrations of Example 1 and the control study. As shown in FIG. 1, the addition of freeze-dried seaweed juice (200 μg/mL treatment group) significantly promoted EPO production compared to the control (p<0.05). An increase in EPO production was also observed in the 1 μg/mL treated area, the 10 μg/mL treated area, the 100 μg/mL treated area and the 400 μg/mL treated area as compared to the control.

Since sea buckthorn juice contains the same components as the freeze-dried sea buckthorn juice except water, similar effects can be expected with sea buckthorn juice. The water content of sea buckthorn juice obtained in I above was 86% by mass.

INDUSTRIAL APPLICABILITY The erythropoietin production-enhancing agent of the present invention can be used in foods, pharmaceuticals, and the like, and is industrially useful.

Claims (2)

An erythropoietin production-enhancing agent characterized by containing sea buckthorn juice and/or dried sea buckthorn juice as an active ingredient. The erythropoietin production promoter according to claim 1 , which is an oral composition.

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