JP2023182356A - Erythropoietin production accelerator - Google Patents

Abstract

To provide a novel erythropoietin production accelerator.SOLUTION: An erythropoietin production accelerator includes a treated sea buckthorn fruit product as an active ingredient.SELECTED DRAWING: Figure 1

Description

The present invention relates to an erythropoietin production promoter.

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

Japanese Patent Application Publication No. 2020-132561

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

As a result of extensive research in order to solve the above problems, the present inventors found that the following invention met the above objects, leading to 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 sardine fruit as an active ingredient.
<2> The erythropoietin production promoter according to <1> above, wherein the processed product of the seaweed fruit is a seaweed juice and/or a dried product of the seaweed fruit juice.
<3> The erythropoietin production promoter according to <1> or <2>, wherein the cane fruit is a cane 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 new erythropoietin production promoter is provided.

FIG. 2 is a diagram showing erythropoietin concentrations in Example 1 and a control test. Note that in the figure, * indicates p<0.05.

The embodiments of the present invention will be described in detail below, but the explanation of the constituent elements described below is an example (representative example) of the embodiments of the present invention, and the present invention will be described below unless the gist thereof is changed. is not limited to the content. Note that when the expression "~" is used in this specification, 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 promoter containing a processed product of Saji fruit as an active ingredient. The present inventors have newly discovered that processed products of Saji fruit can promote the production of erythropoietin. The present invention is based on this knowledge.

(Processed product of Saji fruit)
Sea buckthorn (Sea buckthorn, scientific name: Hippophae rhamnoides L., English name: Sea buckthorn) is a plant of the genus Hippophae of the Gummy family, native to the Eurasian continent, and is said to have been growing for more than 70 million years. The processed product of Japanese soybean fruit is obtained by processing the fruit of Japanese soybean. The fruit may be used in a state including the pericarp and seeds, the pericarp may be removed, or only the pulp portion from which the pericarp and seeds are removed may be used. Examples of processed products of candied fruit include candied soybean juice, dried candied fruit juice, dried candied fruit, candied fruit extract, etc., and preferably candied candied juice and/or dried candied candied fruit juice.

(Saji fruit juice)
Saji fruit juice is fruit juice obtained by applying pressure to Saji fruit (squeezing process), concentrated fruit juice obtained by concentrating this fruit juice, or diluted fruit juice obtained by diluting this fruit juice. In addition, the candied fruit juice may be a liquid, a semi-liquid such as puree or paste, or a mixture thereof. The process of applying pressure (squeezing process) includes crushing process, grinding process, etc. Specifically, it involves using one or more devices such as mixer, grinder, crusher, crusher, presser, etc. You can get fruit juice.

Saji fruit juice may be obtained by subjecting the suji fruits to pressure treatment, followed by centrifugation, filtration, or straining as appropriate. The straining can be performed by passing through a mesh (for example, 40 to 60 mesh). Pureeing removes at least some of the fibrous skin, seeds, and pulp.

It is preferable that the pericarp fruit juice is a fruit juice obtained from a pericarp containing a pericarp, and 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 pressure-processing a seaweed fruit containing pericarp, pulp and seeds; A liquid or semi-liquid product obtained by pureeing, or a mixture thereof, can be used as saji fruit juice.

(Dried sardine fruit juice)
Dried sardines juice is a product obtained by drying sardines juice. Drying can be performed by known drying methods such as solar drying, heat drying, reduced pressure drying, freeze drying, and spray drying. Further, it is preferable that the dried product is made into a dry powder by pulverizing it or pulverizing it during drying using spray drying or the like.

(subject)
The erythropoietin production promoter of the present invention is used in animals including humans. Examples of animals other than humans include dogs and cats.

Since processed products of sardine fruits have an effect of promoting erythropoietin production, the erythropoietin production promoting 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, which is caused by insufficient production of hemoglobin due to a lack of iron. Renal anemia is a condition in which the secretion of erythropoietin from the kidneys decreases due to decreased kidney function, and the ability to make red blood cells decreases. Anemia caused by. Specifically, the erythropoietin production promoter of the present invention may be used by subjects who are already suffering from renal anemia or anemia to improve renal anemia or anemia, and can be used by healthy subjects who are not yet suffering from the disease. It can also be used by subjects to prevent renal anemia and anemia.

Processed products of sardines can be made into products as they are, or can be blended into products. Until now, compounds that can promote the production of erythropoietin by easily ingesting them are not well known. Processed products of sardines are derived from natural products, so they are safe, have excellent palatability, and are easy to process into forms that are easy to consume on a daily basis. Therefore, the erythropoietin production promoter of the present invention is preferably made into an oral composition. It is possible to make an oral composition containing the processed product of the perennial fruit as it is, or by blending the processed product of the perilla fruit into the oral composition. . 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 product of sage fruit into a food composition that is easy to orally ingest on a daily basis, it is also easy to ingest over a long period of time. Therefore, it is preferable to provide a food composition for promoting erythropoietin production that contains a processed product of Saji fruit as an active ingredient. This food composition is used not only for general foods, but also for the purpose of maintaining and promoting health, such as foods for special purposes, foods for specified health uses, foods with functional claims, foods with nutrient function claims, nutritional supplements, health supplements, and supplements. Also includes ingested food and/or beverages. Among these, foods with specified health uses, foods with functional claims, and foods with nutritional function claims that 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, or sol. Since cane juice can be used as it is and its content can be increased, it is preferably in the form of a drink (liquid drink, puree, etc.).

When commercialized as a food composition, it may be a product made of processed sardine fruit, or may be incorporated into foods and drinks. In addition, in the case of commercializing the composition as a food composition, various additives used in foods and drinks may be added in addition to the processed product of sea bream fruit. Specifically, coloring agents, preservatives, thickening stabilizers, antioxidants, bleaching agents, antibacterial and antifungal agents, acidulants, seasonings, emulsifiers, fortifying agents, manufacturing agents, fragrances, sweeteners, etc. It may be added.

Foods and beverages to which the food composition is applied are not particularly limited. Examples of foods include sausages, ham, processed seafood products, jellies, candies, and chewing gum. In addition, examples of beverages include various fruit juice drinks, teas, soft drinks, alcoholic beverages, energy drinks, and the like.

Moreover, it may be used as an animal food composition, and the processed product of saji fruit can also be added to animal food compositions such as pet food.

(Pharmaceutical composition)
The processed product of sea bream fruit can be blended with a pharmaceutically acceptable carrier to form a pharmaceutical composition for oral administration. 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 preparation or a liquid preparation. Specific examples include powders, granules, tablets, capsules, troches, suspensions, emulsions, syrups, and elixirs. Pharmaceutically acceptable carriers include, for example, binders, stabilizers, excipients, diluents, pH buffers, thickeners, colorants, dispersants, emulsifiers, suspending agents, preservatives, Examples include disintegrants, solubilizers, solubilizing agents, and the like.

In a composition (such as a food composition or a pharmaceutical composition) that contains a processed product of sardine fruit, the amount of the processed product of sardine fruit varies depending on the aspect thereof, and is adjusted as appropriate. For example, the content (in terms of solid content) of the processed product of the persimmon fruit in the composition containing the processed product of the persimmon fruit is 1% by mass or more, 2% by mass or more, 3% by mass or more, 4% by mass or more, It is preferably 5% by mass or more. Although there is no particular limitation on the upper limit, the upper limit may be set as appropriate, such as when the content (in terms of solid content) of the processed product of Saji fruit is 20% by mass or less, 15% by mass, or the like. In addition, the solid content of the processed product of the candied fruit is the mass of the processed product of the bacillus fruit excluding water and solvent. For example, since the water content of sea bream juice is approximately 80 to 90% by mass, the solid content is approximately 10 to 20% by mass. The content (in terms of solid content) of the processed product of the candied fruit is the value obtained by dividing the solid content of the processed product of the candied fruit by the mass of the composition containing the processed product of the candied fruit, expressed as a percentage.

In addition, when the seaweed fruit juice is a processed product of seaweed fruits, the seaweed juice can be used as it is, or a seaweed juice-containing composition prepared by adding additives to the seaweed juice can be used as an oral composition for promoting erythropoietin production. . The content of Saji fruit juice in the oral composition (mass of Saji fruit juice/mass of oral composition x 100 (%)) is 30% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, It is preferable that the content is higher in the order of 80% by mass or more and 90% by mass or more. Further, the upper limit of the content of the candied fruit juice in the oral composition may be set as appropriate, such as 100% by mass or less, 99.99% by mass or less, 99.98% by mass, or the like.

The intake amount of a composition (food composition, pharmaceutical composition, etc.) containing a processed product of sardine fruit may be appropriately selected depending on the gender, weight, age, purpose of use, etc. of the subject. For example, in the case of an adult human, the daily intake amount of the processed product (in terms of dry solid content) of saji fruit can be 0.1 to 45 g, 1 to 35 g, or 2 to 15 g. In the case of a composition containing seabass fruit juice, the daily intake for an adult human can be, for example, 1 to 300 g, 10 to 250 g, 15 to 200 g, 20 to 100 g, etc. Further, the intake can be carried out once a day or in divided doses, and may be taken continuously over several days, weeks, or months.

EXAMPLES Hereinafter, the present invention will be explained in more detail 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 Saji Juice H. Rhamnoides fruit (pulp and pericarp, produced in Banhanshan, Inner Mongolia) with branches and leaves attached was frozen at -18°C, the branches and leaves were removed while frozen, and washed with drinking water. Removed damaged fruit. Next, the fruit with the pericarp was put into a mixer, pureed, filtered (60 mesh) to remove the pericarp residue and seeds, and subjected to high-temperature instant sterilization to obtain saji juice. Stevia was added to the seaweed juice, mixed while removing foreign substances with a mesh (40 mesh), and heat sterilized to obtain a seaweed juice containing 99.95% by mass of seaweed juice.

II. Measurement of EPO Concentration The freeze-dried product of the soybean 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 bream juice was placed in a freeze dryer and allowed to stand for several days at -50°C and 40Pa to obtain a freeze-dried product. The freeze-dried product of sea radish juice was dissolved in sterilized water at concentrations of 0.02 mg/mL, 0.2 mg/mL, 2 mg/mL, 20 mg/mL, 40 mg/mL, and 80 mg/mL. Thereafter, before adding the sample to a human liver cancer-derived cell line (HepG2 cells), it was diluted 200 times with serum-free EMEM (containing 1% penicillin-streptomycin) (final concentration 0.1 μg/mL, 1 μg/mL, 10 μg/mL). mL, 100 μg/mL, 200 μg/mL, and 400 μg/mL). 100 μL of each 200-fold diluted sample solution was added to the cells.

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

(iii) Control test A control test was conducted in the same manner as in (ii) above, using sterile water instead of the sample solution.

(iv) Evaluation of erythropoietin production amount by ELISA method HepG2 cell culture supernatant was collected 24 hours after addition of the evaluation sample. The amount of EPO produced in the culture supernatant was calculated using the Enzyme-Linked Immunosorbent Assay (ELISA) kit for Erythropoietin (EPO) (Cloud-Clone Crop.). EPO standards, samples and blanks (buffer only without sample) were added to the antibody coated plate. Thereafter, 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 five 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 stop solution was added, and absorbance at 450 nm was measured using a microplate reader.

FIG. 1 shows the erythropoietin concentrations of Example 1 and the control test. As shown in FIG. 1, addition of the freeze-dried sudder juice (200 μg/mL treated 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 group, 10 μg/mL treated group, 100 μg/mL treated group, and 400 μg/mL treated group compared to the control.

Since sea bream juice contains the same components as the freeze-dried sea bream juice other than water, the same effects can be expected with sea bream juice. In addition, the water content contained in the soybean juice obtained in the above I was 86% by mass.

The erythropoietin production promoter of the present invention can be used in foods, medicines, etc., and is industrially useful.

Claims (4)

An erythropoietin production promoter characterized by containing a processed product of sardine fruit as an active ingredient. The erythropoietin production promoter according to claim 1, wherein the processed product of the candied soybean fruit is a candied soybean juice and/or a dried product of the candied soybean juice. The erythropoietin production promoter according to claim 1 or 2, wherein the soybean fruit is a soybean fruit containing pericarp, pulp, and seeds. The erythropoietin production promoter according to claim 1 or 2, which is an oral composition.