JP2023143445A - Uric acid synthesis inhibitor - Google Patents

Abstract

To provide a uric acid synthesis inhibitor.SOLUTION: A uric acid synthesis inhibitor contains at least one selected from the group consisting of Euglena, paramylon, processed paramylon, and β-1,3-glucan.SELECTED DRAWING: None

Description

The present invention relates to uric acid synthesis inhibitors and the like.

It is known that high blood uric acid levels cause various diseases. A blood uric acid level of 7.0 mg/dL or higher is diagnosed as hyperuricemia, and it is said that the risk of developing diseases such as gout, kidney stones, and urinary tract stones is high. Therefore, it is important to suppress the blood uric acid level within a certain value from the viewpoint of disease onset risk. Only 20% to 30% of the purines in the body, which are the source of uric acid production, are ingested through food, and the majority (70% to 80%) is produced within the body. Therefore, by suppressing uric acid synthesis in the body, blood uric acid levels can be effectively suppressed.

Euglena is a microalgae belonging to the genus Euglena (genus Euglena) and is used as a food material. Moreover, applying Euglena extract to the skin has also been practiced (Patent Document 1). Furthermore, paramylon is a β-1,3-glucan produced by Euglena, and has been reported to be useful for wound treatment, allergy suppression, etc. However, the relationship between Euglena and β-1,3-glucan and uric acid synthesis is still unknown.

Special table 2008-526954 publication

An object of the present invention is to provide a uric acid synthesis inhibitor.

As a result of intensive research in view of the above problems, the present inventors have discovered a uric acid synthesis inhibitor containing at least one member selected from the group consisting of Euglena, paramylon, processed paramylon, and β-1,3-glucan; If so, we have found that the above problem can be solved. As a result of further research based on this knowledge, the present invention was completed. That is, the present invention includes the following aspects.

Item 1. A uric acid synthesis inhibitor containing at least one member selected from the group consisting of Euglena, paramylon, processed paramylon, and β-1,3-glucan.

Item 2. Item 2. The uric acid synthesis inhibitor according to item 1, which contains at least one selected from the group consisting of Euglena, paramylon, and processed paramylon.

Item 3. containing Euglena and/or Paramylon, and the Euglena is Euglena gracilis and/or the paramylon is a paramylon derived from Euglena gracilis,
Item 2. The uric acid synthesis inhibitor according to item 1 or 2.

Item 4. contains the Euglena and/or Paramylon, and the Euglena is Euglena gracilis EOD-1 strain (accession number FERM BP-11530), and/or the paramylon is Euglena gracilis EOD-1 strain (Accession number FERM BP- 11530) is a paramylon derived from
Item 2. The uric acid synthesis inhibitor according to item 1 or 2.

Item 5. Item 5. The uric acid synthesis inhibitor according to any one of Items 1 to 4, which contains paramylon.

Item 6. Item 6. The uric acid synthesis inhibitor according to Item 5, wherein the paramylon is purified paramylon.

Section 7. Item 7. The uric acid synthesis inhibitor according to any one of Items 1 to 6, for use in reducing blood uric acid levels by suppressing uric acid synthesis.

Section 8. Suppression of uric acid synthesis according to any one of Items 1 to 7, for use in the prevention or amelioration of at least one selected from the group consisting of hyperuricemia, gout, renal dysfunction, renal stones, and urinary tract stones. agent.

Item 9. Item 9. The uric acid synthesis inhibitor according to any one of Items 1 to 8, for use in inhibiting the uric acid synthesis pathway.

Item 10. Item 10. The uric acid synthesis inhibitor according to any one of items 1 to 9, which is a food composition, a nutritional supplement, a food additive, or a pharmaceutical.

Item 11. Item 11. The uric acid synthesis inhibitor according to any one of Items 1 to 10, which is an oral composition.

According to the present invention, a uric acid synthesis inhibitor can be provided.

2 is a graph showing the measurement results of the expression level of a gene (Nt5e (5 nucleotidase)) in the uric acid synthesis pathway in the ileum of Test Example 1. On the horizontal axis, the control group is a group that ingested a high-fat diet supplemented with lard, and the test group is a group that ingested a high-fat diet supplemented with paramylon instead of cellulose contained in the high-fat diet. be. The vertical axis shows the relative signal of the gene expression level (mRNA level) when the signal of the control group is set to 1. 2 is a graph showing the measurement results of the expression level of a gene (Ada (adenosine deaminase)) in the uric acid synthesis pathway in the ileum of Test Example 1. The explanation of the horizontal and vertical axes is the same as in FIG. 2 is a graph showing the measurement results of the expression level of a gene of the uric acid synthesis pathway (Pnp2 (purine-nucleoside phosphorylase 2)) in the liver of Test Example 1. The explanation of the horizontal and vertical axes is the same as in FIG. 3 is a graph showing the measurement results of uric acid levels in Test Example 1. On the horizontal axis, the control group is a group that ingested a high-fat diet supplemented with lard, and the test group is a group that ingested a high-fat diet supplemented with paramylon instead of cellulose contained in the high-fat diet. be. The vertical axis shows the uric acid value. * indicates that the p value is less than 0.05 when compared with the control group by t-test.

In this specification, the expressions "contain" and "including" include the concepts of "containing", "comprising", "consisting essentially" and "consisting only".

In one aspect, the present invention provides a uric acid synthesis inhibitor (herein referred to as " The agent of the present invention may also be referred to as the agent of the present invention. This will be explained below.

1. Euglena Euglena is a microalgae belonging to the genus Euglena (=genus Euglena), and is not particularly limited as far as it is concerned. Specifically, examples of Euglena include Euglena gracilis , Euglena longa , Euglena caudata , Euglena oxyuris , Euglena tripteris , Euglena proxima , Euglena viridis , Euglena sociabilis , Euglena ehrenbergii , Euglena deses , Euglena pisciformis , Euglena spirogyra , Euglena acus , Euglena geniculata , Euglena intermedia , Euglena mutabilis , Euglena sanguinea , Euglena stellata , Euglena terricola , Euglena klebsi , Euglena rubra , Euglena cyclopicola and the like. Among these, Euglena gracilis is preferred from the viewpoint of more reliably exerting the effects of the present invention, and more preferably Euglena gracilis EOD-1 strain [Independent Administrative Agency Product Evaluation as of June 28, 2013] The National Institute of Technology and Technology, Patent Organism Depositary Center {NITE-IPOD (Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818)} under the provisions of the Budapest Treaty, with accession number FERM BP-11530. Internationally deposited].

The form of Euglena is not particularly limited as long as it contains the cell body of Euglena or most of its components. Examples of the form of Euglena include a dry powder form of Euglena, a suspension of Euglena, and an extract of Euglena, and among them, a dry powder form of Euglena is preferred.

The paramylon content of Euglena in a dry state is, for example, 50% or more, preferably 60% or more, and more preferably 70% or more.

Euglena may be used alone or in combination of two or more.

2. β-1,3-glucan, paramylon β-1,3-glucan is a substance that has one sugar chain (or sugar chain structure) consisting of glucose linked only by β1,3 bonds as its main chain. There are no particular restrictions. β-1,3-glucans are not limited to linear ones, but also include those with branched chains.

The weight average molecular weight of the β-1,3-glucan derivative is not particularly limited, but is, for example, 1×10 ⁴ to 2×10 ⁶ , preferably 5×10 ⁴ to 1×10 ⁶ , more preferably 1×10 ⁵ to There are 1×10 ⁶ . Note that the weight average molecular weight can be measured by the GPC method.

β-1,3-glucan may be obtained by chemical synthesis, but natural β-1,3-glucan produced by various organisms is preferable from the viewpoint of availability. Examples of natural β-1,3-glucans include paramylon, curdlan, laminaran, callose, lentinan, and schizophyllan. Among these, paramylon is particularly preferred. Paramylon will be explained below.

Paramylon is β-1,3-glucan derived from Euglena, and is not particularly limited.

Regarding Euglena, from which Paramylon is derived, the explanation is the same as in "1. Euglena" above.

The mass average molecular weight of paramylon is not particularly limited, but is, for example, 1×10 ⁴ to 5×10 ⁶ , preferably 2×10 ⁴ to 1×10 ⁶ , more preferably 5×10 ⁴ to 1×10 ⁶ , even more preferably is 1×10 ⁵ to 5×10 ⁵ .

The mass average molecular weight can be measured by SEC-MALS analysis under the following conditions:
Detector: Multi-angle scattering detector (DAWN HELEOS II manufactured by Wyatt Technology)
Differential refractometer detector (Optilab T-rEX manufactured by Wyatt Technology)
Columns used: 2 TSKgel α-M (manufactured by Tosoh)
Mobile phase: DMSO with 0.05M potassium bromide
Flow rate: 0.5 mL/min.

Paramylon normally exists in Euglena cells as paramylon particles, which are highly regularized triple helical structures formed by β-1,3-glucan chains.

Although the shape of the paramylon particles is not particularly limited, it is usually a flat spheroid shape.

The particle size distribution of paramylon particles is not particularly limited, but is, for example, 0.5 to 15 μm, preferably 1 to 6 μm. Further, the average particle diameter of the paramylon particles is also not particularly limited, but is, for example, 1 to 10 μm, preferably 2 to 4 μm.

The form of paramylon is not particularly limited as long as it contains paramylon. Examples of the form of Euglena include a dry powder form of paramylon, a suspension of paramylon, and the like, and among them, a dry powder form of paramylon is preferable.

In one embodiment, the agent of the present invention preferably contains purified paramylon.

Paramylon may be used alone or in combination of two or more.

3. Method for producing Euglena and Paramylon Euglena can be prepared in large quantities by a method that includes a step of culturing Euglena contained in a liquid (culture step). The culturing step can be performed, for example, according to a known method (for example, the method described in Japanese Patent No. 5883532). In the culturing step, microalgae belonging to the genus Euglena are typically cultured under aerobic conditions while stirring a liquid (culture solution) containing water, Euglena, and nutrients available to Euglena.

Nutrients include sugars (monosaccharides such as glucose and fructose), minerals (such as sodium, potassium, magnesium, calcium, iron, zinc, molybdenum, copper, phosphorus, nitrogen, sulfur, or boron). ), B vitamins (e.g. vitamin B1 (thiamine), vitamin B2 (riboflavin), niacin, pantothenic acid, vitamin B6 (pyridoxine, pyridoxal, or pyridoxamine), vitamin B12 (cyanocobalamin), folic acid, biotin, etc.). It will be done. The concentration of nutrients in the culture solution is not particularly limited as long as the concentration allows survival, proliferation, etc. of Euglena.

The light conditions for the culturing process are not particularly limited, and the culturing process may be performed under either bright or dark conditions. When culturing by heterotrophic culture, it is cultured under dark conditions. As the light condition, a normal light intensity for growing algae can be adopted. The dark conditions include, for example, less than 10 μmol/m ² /s, preferably complete dark conditions with no light.

The culture temperature in the culture step is not particularly limited as long as it is a temperature at which Euglena can proliferate. The culture temperature (temperature of the culture solution) is, for example, 20°C to 35°C.

The pH of the liquid in the culture step is not particularly limited as long as it allows Euglena to proliferate. The pH at which Euglena can grow is, for example, 3.0 to 5.5.

After the culturing step, it is preferable to concentrate Euglena by liquid centrifugation, gravity separation, or the like. The obtained Euglena can be subjected to additional treatments (eg, suspension in liquid, dispersion in water or oil, extraction, dry powdering, etc.) depending on the desired form.

Paramylon particles can be produced by separating, isolating, or purifying Euglena from Euglena according to or in accordance with a known method (for example, the method described in Japanese Patent No. 5883532). Paramylon particles can be easily obtained, for example, by recovering cellular components obtained by disrupting the cell membrane of Euglena. Furthermore, the paramylon particles may be purified if necessary. Various methods are known for purifying paramylon particles (for example, Japanese Patent No. 5883532), and purification can be performed according to those methods. Examples of the purification process include a surfactant treatment process and a washing process. The obtained Euglena can be subjected to additional treatments (eg, suspension in liquid, dispersion in water or oil, dry powdering, etc.) depending on the desired form.

Four. Paramylon-processed product Paramylon-processed product is obtained by processing paramylon, for example, physical treatment, chemical treatment, etc., and is not particularly limited as far as it is concerned. Examples of paramylon processed products include fibrous paramylon, amorphous paramylon, and the like. Amorphous paramylon can be obtained by chemical treatment according to or analogously to a known method, for example, using the method described in JP-A No. 2011-184592.

As the paramylon processed product, fibrous paramylon is preferable. The fiberized paramylon will be explained below.

Fibrillated paramylon is β-1,3-glucan derived from Euglena, and is not particularly limited as long as it is in a fibrous form. Until now, amorphous paramylon obtained by chemically treating paramylon particles (alkali treatment, etc.) has been reported, but when observed with an electron microscope, it was not recognized that it was fibrous, and the shape and size were Since it is an amorphous lump, it is not included in fibrous paramylon.

The weight average molecular weight of the fibrous paramylon is not particularly limited, but is, for example, 1×10 ⁴ to 2×10 ⁷ , preferably 1×10 ⁵ to 5×10 ⁵ .

In addition, the weight average molecular weight can be measured by the following method by SEC-MALS analysis:
Detector: Multi-angle scattering detector (DAWN HELEOS II manufactured by Wyatt Technology)
Differential refractometer detector (Optilab T-rEX manufactured by Wyatt Technology)
Columns used: 2 TSKgel α-M (manufactured by Tosoh)
Mobile phase: DMSO with 0.05M potassium bromide
Flow rate: 0.5 mL/min.

The fiber diameter of the fiberized paramylon is not particularly limited, but is, for example, 10 to 500 nm, preferably 20 to 300 nm, more preferably 50 to 200 nm. The fiber diameter of fibrous paramylon can usually be measured based on an electron microscope image of fibrous paramylon.

The settling volume of the fibrous paramylon in water is not particularly limited, but is, for example, 30 to 300 mL/g, preferably 50 to 250 mL/g, and more preferably 70 to 200 mL/g.

The submerged volume can be measured according to or analogously to the following method:
Measurement is carried out according to the method described in "Dietary Fiber - Basics and Applications - 3rd edition, p. 111, Daiichi Publishing, Tokyo" (2008), supervised by the Japanese Dietary Fiber Society, edited by the Editorial Committee of the Japanese Dietary Fiber Society. . Specifically, it is as follows. Weigh 125 mg of the sample slurry test sample into a 25 mL plastic tube in terms of dry mass, and shake the plastic tube vigorously by hand to stir the contents. Then, transfer the contents to a 25 mL graduated cylinder and add pure water until the volume reaches 25 mL. After stirring the liquid in the graduated cylinder, let it stand at 37°C for 24 hours. This causes the sample to precipitate, resulting in two layers that are separated via an interface: a layer containing mainly the precipitated sample (lower layer), and a layer containing mainly water (upper layer). Determine the volume of the lower layer from the scale of the graduated cylinder, and divide the obtained volume by the sample mass (dry mass) to calculate the submerged volume (mL/g). The test is performed 3 or 4 times, and the average value and standard deviation are calculated.

Fibrillated paramylon has a relatively high resistance to enzymatic degradation. For example, the amount of monomer (glucose) produced by the degradation of β-glucanase is, for example, 0.1 to 50 mg, preferably 1 to 10 mg, per gram of fibrillated paramylon.

This amount can be measured according to or analogously to the following method:
Reaction solution [test substance 30 mg (dry weight), buffer solution (Tokyo Kasei Kogyo Co., Ltd. B0156, potassium hydrogen phthalate-sodium hydroxide buffer (pH4.0)) 5 mL, enzyme solution (Nippon Biocon Co., Ltd. endo- Prepare 0.1 mL of 1,3-β-Glucanase (enzyme content: 50 units/mL), pure water, reaction volume 10 mL] and shake horizontally at 45 rpm at 40°C for 24 hours. After shaking, immediately store frozen and lyophilize for concentration. After freeze-drying, add 0.5 mL of pure water to each sample and stir (concentrate 20 times). Repeat the process of centrifuging (10,000G, 5 minutes, 4℃) and collecting the supernatant twice. The glucose concentration in the collected supernatant is measured using a measurement kit (Glucose CII-Test Wako, manufactured by Wako Pure Chemical Industries, Ltd.). Based on the measured values, calculate the amount of glucose produced (mg) per 1 g of the test substance.

Fiberized paramylon has relatively low solubility in alkaline solutions. For example, fibrous paramylon does not dissolve in a 0.1-0.3M aqueous sodium hydroxide solution. Here, "does not dissolve" means, for example, that the absorbance (660 nm) of the solution after suspending the fibrous paramylon in the aqueous solution (for example, immediately after to 1 hour) is, for example, 0.1 or more, preferably 1.0. It means the above.

Solubility can be measured according to or analogously to the following method:
Suspend 250 mg (dry weight) of the test substance in 10 mL of test solution (pure water, 0.1M NaOH aqueous solution, 0.3M NaOH aqueous solution) in a vial. The absorbance of the liquid in the vial is measured at 660 nm after shaking the vial vigorously by hand for 20 seconds and after shaking for 1 hour at 80 rpm on a shaker, respectively. The absorbance is measured using a spectrophotometer V-730 manufactured by JASCO Corporation.

The relative value of the crystallinity of fibrous paramylon to granular paramylon (crystallinity of fibrous paramylon/crystallinity of granular paramylon) is, for example, 0.60 to 0.90, preferably 0.65 to 0.80.

Crystallinity can be measured according to or analogously to the following method:
Perform XRD measurements on the test substance. The conditions are as follows. Equipment: PANalytical X'Pert3 Powder, tube voltage: 45kV, tube current: 40mA, measurement range: 5.005 to 50.018°, measurement interval: 0.013°, analysis software: HighScore. The degree of crystallinity is analyzed by the ratio of the strength of the amorphous part to the strength of the crystalline part at 2θ=5 to 80°. The analysis was performed after removing the background caused by the device from each measurement (background setting Auto, venting factor 0, granularity 100), and the amorphous part was determined by the tangent line passing through 2θ = 14 and 29°. do. The pending factor and granularity conditions that determine each amorphous part are 0/20.

The fibrous paramylon may be in a form dispersed in a solvent such as water, or may be in a dried form. Fiberized paramylon, even in dry form, can be redispersed in water.

In addition, in this specification, "dry form" indicates that the moisture content is 15% by mass or less, preferably 10% by mass or less, and more preferably 5% by mass or less.

As the fiberized paramylon, it is preferable to use a defibrated product of paramylon particles obtained by physically defibrating the paramylon particles. Further, a defibrated product of Euglena obtained by applying this defibration treatment to Euglena can also be used as fiberized paramylon.

The defibration treatment hardly breaks the hydrogen bonds of β-1,3 glucan present in paramylon particles (for example, 10% or less, 5% or less, 2% or less of the hydrogen bonds of β-1,3 glucan). , a treatment that can defibrate (with less than 1% cleavage), or unravel part or all of the β-1,3-glucan chains present in the paramylon particles or the triple helical structure formed by the β-1,3-glucan chains. There is no particular restriction as long as the processing can be performed. Preferably, it is preferable to defibrate the β-1,3 glucan present in the paramylon particles without breaking most of the hydrogen bonds, thereby forming the paramylon particles into a fibrous form. Any known process capable of grinding (shearing) or pulverizing (preferably grinding (shearing)) fine particles such as paramylon particles can be employed as the fibrillation process.

The defibration treatment can be performed using a known device such as a mill (shear) or a crusher. Examples of devices used for the fibrillation treatment include stone mill mills, jet mills, twin-screw kneaders, high-pressure homogenizers, high-pressure emulsifiers, twin-screw extruders, and bead mills. Among these, stone mill type mills and bead mills are preferred.

The defibration treatment can be performed wet or dry. It is preferable to perform the defibration treatment in a wet manner because it allows the fiberized paramylon to be more efficiently dispersed in the solution. The solvent used in the wet process is not particularly limited as long as it is a solvent in which the fibrous paramylon can be dispersed, and water can be suitably used.

The defibration treatment may be carried out by one type alone or by a combination of two or more types. Further, a part of the paramylon may be defibrated, and as long as the paramylon includes defibrated paramylon, it is within the scope of the present invention.

Five. Uses At least one selected from the group consisting of Euglena, paramylon, paramylon processed products, and β-1,3-glucan (hereinafter sometimes referred to as "active ingredient of the present invention") has a uric acid synthesis inhibitory effect. Therefore, it can be used as an active ingredient of a uric acid synthesis inhibitor.

"Suppression of uric acid synthesis" means to suppress the synthesis of uric acid itself in the body, or in other words, to suppress the ability to synthesize uric acid in the body.

Since the active ingredient of the present invention has a blood uric acid level reducing effect, it can be used as an active ingredient of a blood uric acid level reducing agent, particularly an agent for reducing blood uric acid level by inhibiting uric acid synthesis. Furthermore, in one embodiment of the present invention, the agent of the present invention can be used to suppress uric acid synthesis and reduce blood uric acid levels.

Since the active ingredient of the present invention has a uric acid synthesis pathway inhibitory effect, it can be used as an active ingredient of a uric acid synthesis pathway inhibitor. Furthermore, in one embodiment of the present invention, the agent of the present invention can be used for inhibiting the uric acid synthesis pathway.

"Suppression of uric acid synthesis pathway" refers to suppressing the function of enzymes in the uric acid synthesis pathway. Examples of enzymes in the uric acid synthesis pathway include 5 nucleotidase (an enzyme responsible for converting adenylate to adenosine and guanylate to guanosine in the uric acid synthesis pathway), adenosine deaminase (an enzyme responsible for converting adenosine to inosine in the uric acid synthesis pathway), Purine-nucleoside phosphorylase 2 (an enzyme responsible for converting inosine to hypoxanthine and guanosine to guanine in the uric acid synthesis pathway), etc.

Based on the above-mentioned effects, the active ingredient of the present invention can be used for the prevention and improvement of hyperuricemia, gout, renal dysfunction, renal stones, urinary tract stones, and the like.

The active ingredient of the present invention preferably has multiple uses (two or more, more preferably three or more, even more preferably four or more, even more preferably five or more, even more preferably six) of the above uses. It can be used for a comprehensive range of applications, including those listed above.

Note that "suppression" includes not only reducing, but also suppressing the increase or deterioration of something that is increasing or deteriorating (reducing the width of increase or the degree of deterioration, or preventing increase or deterioration).

Furthermore, the active ingredient of the present invention has the following uses, purposes, and objects:
(a) Lowers uric acid levels (b) Suppresses increases in uric acid levels (c) Suppresses uric acid synthesis and production (d) Suitable for people with high uric acid levels (e) Supports uric acid (f) Uric acid It can also be used for value care, etc.

The agent of the present invention is applicable to those with blood uric acid levels in the normal range (5.5 mg/dL or more and 7.0 mg/dL or less) and high values (7.1 mg/dL or more), or those with blood uric acid levels in the normal range. It is preferable that the blood uric acid level is high (5.5 mg/dL or more and 7.0 mg/dL or less) and mild (7.1 mg/dL or more and 7.9 mg/dL or less), and the blood uric acid level is high in the normal range (5.5 mg/dL or more). 7.0 mg/dL or less) is particularly preferable. By applying the agent of the present invention to such subjects, it is possible to more effectively suppress uric acid synthesis and reduce blood uric acid levels.

The subject to whom the agent of the present invention is applied is preferably obese. For example, the subject can have a BMI of, for example, 24 or higher, preferably 25 or higher, and more preferably 25 to 30. By applying the agent of the present invention to such subjects, it is possible to more effectively suppress uric acid synthesis and reduce blood uric acid levels.

The agent of the present invention can be used in various fields, such as food compositions (including health foods, health promoters, nutritional supplements (supplements, etc.)), food additives, cosmetics, cosmetic additives, medicines, reagents, feeds, etc. It can be used as The agent of the present invention is preferably an oral composition.

The form of the agent of the present invention is not particularly limited, and depending on the application, it can take any form commonly used in each application.

When the agent of the present invention is used as a food composition, liquid, gel, or solid foods such as juice, soft drinks, tea, soup, soy milk, salad oil, dressing, yogurt, jelly, etc. , pudding, furikake, powdered milk for infants, cake mixes, dairy products (e.g., powder, liquid, gel, solid, etc.), bread, confectionery (e.g., cookies, etc.), and the like.

When the agent of the present invention is used as a cosmetic, for example, emulsion, lotion, face cream, hand cream, lotion, body soap, shampoo, conditioner, cosmetic gel, pack, foundation, lip balm, face wash, etc. agents, etc.

When the agent of the present invention is used as a medicine, examples include ointments, external liquids (liniments, lotions, etc.), sprays (external aerosols, pump sprays, etc.), creams, and gels. , patches (plasters, plasters, tapes (reservoir type, matrix type, etc.), poultices, patches, microneedles, etc.), eye drops, eye ointments, nasal drops, suppositories, rectal semi-prescription Pharmaceutical forms suitable for parenteral ingestion such as solid preparations and enema preparations (especially external preparation forms); Tablets (including orally disintegrating tablets, chewable tablets, effervescent tablets, troches, jelly drops, etc.); Pharmaceutical forms suitable for oral ingestion such as pills, granules, fine granules, powders, hard capsules, soft capsules, dry syrups, liquids (including drinks, suspensions, and syrups), and jellies ( oral preparation form).

The form of the agent of the present invention may be, for example, a tablet (orally disintegrating tablet, chewable tablet, effervescent tablet, troche, jelly, etc.) when the application is an additive, a health promoter, a nutritional supplement, etc. (including drop formulations), pills, granules, fine granules, powders, hard capsules, soft capsules, dry syrups, liquid formulations (including suspensions and syrups), and jelly formulations.

The agent of the present invention may further contain other components as necessary. Other ingredients include ingredients that can be incorporated into food compositions (including health foods, health promoters, nutritional supplements (supplements, etc.)), food additives, cosmetics, cosmetic additives, medicines, reagents, feeds, etc. Examples include bases, carriers, solvents, dispersants, emulsifiers, buffers, stabilizers, excipients, binders, disintegrants, lubricants, thickeners, coloring agents, fragrances, chelating agents, etc.

The content of the active ingredient in the agent of the present invention depends on the purpose, mode of use, condition of the subject, etc., and is not limited to, for example, 0.0001 to 100% by mass, preferably 0.001 to 50% by mass. can do.

The amount of the agent of the present invention to be applied (for example, administration, ingestion, inoculation, etc.) is not particularly limited as long as it is an effective amount that exhibits the uric acid synthesis inhibitory effect, and is generally expressed as the dry weight of the active ingredient per day. 0.1-10000 mg/kg body weight. It is preferable to apply the above-mentioned amount in divided doses at least once a day (for example, 1 to 3 times), and it can be increased or decreased as appropriate depending on age, pathological condition, and symptoms.

In a preferred embodiment of the present invention, the application amount (dry weight) of Euglena or Paramylon is preferably 100 to 10,000 mg, more preferably 1,000 to 7,000 mg, and even more preferably 2,000 to 5,000 mg per day. The application period is preferably 1 week or more, more preferably 4 weeks or more, and even more preferably 8 weeks or more, and it is also possible to apply for a longer period of time (10 weeks or more, 15 weeks or more, or 20 weeks or more). . Since the active ingredient of the present invention is naturally derived and highly safe, the upper limit of the application period is not particularly limited, and is, for example, 3 years, 1 year, 6 months, or 4 months.

The present invention will be described in detail below based on Examples, but the present invention is not limited to these Examples.

Reference example 1
As Euglena, dry powder of Euglena gracilis EOD-1 strain (National Institute of Technology and Evaluation, Patent Organism Depositary (NITE-IPOD) (manufactured by Kobelco Eco-Solutions, paramylon content of 70% or more) was prepared.

Reference example 2
Paramylon particles were prepared as follows.

The prepared Euglena gracilis EOD-1 strain (after culture but before drying) was collected in 5 flasks of liquid, and the collected liquid was centrifuged in a centrifuge tube (500 x g, 4 minutes, room temperature). . The supernatant in the centrifuge tube was once removed and collected. The collected supernatant liquid was placed in a centrifuge tube to disperse the precipitate in the centrifuge tube, and the entire solution was transferred to a 100 mL graduated cylinder. Furthermore, the collected supernatant liquid was added to the graduated cylinder to make up the volume to 90 mL.

[Enzyme treatment process]
The liquid was diluted to 90 mL and transferred to a 200 mL beaker, and the pH of the liquid was adjusted to 3 by adding an aqueous hydrochloric acid solution while stirring. A proteolytic enzyme (acidic protease, product name "Protease YP-SS" manufactured by Yakult Pharmaceutical Co., Ltd., optimum pH 2.5 to 3.0) was added to the liquid at a concentration of 5 g/L. Enzyme treatment was performed at 50°C for 2 hours while stirring the liquid.

[Surfactant treatment process]
An aqueous solution of sodium dodecyl sulfate was added to the liquid that had undergone the enzyme treatment step so that the concentration of sodium dodecyl sulfate was 3.0% by mass/volume (w/v). While stirring the liquid containing sodium dodecyl sulfate, the pH of the liquid was adjusted to 3 by adding an aqueous hydrochloric acid solution. Furthermore, the liquid was stirred for 30 minutes at 60°C with a propeller stirrer (rotation speed 200 rpm).

[Separation process]
Paramylon was precipitated by centrifugation (1000 x g, 2 minutes, room temperature), and paramylon was separated from the liquid that had undergone the surfactant treatment step. A further surfactant treatment step was carried out in the same manner except that the concentration of sodium dodecyl sulfate was changed to 1.0% by mass/volume and the pH was not adjusted. Thereafter, a separation step was performed in the same manner as above. In this way, the surfactant treatment step and separation step were each performed three times.

[Washing process]
Paramylon precipitated by centrifugation in the separation step was suspended in pure water and allowed to stand at 40°C for 10 minutes. Paramylon was then precipitated by centrifugation (1000 x g, 2 min, room temperature). Such operations were performed three times in total.

[Drying process]
Paramylon precipitated by centrifugation in the washing step was dried at 50°C to obtain paramylon particles. The obtained paramylon particles were used as paramylon (purified paramylon) in the following test examples.

Test example 1. Analysis of the effect on the gene expression level of the uric acid synthesis pathway Mice were raised with feed containing paramylon (Reference Example 2), and the gene expression level of the uric acid synthesis pathway was measured. Specifically, it was performed as follows.

<1-1. Test method>
<1-1-1. Experimental animals and breeding conditions>
Four-week-old male C57BL/6J mice (manufactured by Charles River Japan) were used. After preliminarily rearing for one week on solid feed (NMF, manufactured by Oriental Yeast Industry Co., Ltd.), the mice were divided into two groups of 8 mice per group so that the weight was uniform.

The feed used in the test is as follows. 20% lard was added to the feeds of the control group and the test group so that the fat/energy ratio was 50%. Cellulose was added to the control group's feed so that the dietary fiber weight percentage was 5%, and paramylon was added to the test group feed so that the dietary fiber weight percentage was 5%. Table 1 shows the feed composition of each group. The values in Table 1 are the weight (g) of each component in 1 kg of feed.

In the test, mice were given ad libitum access to the above feed and water for 12 weeks. The breeding environment was a temperature of 22±1°C, a humidity of 50±5%, and a 12-hour light/dark cycle (light period: 9:00 to 21:00, dark period: 21:00 to 9:00). On the final day of the test, the animals were fasted for 8 hours and euthanized with isoflurane/carbon dioxide gas. Blood was collected and serum was separated. In addition, the ileum and liver were removed and stored in RNA later (Qiagen) and used as samples for RNA extraction.

<1-1-2. Measurement of gene expression level of uric acid synthesis pathway>
After extracting RNA from the ileum and liver using the RNeasy mini kit (Qiagen), the expression level of gene mRNA for the uric acid synthesis pathway was measured using a microarray method (array: SurePrint G3 Mouse GE 8x60K (Agilent), equipment: GeneChi Scanner 3000 7G System (Thermo Fisher Scientific), analysis software: Transcriptome Viewer (Kurabo Industries)). The genes of the uric acid synthesis pathway that were measured are as follows.
・Nt5e (5 nucleotidase, Gene Accession: NM_011851)
・Ada (adenosine deaminase, Gene Accession: NM_001272052)
・Pnp2 (purine-nucleoside phosphorylase 2, Gene Accession: NM_001123371)
<1-1-3. Measurement of uric acid level>
Serum uric acid levels were measured by an enzymatic method (uricase/TOOS method).

<1-2. Results＞
The measurement results of gene expression levels of the uric acid synthesis pathway are shown in FIGS. 1 to 3. 5 Nucleotidase is an enzyme responsible for converting adenylate to adenosine and guanylate to guanosine in the uric acid synthesis pathway, and adenosine deaminase is an enzyme responsible for converting adenosine to inosine in the uric acid synthesis pathway, and purine-nucleoside Phosphorylase 2 is an enzyme responsible for converting inosine to hypoxanthine and guanosine to guanine in the uric acid synthesis pathway. Hypoxanthine and guanine are precursors that are converted to uric acid. As shown in FIGS. 1 to 3, paramylon intake suppressed the expression of these genes, indicating that paramylon has an inhibitory effect on the uric acid synthesis pathway and an inhibitory effect on uric acid synthesis.

The measurement results of uric acid levels are shown in Figure 4.

Manufacturing example 1
Dry powder of Euglena gracilis EOD-1 strain (National Institute of Technology and Evaluation, Patent Organism Depositary (NITE-IPOD) (manufactured by Kobelco Eco-Solutions, paramylon content of 70% or more)) is made into capsule tablets (capsule composition) with the following composition. Ingredients: pullulan and kolyan pigment) were used as the test food, and capsule tablets containing cellulose (capsule constituents: pullulan and kolyan pigment) were manufactured as a control food.

Test example 2. The outline of the analysis study for the effect on blood uric acid levels is as follows.
・Test food intake period: 12 weeks ・Test food intake: 15 capsules/day (5 capsules each before breakfast, lunch, and dinner)
- Subjects: 9 subjects The subjects had blood uric acid levels (uric acid levels in serum) of 5.5 mg/dL or more and 7.0 mg/dL or less at the start of the study.

There were two types of test food (manufacturing example 1): test food and control food (placebo). The test food was a food containing Euglena gracilis EOD-1 strain, and the control food (placebo) was a food not containing Euglena gracilis EOD-1 strain. .

The subjects ingested 15 capsules of the test food a day (three times a day: 5 capsules each before breakfast, lunch, and dinner) for 12 weeks. The total content of ingredients in the 15 capsules is as follows: Test food: Contains 3.75g of Euglena gracilis EOD-1 strain (contains 2.6g as β-glucan). Control food: Contains 2.6g of cellulose.

At the end of the study (12 weeks after the start of intake), blood was collected from each subject. Serum was separated from the collected blood, and the uric acid level in the serum was measured according to a conventional method.

Serum uric acid levels were compared. For statistical analysis, an analysis of covariance was performed using the measured values at the start of the study (week 0) as a covariate, and the measured values at 12 weeks in both groups were compared. The results are shown in Table 3.

As shown in Table 3, it was found that blood uric acid levels could be reduced by ingesting paramylon.

Claims (11)

A uric acid synthesis inhibitor containing at least one member selected from the group consisting of Euglena, paramylon, processed paramylon, and β-1,3-glucan. The uric acid synthesis inhibitor according to claim 1, containing at least one selected from the group consisting of Euglena, paramylon, and processed products of paramylon. containing Euglena and/or Paramylon, and the Euglena is Euglena gracilis and/or the paramylon is a paramylon derived from Euglena gracilis,
The uric acid synthesis inhibitor according to claim 1 or 2. contains the Euglena and/or Paramylon, and the Euglena is Euglena gracilis EOD-1 strain (accession number FERM BP-11530), and/or the paramylon is Euglena gracilis EOD-1 strain (Accession number FERM BP- 11530) is a paramylon derived from
The uric acid synthesis inhibitor according to claim 1 or 2. The uric acid synthesis inhibitor according to any one of claims 1 to 4, which contains paramylon. The uric acid synthesis inhibitor according to claim 5, wherein the paramylon is purified paramylon. The uric acid synthesis inhibitor according to any one of claims 1 to 6, which is used to suppress uric acid synthesis and reduce blood uric acid levels. Uric acid synthesis according to any one of claims 1 to 7, for use in the prevention or amelioration of at least one selected from the group consisting of hyperuricemia, gout, renal dysfunction, renal stones, and urinary tract stones. Suppressant. The uric acid synthesis inhibitor according to any one of claims 1 to 8, for use in inhibiting the uric acid synthesis pathway. The uric acid synthesis inhibitor according to any one of claims 1 to 9, which is a food composition, a nutritional supplement, a food additive, or a medicine. The uric acid synthesis inhibitor according to any one of claims 1 to 10, which is an oral composition.

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