WO2009113512A1 - Promoteur d'expression de socs3, médicament et aliment contenant celui-ci et procédé de promotion de l'expression de socs3 - Google Patents

Promoteur d'expression de socs3, médicament et aliment contenant celui-ci et procédé de promotion de l'expression de socs3 Download PDF

Info

Publication number
WO2009113512A1
WO2009113512A1 PCT/JP2009/054491 JP2009054491W WO2009113512A1 WO 2009113512 A1 WO2009113512 A1 WO 2009113512A1 JP 2009054491 W JP2009054491 W JP 2009054491W WO 2009113512 A1 WO2009113512 A1 WO 2009113512A1
Authority
WO
WIPO (PCT)
Prior art keywords
hyaluronic acid
expression
socs3
molecular weight
salt
Prior art date
Application number
PCT/JP2009/054491
Other languages
English (en)
Japanese (ja)
Inventor
智行 金光
稔秀 佐藤
晃 浅利
Original Assignee
キユーピー株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by キユーピー株式会社 filed Critical キユーピー株式会社
Priority to JP2010502813A priority Critical patent/JPWO2009113512A1/ja
Publication of WO2009113512A1 publication Critical patent/WO2009113512A1/fr

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
    • A61K31/728—Hyaluronic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L5/00—Compositions of polysaccharides or of their derivatives not provided for in groups C08L1/00 or C08L3/00
    • C08L5/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof

Definitions

  • the present invention relates to a SOCS3 expression promoter, pharmaceuticals and foods containing the same, and a method for promoting the expression of SOCS3.
  • the present invention also relates to a proiotrophin expression inhibitor, pharmaceuticals and foods containing the same, and a method for suppressing the expression of proiotrophin.
  • An autoimmune disease is an immune system that is originally responsible for recognizing and eliminating foreign substances, such as bacteria, viruses, and tumors, and reacts excessively to its own normal cells and tissues. Is a general term for diseases that cause symptoms, and this is caused by abnormalities in immunocompetent cells such as T cells, B cells, and macrophages.
  • WO2002-074318 describes that high molecular weight hyaluronic acid suppresses the expression of interleukin 12 that promotes inflammation.
  • Interleukin 12 is one of the cytokines that promote inflammation in autoimmune diseases.
  • autoimmune diseases develop various symptoms at various sites, it is difficult to comprehensively improve immune diseases even if only a single cytokine is controlled.
  • autoimmune arthritis is one of the autoimmune diseases.
  • Suppressor of cytokine signaling 3 (SOCS3) which regulates signal transduction through cytokines, is an autoimmune arthritis.
  • SOCS3 is said to be strongly expressed in chronic inflammatory diseases such as human colitis and arthritis (Hiromi Takagi, Takato Sanada, Yasumasa Hamada, Akihiko Yoshimura, “Control of cytokines and Toll-like receptor signals by SOCS”, Japan Lin, Vol. 62, No. 12 (2004-12), 2189-2196).
  • SOCS3 has been reported to indirectly regulate Toll-like receptors in innate immune cells (Andrea Baetz, Markus Frey, Klaus Heeg, and Alexander H. Dalpke, “Suppressor of Cytokine Signaling (SOCS) Proteins Indirectly Regulate Toll-like Receptor Signaling in Innate Immune Cells”, J. Biol. Chem., Vol. 279, Issue 52, 54708-54715, December 24.
  • the present invention provides a SOCS3 expression promoter that can comprehensively improve inflammatory diseases, pharmaceuticals and foods containing the same, and a method for promoting the expression of SOCS3.
  • the present invention also provides a proiotrophin expression inhibitor capable of comprehensively improving inflammatory diseases, pharmaceuticals and foods containing the same, and a method for suppressing the expression of proiotrophin. To do.
  • the SOCS3 expression promoter according to one embodiment of the present invention contains hyaluronic acid having an average molecular weight of 500,000 or more and / or a salt thereof as an active ingredient.
  • the pharmaceutical product according to one embodiment of the present invention contains the above-described SOCS3 expression promoter as an active ingredient.
  • the average particle size of the hyaluronic acid and / or salt thereof may be 50 to 500 ⁇ m.
  • the pharmaceutical product can be taken orally. Further, in this case, the pharmaceutical product can be used for treating knee joint or alleviating knee joint pain.
  • the food according to one embodiment of the present invention contains the above SOCS3 expression promoter.
  • a method for promoting the expression of SOCS3 in a human or non-human animal allows a human or non-human animal to orally ingest hyaluronic acid and / or a salt thereof having an average molecular weight of 500,000 or more. Including that.
  • the proiotrophin expression inhibitor according to one embodiment of the present invention contains hyaluronic acid having an average molecular weight of 500,000 or more and / or a salt thereof as an active ingredient.
  • the pharmaceutical product according to one embodiment of the present invention contains the pleiotrophin expression inhibitor as an active ingredient.
  • the average particle size of the hyaluronic acid and / or salt thereof may be 50 to 500 ⁇ m.
  • the pharmaceutical product can be taken orally. Further, in this case, the pharmaceutical product can be used for treating knee joint or alleviating knee joint pain.
  • the food according to one embodiment of the present invention contains the pleiotrophin expression inhibitor.
  • a method for suppressing the expression of proiotrophin in a human or non-human animal includes orally administering hyaluronic acid and / or a salt thereof having an average molecular weight of 500,000 or more to a human or non-human animal. Ingestion.
  • a method for treating a knee joint in a human or non-human animal is to allow a human or non-human animal to orally ingest hyaluronic acid and / or a salt thereof having an average molecular weight of 500,000 or more. including.
  • a method for alleviating knee joint pain in a human or non-human animal allows a human or non-human animal to orally ingest hyaluronic acid and / or a salt thereof having an average molecular weight of 500,000 or more. Including that.
  • the expression of SOCS3 is promoted by orally ingesting hyaluronic acid and / or a salt thereof having an average molecular weight of 500,000 or more, and symptoms of inflammatory diseases are reduced. Overall improvement is possible.
  • proiotrophin can be obtained by orally ingesting hyaluronic acid and / or a salt thereof having an average molecular weight of 500,000 or more. It becomes possible to comprehensively improve the symptoms of inflammatory diseases.
  • FIG. 1 is a photograph of electrophoresis in which SOCS3 gene expression by RT-PCR was examined in Example 1. From the left, the results of the distilled water intake group, the prototype A intake group, and the prototype B intake group are shown.
  • FIG. 2 is a photograph of the surface of the intestinal epithelium of a mouse orally ingested with hyaluronic acid or a salt thereof in Example 1.
  • FIG. 3 is a photograph of the surface of the large intestine epithelium of a mouse orally ingested with hyaluronic acid or a salt thereof in Example 1.
  • FIG. 1 is a photograph of electrophoresis in which SOCS3 gene expression by RT-PCR was examined in Example 1. From the left, the results of the distilled water intake group, the prototype A intake group, and the prototype B intake group are shown.
  • FIG. 2 is a photograph of the surface of the intestinal epithelium of a mouse orally ingested with hyaluronic acid or a salt
  • FIG. 4 is a photograph of the surface of the large intestine epithelium (after double staining treatment) of a mouse orally ingested with hyaluronic acid or a salt thereof in Example 1.
  • FIG. 5 is a photograph of a cervical lymph node of a mouse administered with drinking water of distilled water or a hyaluronic acid prototype in Example 1.
  • FIG. 6 is a photograph of electrophoresis in which SOCS3 gene expression was examined by RT-PCR in Example 2. From the left, the results are shown with no hyaluronic acid added, prototype A added, and prototype B added.
  • FIG. 5 is a photograph of a cervical lymph node of a mouse administered with drinking water of distilled water or a hyaluronic acid prototype in Example 1.
  • FIG. 6 is a photograph of electrophoresis in which SOCS3 gene expression was examined by RT-PCR in Example 2. From the left, the results are shown with no hyaluronic acid added, prototype A added, and
  • Example 7 shows that in Example 10, the knee joint space of STR mice orally fed hyaluronic acid of Test Example 4 and Test Example 5 was significantly higher than that of the control (distilled water) based on the soft radiographic image score. The results show that the narrowing is suppressed, and in Comparative Test Example 2 of chondroitin sulfate, there is no difference from the control.
  • FIG. 8 shows a soft X-ray image in Example 10. A white arrow points to the joint space. In FIG. 8, as compared with the narrow space of the control group (A), narrowing of the space of Test Example 4 (B) and Test Example 5 (C) in which hyaluronic acid was orally fed was suppressed. Comparative Test Example 2 (D) shows no difference from the control.
  • FIG. 8 shows that in Example 10, the knee joint space of STR mice orally fed hyaluronic acid of Test Example 4 and Test Example 5 was significantly higher than that of the control (distilled water) based on the soft radiographic image score. The results show that the narrow
  • FIG. 9 illustrates an evaluation method by Indian Ink method for knee joint cartilage of Example 10. This indicates that this is a method for evaluating the roughening of the cartilage surface.
  • FIG. 10 shows the roughening of the cartilage in Test Example 4 and Test Example 5 in which hyaluronic acid was orally fed according to the roughening index of cartilage in the knee joint cartilage of Example 10 evaluated by India Ink method. It shows the result that was significantly suppressed. Although the comparative test example 2 is suppressed somewhat, a significant difference is not recognized.
  • FIG. 11 shows a photograph of the surface of the knee joint cartilage stained with India Ink method in Example 10. In FIG.
  • FIG. 12 shows the results showing that the synovial activation and cell proliferation in Test Example 4 and Test Example 5 in which hyaluronic acid was fed orally were suppressed as compared with the control from the knee joint synovial evaluation score of Example 10. It is shown. The synovial membrane evaluation score of Comparative Test Example 2 is not different from the control.
  • SOCS3 expression promoter according to an embodiment of the present invention, a pharmaceutical and a food containing the same, a method for promoting the expression of SOCS3, a pleiotrophin expression inhibitor, a pharmaceutical and a food containing the same
  • a method for suppressing the expression of proiotrophin will be described in detail.
  • SOCS3 expression promoter pharmaceutical and food containing the same, and method for promoting SOCS3 expression
  • SOCS3 Expression Promoter The SOCS3 expression promoter according to one embodiment of the present invention is characterized by containing hyaluronic acid having an average molecular weight of 500,000 or more and / or a salt thereof as an active ingredient.
  • hyaluronic acid refers to a polysaccharide having one or more repeating structural units composed of disaccharides of glucuronic acid and N-acetylglucosamine.
  • the “hyaluronic acid salt” is not particularly limited, but is preferably a food or pharmaceutically acceptable salt, for example, sodium salt, potassium salt, calcium salt, zinc salt, magnesium salt, ammonium salt, etc. Is mentioned.
  • the average molecular weight of hyaluronic acid and / or a salt thereof used in the SOCS3 expression promoter according to this embodiment is 500,000 or more, preferably 600,000 or more, more preferably 500,000 to 1.6 million, and still more preferably 60 10,000 to 1.6 million.
  • the average molecular weight of hyaluronic acid and / or a salt thereof is less than 500,000, it is difficult to improve the symptoms of inflammatory diseases and the function of the immune system may be reduced.
  • the average molecular weight of hyaluronic acid and / or a salt thereof exceeds 1.6 million, it may be difficult to dissolve and the effect thereof may not be sufficiently exerted, or it may be difficult to add to foods.
  • the specific viscosity is measured by (Equation (1)), and the reduced viscosity at each concentration is calculated (Equation (2)).
  • a graph is drawn with the reduced viscosity on the vertical axis and the concentration (g / 100 mL) of the product converted to dry matter on the horizontal axis, and the intrinsic viscosity is determined from the intersection of the straight line connecting the points and the vertical axis. Substituting the intrinsic viscosity obtained here into Laurent's formula (formula (3)) to calculate the average molecular weight (TC Laurent, M. Ryan, A. Pietruszkiewicz,: BBA, 42, 476-485 (1960)) .
  • the hyaluronic acid contained in the SOCS3 expression promoter is basically a disaccharide unit in which the 1-position of ⁇ -D-glucuronic acid and the 3-position of ⁇ -DN-acetyl-glucosamine are combined. Having at least one disaccharide, and basically composed of ⁇ -D-glucuronic acid and ⁇ -DN-acetyl-glucosamine, wherein two or more disaccharide units are bonded, Alternatively, it may be a sugar to which these elements are bonded, and derivatives thereof such as those having a hydrolyzable protecting group such as an acyl group can also be used.
  • the sugar may be an unsaturated sugar, and examples of the unsaturated sugar include non-reducing terminal sugars, usually those having unsaturated carbon atoms between positions 4 and 5 of glucuronic acid.
  • Hyaluronic acid and / or a salt thereof may be extracted from natural products such as animals (for example, biological tissues such as chicken crown, umbilical cord, skin, joint fluid, etc.), or obtained by culturing microorganisms or animal cells. And the like (for example, fermentation using a bacterium belonging to the genus Streptococcus, etc.), or those synthesized chemically or enzymatically can be used.
  • animals for example, biological tissues such as chicken crown, umbilical cord, skin, joint fluid, etc.
  • microorganisms or animal cells obtained by culturing microorganisms or animal cells.
  • the like for example, fermentation using a bacterium belonging to the genus Streptococcus, etc.
  • those synthesized chemically or enzymatically can be used.
  • hyaluronic acid used in the SOCS3 expression promoter according to the present embodiment, a commercially available product can be used. For example, it can also be produced according to the following production methods 1 and 2.
  • Production method 1 extraction from chicken crown
  • the chicken crown is heat-treated. This is because the protein contained in the chicken crown is heat denatured or the enzyme is deactivated. Any method may be used for the heat treatment, but the heat treatment can be efficiently performed by immersing the chicken crown in hot water.
  • the heating temperature and time are not particularly limited as long as the protein in the chicken crown is denatured or the enzyme is deactivated.
  • a heating method using hot water a heat of 60 to 100 ° C. is used.
  • the raw material is preferably immersed in water for 20 to 90 minutes.
  • the chicken crown when a frozen chicken crown is used, the chicken crown may be heated as it is, but it is preferable to heat the frozen chicken crown after putting it in running water and then slowly thawing it so that a certain quality can be obtained.
  • Pasting improves the yield of hyaluronic acid.
  • An example of pasting is to add approximately 1 to 5 times the amount of fresh water to the chicken crown, and homogenize with a homogenizer for 10 to 60 minutes, so that the chicken crown is crushed and made into fine particles and can be finished into a paste.
  • acid agents such as hydrochloric acid and sulfuric acid, or alkali agents such as sodium hydroxide and potassium hydroxide are added to pasted chicken crowns to reduce the molecular weight of hyaluronic acid by acid treatment or alkali treatment.
  • the average molecular weight of the acid is adjusted to 500,000 or more.
  • the concentration of the acid agent or alkali agent, the amount added, and the treatment time may be appropriately combined so that the hyaluronic acid after treatment has a desired molecular weight. It is preferable because the molecular weight can be easily controlled.
  • An example of the alkali treatment is as follows.
  • an alkaline aqueous solution having a concentration of 10 to 30% is added to the chicken crown and treated at 25 to 70 ° C. for about 15 to 90 minutes. Neutralize with hydrochloric acid to adjust the molecular weight.
  • protease is added to the raw material whose molecular weight has been adjusted, and protease treatment is performed.
  • Any proteolytic enzyme can be used as long as it is commercially available, and examples thereof include pepsin, trypsin, papain, promeline and the like.
  • An appropriate amount of the protease is 0.01 to 1% with respect to the chicken crown.
  • the protease treatment temperature and time are preferably in the range of 35 to 65 ° C. and 1 to 10 hours.
  • hyaluronic acid is fractionated from the obtained protease-treated product to obtain crude hyaluronic acid, and then the hyaluronic acid is purified to obtain hyaluronic acid having a purity of 90% or more and an average molecular weight of 500,000 or more. It is done.
  • fractionation and purification of hyaluronic acid can be performed according to a conventional method.
  • the protease-treated raw material is filtered to remove solids to obtain a filtrate containing crude hyaluronic acid.
  • activated carbon may be added to the protease-treated product for the purpose of deodorization / decolorization or removal of some protein degradation products.
  • dissolving salt in the obtained filtrate ethanol is added and hyaluronic acid is precipitated, and a deposit is fractionated.
  • hydrous ethanol having an ethanol concentration of about 80 to 95% by volume is added to the precipitate, washed with a homogenizer, and the precipitate is collected.
  • the hyaluronic acid of Production Example 1 can be obtained by repeating this washing with hydrous ethanol about 2 to 10 times and drying the collected precipitate.
  • Production method 2 (microbe fermentation method) Hyaluronic acid calculation Activated carbon is added to the culture solution of Streptococcus microorganisms (Streptococcus zoopidemicus), followed by deodorization and decolorization, followed by filtration. Sodium chloride is dissolved in the obtained filtrate, ethanol is added to precipitate hyaluronic acid, and the precipitate is collected. Thereafter, hydrous ethanol having an ethanol concentration of about 80 to 95% by volume is added to the precipitate, washed with a homogenizer, and the precipitate is collected.
  • the hyaluronic acid (average molecular weight of 500,000 or more) of Production Example 2 can be obtained by repeating this washing with hydrous ethanol about 2 to 10 times and drying the collected precipitate.
  • the purity of hyaluronic acid used in the present invention may be a level that can be used in medicines or foods, preferably 90% or more, and more preferably 95% or more.
  • This purity is defined as a value obtained by removing impurities other than hyaluronic acid from 100% in terms of dry matter.
  • examples of the impurities include protein degradation products, fat (crude fat), chondroitin sulfate, and the like.
  • the purity of hyaluronic acid using chicken crown as a raw material can be obtained by the following formula (4).
  • Hyaluronic acid purity (%) 100-Proteolysate (%)-Crude fat (%)-Chondroitin sulfate (%)
  • the proteolysate (%) is a value obtained by the Lowry method
  • the crude fat (%) is a new food analysis method (published by Koso Co., Ltd.) “Chapter 1 General and related components, 1- 4 lipids, 1-4-2 ether extraction method ”, and chondroitin sulfate (%) is a value obtained by the method described below.
  • hyaluronic acid is dried, 50 mg of it is accurately weighed, and purified water is added to dissolve it.
  • Make exactly 100 mL to make a test solution take 4 mL of the test solution in a test tube, and add 1 mL of 0.5 mol / L sulfuric acid.
  • 0.2 mL of 0.04 mol / L cetyltrimethylammonium bromide was added to the solution obtained by heating in a water bath for 10 minutes and then cooling, and the mixture was allowed to stand at room temperature for 1 hour. Absorbance at a length of 10 mm and a wavelength of 660 nm is measured.
  • the obtained absorbance data is applied to a calibration curve for chondroitin sulfate to determine the amount (%) of chondroitin sulfate in purified hyaluronic acid.
  • the calibration curve was obtained by drying (depressurized, phosphorus pentoxide, 60 ° C., 5 hours) chondroitin sulfate A sodium salt (SG (Special Grade), manufactured by Seikagaku Corporation) derived from whale cartilage. Precisely weigh and dissolve in purified water, and prepare solutions containing 10 ⁇ g, 20 ⁇ g, 30 ⁇ g, and 40 ⁇ g of chondroitin sulfate A sodium salt in 1 mL, respectively.
  • SG Specific Grade
  • the promotion of SOCS3 expression in cells or tissues of human or non-human animals can be achieved by, for example, Northern blotting, detection or quantification of SOCS3 gene using a DNA array, DNA chip, etc., and Western blotting, ELISA, affinity chromatography, etc. It can be confirmed by a known biochemical analysis method such as detection or quantification of SOCS3.
  • the SOCS3 expression promoter according to one embodiment of the present invention usually contains 0.1% by mass or more of hyaluronic acid and / or a salt thereof having an average molecular weight of 500,000 or more, preferably 0.5 to 100% by mass. contains.
  • the average particle size of hyaluronic acid and / or a salt thereof is preferably 50 to 500 ⁇ m, more preferably 80 to 500 ⁇ m, and further preferably 200 to 500 ⁇ m.
  • the average particle size of hyaluronic acid and / or a salt thereof (hereinafter referred to as “hyaluronic acid” in this paragraph) having an average molecular weight of 500,000 or more (preferably 600,000 or more, more preferably 600,000 to 1.6 million) is 50 Since the powdered hyaluronic acid and / or salt thereof taken orally is dissolved in the gastric juice at an appropriate rate by being ⁇ 500 ⁇ m, the hydrolysis of hyaluronic acid in the gastric juice is suppressed, Hyaluronic acid can be absorbed from the intestinal tract while maintaining the molecular weight and can reach the affected area.
  • hyaluronic acid dissolves rapidly in the gastric juice and hydrolysis proceeds in the gastric juice, resulting in a decrease in molecular weight.
  • hyaluronic acid having a reduced molecular weight for example, a molecular weight of 500,000 or less
  • the average particle size of hyaluronic acid exceeds 500 ⁇ m, hyaluronic acid may not be completely dissolved in the body when ingested, and the amount absorbed may be less than the ingested amount. As a result, a sufficient effect for improving the symptoms of the inflammatory disease may not be obtained.
  • the average particle diameter of hyaluronic acid and / or a salt thereof is measured by a laser diffraction scattering method.
  • the average particle size of hyaluronic acid and / or a salt thereof can be adjusted by a commonly used method such as pulverization or sieving.
  • the pharmaceutical product according to one embodiment of the present invention contains the SOCS3 expression promoter according to the present embodiment as an active ingredient. More specifically, the pharmaceutical product according to the present embodiment can contain 5 to 100% by mass of the SOCS3 expression promoter according to the present embodiment.
  • the pharmaceutical product according to the present embodiment promotes the expression of SOCS3 in humans or non-human animals by orally ingesting human or non-human animals hyaluronic acid and / or a salt thereof having an average molecular weight of 500,000 or more. can do.
  • inflammatory diseases eg, rheumatoid arthritis (RA); asthma; allergic diseases such as rhinitis; vascular disease; thrombosis or harmful platelet aggregation; reocclusion after thrombolysis; reperfusion injury; psoriasis, eczema, Skin inflammatory diseases such as contact dermatitis and atopic dermatitis; diabetes (eg, insulin-dependent diabetes, autoimmune diabetes); multiple sclerosis; systemic lupus lupus erythematosus (SLE); ulcerative colitis, Crohn's disease Inflammatory bowel diseases such as (enteric enteritis) and cystitis (eg, bowel disease after colorectal resection and ileal anastomosis); childhood lipostool, non-tropical sprue, bowel disease associated with seronegative arthropathy, Diseases involving leukocyte infiltration into the gastrointestinal tract, such as lymphocytic or collagenous colitis, and
  • the foodstuff which concerns on one Embodiment of this invention contains the SOCS3 expression promoter which concerns on this embodiment. More specifically, the food according to the present embodiment can contain 0.1 to 100% by mass of the SOCS3 expression promoter according to the present embodiment.
  • the food may be intended to alleviate and improve symptoms of inflammatory diseases as a part of treatment or as an adjunct to treatment by the patient taking it over a long period of time. it can.
  • hyaluronic acid is observed on the intestinal epithelial surface of mice ingested by hyaluronic acid or its salt (See FIG. 2 and FIG. 3), the localization of TLR-4, which is a receptor on the intestinal epithelial surface, coincides with the localization of hyaluronic acid (see FIG. 4). It is speculated that hyaluronic acid and / or a salt thereof is due to promoting the expression of SOCS3.
  • the pharmaceutical product according to the present embodiment can contain other raw materials in addition to hyaluronic acid and / or a salt thereof, which is an SOCS3 expression promoter, as long as the effects of the present invention are not impaired.
  • raw materials are water, excipients, antioxidants, preservatives, wetting agents, thickeners, buffers, adsorbents, solvents, emulsifiers, stabilizers, surfactants, lubricants, Water-soluble polymers, sweeteners, flavoring agents, acidulants, alcohols and the like can be mentioned.
  • the dosage form of the pharmaceutical product according to this embodiment is not particularly limited, but when the pharmaceutical product is taken orally, for example, a solid preparation such as a tablet, powder, fine granule, granule, capsule, pill, etc., liquid, suspension Orally administrable agents such as liquids such as sachets, syrups and emulsions.
  • the food according to the present embodiment contains hyaluronic acid and / or a salt thereof, which is a SOCS3 expression agent, as an active ingredient.
  • hyaluronic acid and / or a salt thereof which is a SOCS3 expression agent
  • the aspect of the food is not particularly limited, for example, gum, candy, gummy candy, troche-like food, jelly beverage, cooked rice food, bread making, canned retort, frozen food, side dish, dried food, mayonnaise and other seasonings
  • General foods such as beverages, confectionery, desserts, and supplements, and general health foods that are permitted to express physiological functions are mentioned. Of these, supplements are preferable because of their convenience. .
  • the supplements may be in the form of solids such as tablets, powders, fine particles, granules, capsules (hard capsules, soft capsules), fluids, suspensions, jellies, syrups, etc. Is mentioned.
  • the said foodstuff can also be ingested by adding or mixing to the meal of an inflammatory disease.
  • any food and beverage can contain the food according to the present embodiment.
  • the SOCS3 expression promoter according to the present embodiment is continuously used. Therefore, it is desirable to include the SOCS3 expression promoter according to the present embodiment in foods and beverages that are ingested daily.
  • Hyaluronic acid and / or its salt is a biological substance, so even if it is ingested in large amounts, it is considered that there is no side effect or very low, but the amount of hyaluronic acid and / or its salt taken as a pharmaceutical is 10 mg per day
  • the standard can be -1000 mg, preferably 100-500 mg.
  • the number of administrations can be selected once or multiple times per day depending on the symptoms.
  • the amount of the hyaluronic acid and / or salt thereof of the present invention to be ingested as food can be 1 to 1000 mg, preferably 15 to 300 mg per day.
  • the SOCS3 expression promoter according to this embodiment is 0.001 to 1% by weight in the beverage, preferably 0.01 to 0.5. It can be contained by weight.
  • the proiotrophin expression inhibitor according to one embodiment of the present invention is the hyaluronic acid and / or its Contains salt as an active ingredient. That is, the hyaluronic acid and / or salt thereof contained in the pleiotrophin expression inhibitor according to this embodiment is hyaluronic acid and / or a salt thereof having an average molecular weight of 500,000 or more, and the above-mentioned SOCS3 expression promoter The same hyaluronic acid and / or a salt thereof exemplified in the column can be used.
  • pleiotrophin has been reported to be promoted by the development of autoimmune encephalomyelitis (Liu X, Mashour GA, Webster HF, Kurtz A, “Basic FGF and FGF receptor1 are expressed in microglia during experimenral autoimmune encephalomyelitis: temporally distinct expression of midkine and pleiotrophin ”, Glia., 24, 390-397, 1998.).
  • pleiotrophin expression is also promoted in inflammatory diseases such as rheumatism (PufePT, Bartscher M, Petersen W, Tillmann B, Mentlein R, “Expression of pleiotrophin, tropan embryonic growth and differentiation factor, in rheumatoid arthritis ”, Arthritis Rheum., 48, 660-667, 2003.).
  • the proiotrophin expression inhibitor according to one embodiment of the present invention contains hyaluronic acid having an average molecular weight of 500,000 or more and / or a salt thereof as an active ingredient, and thereby, for example, exhibits proiotrophin expression by oral ingestion. Can be suppressed.
  • the suppression of proiotrophin expression can be evaluated by, for example, the degree of expression of the proiotrophin gene by the DNA array, as shown in the Examples described later.
  • the method for suppressing the expression of pleiotrophin comprises orally ingesting hyaluronic acid and / or a salt thereof having an average molecular weight of 500,000 or more to a human or a non-human animal. Including.
  • Knee joint pain relieving agent As described above, the pharmaceutical product or food according to one embodiment of the present invention may be used to relieve knee joint pain.
  • hyaluronic acid and / or a salt thereof those described above can be used.
  • Degenerative knee joint disease initially causes pain at the beginning of the knee movement, such as when standing up, and when this progresses, pain appears during walking on the flat ground or during nighttime sleep, greatly restricting life and daily activities. It will be received.
  • ⁇ Degenerative knee joint diseases can be treated by taking anti-inflammatory analgesics or injecting sodium hyaluronate into the joints to reduce inflammation pain.
  • physical therapy such as muscle strengthening and thermotherapy is taken.
  • surgical procedures such as osteotomy and artificial joint replacement are taken. Both have a great mental and physical burden on the patient.
  • the knee joint pain relieving agent according to an embodiment of the present invention contains hyaluronic acid having an average molecular weight of 500,000 or more and / or a salt thereof as an active ingredient.
  • the knee joint pain may be due to a degenerative knee joint disease.
  • the method for alleviating knee joint pain according to one embodiment of the present invention includes orally ingesting hyaluronic acid having an average molecular weight of 500,000 or more or a salt thereof (for example, 60 to 300 mg per day).
  • hyaluronic acid having a specific molecular weight band and / or a salt thereof as an active ingredient, containing it as a medicine or food, and taking it in a predetermined amount orally, Can relieve knee pain and improve daily life and activities.
  • the usage form of the knee joint pain relieving agent according to the present embodiment is not particularly limited, and may be a usage form such as powder, granule, high-concentration liquid, and low-concentration liquid.
  • a usage form such as powder, granule, high-concentration liquid, and low-concentration liquid.
  • a dry form is preferable to a liquid form.
  • the compounding quantity of the hyaluronic acid of the knee joint pain relieving agent which concerns on this embodiment, or its salt can be suitably determined according to the usage form and the addition amount to a pharmaceutical or a foodstuff.
  • the knee joint pain relieving agent includes, as necessary, a bulking agent, a binder, a lubricant, a preservative, an antioxidant, a fragrance, a sweetener, a sour agent, an excipient, and the like. Can be blended.
  • vitamins such as vitamin C, vitamin B2, vitamin B12, and vitamin E, various nutritional components such as nutritional components such as nucleic acid, chondroitin sulfate, and collagen, and mineral components such as iron and zinc can also be blended.
  • the pharmaceutical or food containing the knee joint pain relieving agent according to one embodiment of the present invention can have the same composition as the pharmaceutical or food containing the above-described SOCS3 expression promoter.
  • the medicine or food according to one embodiment of the present invention may be used for treatment of the knee joint.
  • hyaluronic acid and / or a salt thereof those described above can be used.
  • Hyaluronic acid or a salt thereof is used as a therapeutic agent for knee joints, for example, republished patents WO00 / 53194, JP2002-348243, JP2004-181121, republished patent WO2005 / No. 040224, JP-A-11-60609, JP-A-2003-160464, and JP-A-10-43286.
  • these are techniques relating to so-called knee joint treatment injecting agents and artificial cartilage, in which a joint preparation containing hyaluronic acid or a salt thereof as an active ingredient is injected into the knee joint.
  • US Pat. No. 6,607,745 only describes the sensual effect of reducing knee joint pain due to ingestion.
  • Japanese Patent Application Laid-Open No. 2004-166616 describes the blood chondroitin sulfate concentration before and after administration and NTx (type 1 collagen cross-linked N-telopeptide), but it is a change in one biochemical parameter to the last, The effectiveness of knee osteoarthritis is not suggested.
  • Japanese Patent Publication No. 2003-530072 discloses that hyaluronic acid is considered to be effective for knee joint pain, and does not disclose clinical effects.
  • Japanese Patent Application Laid-Open No. 2007-314531 only describes the effect on chondrocytes from in vitro tests at the cellular level.
  • STR mice STR / OrtCrlj mice
  • the pathological anatomical observation of the joint revealed that the soft x-ray images showed a reduction in joint space reduction, cartilage roughening, and knee joint synovial degeneration, and knee joint disease, especially degenerative knee joint. It has been found that cartilage lesions due to diseases are suppressed, and the present invention has been completed.
  • the oral knee joint therapeutic agent according to one embodiment of the present invention contains hyaluronic acid and / or a salt thereof as an active ingredient.
  • the orally deformable knee joint disease therapeutic agent for oral use which concerns on one Embodiment of this invention uses hyaluronic acid and / or its salt as an active ingredient.
  • the oral knee joint cartilage therapeutic agent according to one embodiment of the present invention contains hyaluronic acid and / or a salt thereof as an active ingredient.
  • the therapeutic agent for oral knee joint synovium according to one embodiment of the present invention contains hyaluronic acid and / or a salt thereof as an active ingredient.
  • the method for alleviating knee joint pain according to one embodiment of the present invention includes orally ingesting hyaluronic acid having an average molecular weight of 500,000 or more or a salt thereof (for example, 60 to 300 mg per day).
  • hyaluronic acid and / or a salt thereof can be used as an active ingredient, and can be treated as a pharmaceutical or food, and can be treated orally ingested in a predetermined amount to treat knee joint diseases.
  • Daily life and activities can be improved by alleviating pain and removing obstacles to walking and movement.
  • the pharmaceutical or food containing the knee joint therapeutic agent according to one embodiment of the present invention can have the same composition as the pharmaceutical or food containing the SOCS3 expression promoter described above.
  • the average particle diameter of the powder is a value measured by dispersing the powder in ethanol and using SALD-2000A (manufactured by Shimadzu Corporation).
  • Example 1 In Example 1, the hyaluronic acid of the present invention was administered by drinking to MRL mice, and the SOCS3 gene expression by the SOCS3 expression promoter containing the hyaluronic acid and / or salt thereof of the present invention was investigated.
  • MRL mice are pathological model mice that spontaneously develop autoimmune diseases such as rheumatism, and various symptoms are known to occur due to autoimmune diseases.
  • mice Twelve MRL lpr / lpr (SPF) female mice (Charles River Japan, Inc.) 14 weeks old (weight approximately 27-37 g) were used in the experiment. Mice were raised in a breeding room set at 22 ⁇ 3 ° C. and a relative humidity of 50 ⁇ 20%, and a solid feed CE-2 for rat / mouse (CLEA Japan, Inc.) was fed as a feed.
  • hyaluronic acid prepared by the above-described production method hereinafter also referred to as “hyaluronic acid prototype” diluted with distilled water was used as a test substance, which was given freely. The details of the test substance are as follows.
  • Group 1 distilled water (Otsuka distilled water, manufactured by Otsuka Pharmaceutical Factory)
  • Group 2 Low molecular weight hyaluronic acid (prepared according to the above production method 1 to have an average molecular weight of 2000 (average molecular weight in the range of 400 to 20,000, average particle size: 67 ⁇ m), prototype A)
  • Group 3 hyaluronic acid (prepared according to the above production method 2 to have an average molecular weight of 900,000 (average molecular weight in the range of 600,000 to 1.6 million, average particle size: 75 ⁇ m), prototype B)
  • These test substances were prepared aseptically using a clean bench at room temperature. For the preparation, it was adjusted to a predetermined concentration using distilled water for injection and stored refrigerated until use.
  • the dose of hyaluronic acid prototype was set to 200 mg / kg / day, which is expected to be effective when administered with drinking water.
  • the amount of water consumed per day by the mouse was 6 mL / animal as a result of preliminary studies, and the body weight of the MRL mouse was about 33 g. Therefore, the concentration of prototype A and prototype B was set to 1.1 mg / mL.
  • pathological conditions such as lymphadenopathy spontaneously develop in MRL mice.
  • administration was started from 14 weeks of age, which is the initial stage of the disease state.
  • the dosing period was a 28-day study considered appropriate for evaluating the efficacy of drinking water.
  • mice After the 28-day intake test, the mice were euthanized and the small intestine, large intestine and cervical lymph nodes were removed. Using these samples, (1) confirmation of SOCS3 gene expression by DNA array, (2) confirmation of SOCS3 gene expression by RT-PCR, (3) localization of hyaluronic acid in the intestine, (4) cervix Lymph node weight was investigated.
  • AFFYMETRIX GeneChip Eukaryotic Target Preparation & Hybridization Manual
  • cDNA for RNA in a sample is synthesized, double-stranded cDNA is purified using GeneChip Sample Cleanup Module Kit (manufactured by AFFYMETRIX), and Ge neChip Expression 3'-Amplefication Reagents for IVT Labeling Kit ( AFFYMETRIX) was used to biotin-label the double-stranded cDNA to synthesize biotin-labeled cRNA, and the biotin-labeled cRNA was purified and fragmented using GeneChip Expression Sample Cleanup Module Kit (AFFYMETRIX).
  • hybridization control was added to this sample using GeneChip Eukaryotic Hybridization Control Kit (AF FYMETRIX), and hybridization was performed using GeneChip Hybridization Oven 640 (AFFYMETRIX) at 45 ° C for 16 hours at 60 rpm. . Subsequently, washing and streptavidin-phycoerythrin (molecular probe) staining were performed using GeneChip® Fluidic® Station® 450 (AFFYMETRIX). Furthermore, the DNA array obtained by using a laser scanner (GeneChip Scanner 3000-3G (AFFYMETRIX)) is scanned, and the obtained image is used with dedicated software (GeneChip Operating Software Ver. 1.4 (AFFYMETRIX))). And analyzed. As a result, SOCS3 gene enhancement was observed in the prototype B intake group.
  • the gene sequences of the probes fixed to the DNA array are shown in SEQ ID NOs: 1 to 11.
  • the probe information can be confirmed from the following website. https://www.affymetrix.com/site/login/login.affx (5 ⁇ ) TCACTTTTATAAAAATCCACCTCCA (3 ⁇ ) (SEQ ID NO: 1) (5 ⁇ ) GAGGCTGTCTGAAGATGCTTGAAAA (3 ⁇ ) (SEQ ID NO: 2) (5 ⁇ ) GAAGATGCTTGAAAAACTCAACCAA (3 ⁇ ) (SEQ ID NO: 3) (5 ⁇ ) GATGCTTGAAAAACTCAACCAAATC (3 ⁇ ) (SEQ ID NO: 4) (5 ⁇ ) TTGAAAAACTCAACCAAATCCCAGT (3 ⁇ ) (SEQ ID NO: 5) (5 ⁇ ) TCAACCAAATCCCAGTTCAACTCAG (3 ⁇ ) (SEQ ID NO: 6) (5 ⁇ ) CAACCAAATCCCAGTTCAACTCAGA (3 ⁇ ) (SEQ ID NO:
  • the SOCS3 gene expression was approximately doubled in the group administered with prototype B (hyaluronic acid having an average molecular weight of 900,000) as compared with mice that had been infused with distilled water.
  • prototype B hyaluronic acid having an average molecular weight of 900,000
  • SOCS3 gene expression was suppressed. Therefore, it was suggested that the expression of SOCS3 is promoted by oral ingestion of a hyaluronic acid prototype having an average molecular weight of 500,000 or more.
  • Relative / Optical / Intensity is an average value of Relative / Optical / Intensity for each probe of SEQ ID NOs: 12 and 13.
  • the SOCS3 gene expression was about 1.3 times higher in the group administered with prototype B (hyaluronic acid with an average molecular weight of 900,000) than in mice that had received distilled water. It was.
  • the group to which prototype A (hyaluronic acid having an average molecular weight of 2000) was administered had SOCS3 gene expression equivalent to that of the distilled water administration group. Therefore, it was suggested that the expression of SOCS3 is promoted by oral ingestion of a hyaluronic acid prototype having an average molecular weight of 500,000 or more.
  • the sample after the pretreatment was immersed in a 100 mM acetate buffer (pH 6.0) containing 200 TRU / mL hyaluronidase (derived from actinomycetes, Amano enzyme) at 60 ° C. for 120 minutes to fragment hyaluronic acid. This was washed with PBS for 5 minutes, and this washing step was repeated three times.
  • VECTOR Streptavidin-biotin Blocking Kit
  • hyaluronic acid is present on the epithelial surface of the small intestine and large intestine of the group administered with prototype B (hyaluronic acid having an average molecular weight of 900,000) (in FIG. 2 and FIG. 3).
  • the arrow indicates hyaluronic acid).
  • Table 4 shows the weight measurement results of the cervical lymph node
  • FIG. 1 shows a photograph of the cervical lymph node
  • FIG. 2 shows a photograph of the surface of the small intestine epithelium
  • FIG. 5 shows a photograph of the surface of the large intestine epithelium.
  • the weight of the cervical lymph node in the group administered with prototype A (hyaluronic acid having an average molecular weight of 2000) did not decrease significantly compared to the weight of the cervical lymph node in the group of mice drinking distilled water.
  • Example 2 In Example 2, HT29 cells, which are cell lines derived from human large intestine epithelium, were used, and the SOCS3 gene expression by the SOCS3 expression promoter containing hyaluronic acid and / or a salt thereof of the present invention was investigated.
  • HT29 cells were cultured under the following conditions.
  • Prototype A and prototype B used in Example 1 were used and added so that the hyaluronic acid concentration in the medium was 100 ng / mL. After culturing for 24 hours, it was subjected to RT-PCR in the same manner as in Example 1. In addition, the oligonucleotide which has a gene sequence shown by sequence number 14 and 15 was used as a primer, and the amplification cycle was 38 cycles.
  • 5 ⁇ ) GCCACCTACTGAACCCTCCT (3 ⁇ ) (SEQ ID NO: 14) 5 ⁇ ) ACGGTCTTCCGACAGAGATG (3 ⁇ ) (SEQ ID NO: 15)
  • Example 3 Using the prototype B of Example 1 (hyaluronic acid having an average molecular weight of 900,000) as an SOCS3 expression promoter as it was, soft capsules having the following composition were produced.
  • Example 4 Prototype B of Example 1 (hyaluronic acid having an average molecular weight of 900,000) was used as it was as an SOCS3 expression promoter to prepare a powder (granule) having the following composition.
  • Example 5 Prototype B of Example 1 (hyaluronic acid having an average molecular weight of 900,000) was used as it was as an SOCS3 expression promoter to produce tablets having the following composition.
  • Example 6 About the same sample as the sample evaluated in Example 1, pleiotrophin gene expression was confirmed by the same method as the confirmation of SOCS3 gene expression by the DNA array of Example 1.
  • the gene sequences of the probes fixed to the DNA array are shown in SEQ ID NOs: 16 to 26.
  • the probe information can be confirmed from the following website. https://www.affymetrix.com/site/login/login.affx (5 ⁇ ) AATGTATACCATAGTACCAGTAGGG (3 ⁇ ) (SEQ ID NO: 16) (5 ⁇ ) AGGAAGTTGAACTCTGTAGTACATA (3 ⁇ ) (SEQ ID NO: 17) (5 ′) GATTGAGGTAAGTTTTTTGGTGTTG (3 ′) (SEQ ID NO: 18) (5 ⁇ ) GTGATATTTCACATTTAAATCTTTT (3 ⁇ ) (SEQ ID NO: 19) (5 ⁇ ) ATGTTTTCTCTTGTGCATCAATTTA (3 ⁇ ) (SEQ ID NO: 20) (5 ⁇ ) GTGCATCAATTTAAATGTTACAACC (3 ⁇ ) (SEQ ID NO: 21) (5 ⁇ ) AACCATGTAAACTACTTCTCTTGTT (3 ⁇ ) (SEQ ID NO:
  • the fluorescence intensity (average value) by the DNA array is the average value of the fluorescence intensity for each probe of SEQ ID NOs: 5 to 16.
  • Example 7 By promoting the expression of SOCS3, it is expected that cytokine signaling is inhibited and the expression of inflammatory cytokines is suppressed. Therefore, the expression of inflammatory cytokines shown in Table 6 was investigated by performing cytokine array using the large intestine of mice ingesting hyaluronic acid according to the present invention (prototype B used in Example 1).
  • Example 1 and Example 2 it can be understood that the intake of prototype B promotes the expression of SOCS3 and can reduce inflammatory diseases. Moreover, according to the present Example, it turns out that the expression of IL-10 which is an anti-inflammatory cytokine was accelerated
  • prototype B hyaluronic acid
  • Example 8 When hyaluronic acid is dissolved in gastric juice at a low pH, the molecular weight of hyaluronic acid is reduced by hydrolysis, and the molecular weight of hyaluronic acid may be lower than the molecular weight of oral intake. For example, the improvement effect of knee arthritis may be reduced. For this reason, it is presumed that the lower the solubility of hyaluronic acid in water, the better the therapeutic effect of knee arthritis.
  • sieving is performed by sieving 1 (65 mesh (aperture 208 ⁇ m)), sieving 2 (150 mesh (aperture 104 ⁇ m)), sieving 3 (250 mesh (aperture 61 ⁇ m)) ) Sieve was used.
  • the particles that passed through the sieve 1 but did not pass through the sieve 2 were used as the particles of Test Example 1
  • the particles that passed through the sieve 2 but did not pass through the sieve 3 were used as the particles of Test Example 2, and did not pass through the sieve 1
  • These particles were used as particles of Test Example 3, and particles that passed through sieve 3 were used as particles of Comparative Test Example 1.
  • hyaluronic acid having an average molecular weight of 8,000 was sieved in the same manner to prepare hyaluronic acid having each average particle diameter shown in Table 8 (the preparation methods of the particles in Reference Examples 1 to 4 were respectively test examples). 1 to 3 and the preparation method of Comparative Test Example 1).
  • the hyaluronic acid of Test Examples 1 to 3 having an average particle size of 50 to 500 ⁇ m is less susceptible to hydrolysis in gastric juice than the hyaluronic acid of Comparative Test Example 1 having an average particle size of less than 50 ⁇ m, and has a low It can be seen that it is difficult to molecularize.
  • hyaluronic acid having an average particle size of 200 to 500 ⁇ m and an average molecular weight of 500,000 or more is not easily hydrolyzed by gastric juice and is not easily reduced in molecular weight. Therefore, it is absorbed from the intestinal tract while maintaining the molecular weight and reaches the affected area. Therefore, it is presumed that the therapeutic effect of inflammatory diseases is excellent.
  • Example 9 In Example 9, hyaluronic acid having an average molecular weight of 1 million (having an average molecular weight in the range of 600,000 to 1,200,000) and an average particle diameter of 205 ⁇ m was used as a detailed indication of subjective symptoms of knee joint pain, as shown in detail below. 15 males and females orally administer 240 mg a day in the form of soft capsules, and the course after ingestion, 4 weeks, and 8 weeks after ingestion, JKOM knee joint pain questionnaire and criteria for treatment results of knee osteoarthritis (JOA) It investigated according to.
  • JKOM knee joint pain questionnaire and criteria for treatment results of knee osteoarthritis (JOA) It investigated according to.
  • Hyaluronic acid having an average molecular weight of 1,000,000 was obtained according to the above production method 2, and this was sized to an average particle diameter of 205 ⁇ m, and then a soft capsule was prepared so that it would be 48 mg per grain. 5 soft capsules (240 mg as hyaluronic acid) were taken as daily intake.
  • JOA knee joint disease treatment outcome criteria
  • Kellgren-Lawrence osteoarthritis disease classification one of the methods for evaluating knee osteoarthritis X-rays Grades of 0 to 4 were classified according to the degree of symptom progression at the joint site), and grades of 1 to 3 were selected on either leg.
  • JKOM Knee Joint Pain Questionnaire 1 Evaluation was made based on the degree of knee pain (VAS: visual analog scale) and the change in the total value of criteria 2-5.
  • ⁇ Criteria 1 Knee pain level> When “no pain” is 1, and “most severe pain experienced so far” is 40, the degree of pain is tabulated with a score of 40 levels.
  • hyaluronic acid having an average molecular weight of 1,000,000 and an average particle size of 205 ⁇ m was orally administered to 15 men and women with subjective symptoms of knee joint pain in the form of soft capsules, 240 mg per day.
  • knee joint pain can be alleviated by orally ingesting a supplement containing hyaluronic acid having an average molecular weight of 500,000 or more and an average particle size of 50 to 500 ⁇ m as an active ingredient.
  • hyaluronic acid having an average molecular weight of 1,200,000 (with an average molecular weight in the range of 800,000 to 1,600,000) and an average particle size of 80 ⁇ m manufactured according to production method 2
  • the total score of JKOM knee joint pain assessment after 8 weeks was significantly improved with a risk rate of less than 1%.
  • the knee joint pain relieving agent of Example 8 is mainly composed of hyaluronic acid or a salt thereof having an average molecular weight of 500,000 or more (specifically, an average molecular weight of 600,000 to 1,600,000) and an average particle diameter of 50 to 500 ⁇ m.
  • the patient who complains of knee joint pain is orally ingested 120-240 mg per day for 8 weeks, so that the knee joint pain is alleviated.
  • JKOM knee joint pain evaluation and treatment results for degenerative knee joint disease are judged.
  • the total value of the reference (JOA) scores improved significantly with a risk rate of less than 1%. Therefore, this knee joint pain relieving agent and pharmaceuticals and foods containing the same are effective for alleviating knee joint pain.
  • Example 10 In Example 10, the two types of hyaluronic acid were each dissolved in drinking water and administered to STR mice as shown in detail below. Additive-free drinking water as a control and chondroitin sulfate similarly dissolved in drinking water were used as positive controls. After 17 weeks, both hind limbs of euthanized mice were used as autopsy samples. As autopsy items, (1) evaluation of narrowing of the space by soft X-ray images of the knee joint, (2) roughening evaluation of the knee joint cartilage surface, and (3) evaluation of degeneration of the knee joint synovium.
  • Test Product and Intake As Test Example 4, hyaluronic acid having an average molecular weight of 900,000 (those having an average molecular weight in the range of 600,000 to 1,600,000, average particle diameter: 159 ⁇ m) by extraction from a chicken crown according to Production Method 1. )
  • hyaluronic acid having an average molecular weight of 700,000 (those having an average molecular weight in the range of 500,000 to 1,200,000, average particle diameter: 211 ⁇ m) was obtained by a microbial fermentation method according to Production Method 2.
  • chondroitin sulfate (average molecular weight of about 30,000) was used. This was adjusted to 12.5 mg / mL in drinking water, and 7.5 mL per animal was ingested per day, so that the intake was 1000 mg / kg body weight / day (body weight approximately 40 g / animal).
  • Otsuka distilled water was prepared aseptically in a clean bench, and stored after refrigeration. This distilled water was used for the control group.
  • Test animals and test conditions STR / OrtCrlj mice male, 22 weeks old at the start of administration, produced by Charles River, Japan) were preliminarily raised for 15 days, and generally observed healthy mice with no abnormalities were subjected to the test.
  • Rearing was carried out in an environment of room temperature 19-25 ° C. and relative humidity 30-70% with 7 animals / cage using individual ventilation cage IVC.
  • the feed was reared by feeding the rat / mouse solid feed Quick Fat (manufactured by CLEA Japan, Inc.) and the drinking water by drinking the test product.
  • the evaluation was performed on the score evaluation of the joint space narrowing to evaluate whether or not the joint space partially or completely disappeared. At that time, compared with a normal joint image of a healthy mouse, 0 when there is no change, 0.5 when there is a slight change, 1 when there is a slight change, 2 when there is a moderate change, The case where there was a severe change was taken as 3.
  • Test Example 4 and Test Example 5 were significantly lower values than the control group, while Comparative Test Example 2 showed no difference from the control group.
  • the significance test was performed using the Kolmogorov-Smirnov (KS) test to determine whether there was a significant difference with a risk rate of less than 1%.
  • FIG. 8 shows soft X-ray images of 39-week-old STR mice in each group.
  • Test Example 4 (B) and Test Example 5 (C) have a wide joint space and suppress joint space reduction compared to the control group (A).
  • no difference is recognized in Comparative Test Example 2 (D).
  • the roughening evaluation of the knee joint cartilage surface was performed by India Ink Method. After applying Inia Ink to the surface of the femoral cartilage of the test sample of the hind limb fixed with formalin solution, the sample was washed in a physiological saline solution and photographed. India Ink remains in the roughened cartilage site. The cartilage area (number of pixels) was measured with a NIH image on a photograph as shown in FIG. Next, the number of grid intersections (excluding ligaments) where Indian Ink was stained was counted by Weigel's point counting method.
  • the value obtained by dividing the number by the cartilage area was defined as the cartilage roughening index.
  • Test Example 4 and Test Example 5 were significantly lower values than the control group, while Comparative Test Example 2 showed a slightly lower value than the control group, but no significant difference was observed. It was.
  • the significant difference test an average value and a standard error were calculated from the obtained data, a nonparametric or parametric Tukey multiple comparison test was performed, and the presence or absence of a significant difference was determined with a risk rate of less than 1%.
  • FIG. 10 shows a photograph of the surface of the femoral cartilage of 39-week-old STR mice in each group treated with India Ink.
  • Test Example 4 (B) and Test Example 5 (C) have a small stained area and the roughening of the knee joint cartilage surface is suppressed as compared with the control group (A).
  • Comparative Test Example 2 (D) had a slightly small stained area, but the difference was small.
  • Test Example 4 and Test Example 5 were lower than the control group, while Comparative Test Example 2 was not different.
  • Significant difference test calculated mean value and standard error from the obtained data, non-parametric or parametric Tukey multiple comparison test was performed, and significant difference was judged with less than 5% risk, but between any groups There was also no significant difference.
  • hyaluronic acid extracted from chicken crowns having an average molecular weight in the range of 600,000 to 1.6 million
  • hyaluronic acid obtained by microbial fermentation having an average molecular weight in the range of 500,000 to 1,200,000
  • a soft X-ray image of the knee joints of both hind limbs was obtained by orally feeding STR mice, which have a mean particle size in the range of 50-500 ⁇ m, over 17 weeks to STR mice, which are spontaneous knee joint disease models. From the images and histopathological examination results, suppression of narrowing of the knee joint space, suppression of roughening of the knee joint cartilage surface, and suppression of degeneration of the knee joint synovium were observed.
  • the knee joint therapeutic agent according to one embodiment of the present invention contains hyaluronic acid or a salt thereof as an active ingredient. Both radiographic image score and knee cartilage roughening index improved significantly with a risk rate of less than 1%. Although there was no significant difference in synovial scores on both sides of the patella, improvement was observed. Therefore, the knee joint therapeutic agent according to one embodiment of the present invention is effective in the treatment of degenerative knee joint disease, the treatment of knee joint cartilage, and the treatment of knee joint synovium. In addition, pharmaceuticals and foods containing these therapeutic agents are effective in improving the histopathological effects of the knee joint, and are effective in treating, preventing and alleviating diseases in the orthopedic field.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Health & Medical Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Veterinary Medicine (AREA)
  • Molecular Biology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Dermatology (AREA)
  • Epidemiology (AREA)
  • Biochemistry (AREA)
  • Materials Engineering (AREA)
  • Immunology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Rheumatology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Physical Education & Sports Medicine (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne un promoteur d'expression de SOCS3 qui contient de l'acide hyaluronique ayant un poids moléculaire moyen de 500 000 ou plus et/ou un sel de celui-ci en tant que composant actif.
PCT/JP2009/054491 2008-03-11 2009-03-10 Promoteur d'expression de socs3, médicament et aliment contenant celui-ci et procédé de promotion de l'expression de socs3 WO2009113512A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2010502813A JPWO2009113512A1 (ja) 2008-03-11 2009-03-10 Ｓｏｃｓ３発現促進剤、これを含有する医薬品および食品、ならびにｓｏｃｓ３の発現を促進する方法

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
JP2008060930 2008-03-11
JP2008-060930 2008-03-11
JP2008062032 2008-03-12
JP2008-062032 2008-03-12
JP2008263610 2008-10-10
JP2008-263610 2008-10-10

Publications (1)

Publication Number Publication Date
WO2009113512A1 true WO2009113512A1 (fr) 2009-09-17

Family

ID=41065179

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2009/054491 WO2009113512A1 (fr) 2008-03-11 2009-03-10 Promoteur d'expression de socs3, médicament et aliment contenant celui-ci et procédé de promotion de l'expression de socs3

Country Status (1)

Country Link
WO (1) WO2009113512A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013129577A1 (fr) * 2012-03-01 2013-09-06 キユーピー株式会社 Préparation orale pour stimuler l'expression de tgf-β, préparation orale pour inhiber la production de substance médiant la douleur, et préparation orale pour prévenir un œdème
JP2016518464A (ja) * 2013-03-14 2016-06-23 アナコティ リミテッド ヒアルロン酸誘導体
JP2017503819A (ja) * 2014-01-21 2017-02-02 ユタ−イナ ディーディーエス アンド アドバンスト セラピューティクス リサーチ センター 医療用導管用の生体内投与性マイクロ粒子
EP3685845A4 (fr) * 2017-09-19 2021-08-25 Laimu Corporation Composition et son procédé de production
JP7455990B2 (ja) 2021-11-30 2024-03-26 キユーピー株式会社 ヒアルロン酸粉末

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002223727A (ja) * 2001-01-31 2002-08-13 Ribateepu Seiyaku Kk 機能性食品
WO2002074318A1 (fr) * 2001-03-15 2002-09-26 Seikagaku Corporation Agents de regulation d'expression d'il-12
JP2005137334A (ja) * 2003-11-10 2005-06-02 Medicaraise Corp ヒアルロン酸含有ムコ多糖の製造方法
JP2007269806A (ja) * 1999-03-24 2007-10-18 Seikagaku Kogyo Co Ltd 人工唾液
JP2008266171A (ja) * 2007-04-18 2008-11-06 Q P Corp 自己免疫疾患緩和剤ならびにこれを含有する医薬品および食品

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007269806A (ja) * 1999-03-24 2007-10-18 Seikagaku Kogyo Co Ltd 人工唾液
JP2002223727A (ja) * 2001-01-31 2002-08-13 Ribateepu Seiyaku Kk 機能性食品
WO2002074318A1 (fr) * 2001-03-15 2002-09-26 Seikagaku Corporation Agents de regulation d'expression d'il-12
JP2005137334A (ja) * 2003-11-10 2005-06-02 Medicaraise Corp ヒアルロン酸含有ムコ多糖の製造方法
JP2008266171A (ja) * 2007-04-18 2008-11-06 Q P Corp 自己免疫疾患緩和剤ならびにこれを含有する医薬品および食品

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
TOSHIHIDE SATO ET AL.: "Keiko Hyalurone-san [Hiabesuto(J)] ni yoru Henkeisei Hiza Kansetsusho ni Taisuru Yukosei no Kento", SHIN'YAKU TO RINSHO, vol. 57, no. 2, 10 February 2008 (2008-02-10), pages 128 - 137 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013129577A1 (fr) * 2012-03-01 2013-09-06 キユーピー株式会社 Préparation orale pour stimuler l'expression de tgf-β, préparation orale pour inhiber la production de substance médiant la douleur, et préparation orale pour prévenir un œdème
JPWO2013129577A1 (ja) * 2012-03-01 2015-07-30 キユーピー株式会社 ＴＧＦ−β発現促進経口剤、痛み物質産生抑制経口剤及び浮腫抑制経口剤
JP2016518464A (ja) * 2013-03-14 2016-06-23 アナコティ リミテッド ヒアルロン酸誘導体
JP2017503819A (ja) * 2014-01-21 2017-02-02 ユタ−イナ ディーディーエス アンド アドバンスト セラピューティクス リサーチ センター 医療用導管用の生体内投与性マイクロ粒子
US10004690B2 (en) 2014-01-21 2018-06-26 Utah-Inha Dds & Advanced Therapeutics Research Center Administrable microparticles in vivo through medical conduit
EP3685845A4 (fr) * 2017-09-19 2021-08-25 Laimu Corporation Composition et son procédé de production
US11564950B2 (en) 2017-09-19 2023-01-31 Laimu Corporation Composition and method for producing the same
JP7455990B2 (ja) 2021-11-30 2024-03-26 キユーピー株式会社 ヒアルロン酸粉末

Similar Documents

Publication Publication Date Title
US20110053887A1 (en) Socs3 expression promoter, drug and food containing the same and method of promoting the expression of socs3
Boniel et al. Kabuki syndrome—clinical review with molecular aspects
CN102665736B (zh) 用于刺激肌肉蛋白合成的低热量高蛋白营养组合物
Blain et al. Crohn's disease clinical course and severity in obese patients
JP5554994B2 (ja) 乳酸菌含有製剤
EP2429533B1 (fr) Acides hyaluroniques sulfatés comme agents de régulation de l'activité cytokinique
WO2009113512A1 (fr) Promoteur d'expression de socs3, médicament et aliment contenant celui-ci et procédé de promotion de l'expression de socs3
Irwin et al. Intestinal inflammation without weight loss decreases bone density and growth
Wang et al. Secoisolariciresinol diglucoside suppresses Dextran sulfate sodium salt-induced colitis through inhibiting NLRP1 inflammasome
JP6218732B2 (ja) 変形性関節症予防又は治療用組成物
WO2020250980A1 (fr) Médicament oral, auxiliaire de suppression de tumeur et régime thérapeutique pour animal de compagnie
JPWO2011070767A1 (ja) アトピー性皮膚炎予防剤
JP5828018B2 (ja) リウマチ関節炎治療剤およびこれを含有する医薬品
JP2011042632A (ja) 経口用ドライアイ改善剤、ならびに前記経口用ドライアイ改善剤を含む食品組成物および医薬品組成物
JP5925569B2 (ja) 消化管ホルモン分泌調節剤
JP2017052747A (ja) ウロプラキン発現促進剤
CN101945665A (zh) 一种用于预防和治疗关节炎的含有蜘蛛酶的组合物
JP2003146888A (ja) 炎症性腸疾患予防治療剤
JP2010059084A (ja) 機能性食品及び医薬
JP2009102278A (ja) 膝関節痛緩和剤
JP7356710B2 (ja) ペリオスチン及びｐｋｍ２の分泌促進剤
JP2016193895A (ja) リーキーガットの予防又は改善用組成物
WO2024135235A1 (fr) Composition pour maintenir ou améliorer la fonction immunitaire
EP2252302B1 (fr) Composition pour le traitement de l'arthrose
JP2008266171A (ja) 自己免疫疾患緩和剤ならびにこれを含有する医薬品および食品

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 09720673

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2010502813

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 09720673

Country of ref document: EP

Kind code of ref document: A1