WO2018181071A1 - Composition for degrading non-collagenous glycoprotein - Google Patents

Composition for degrading non-collagenous glycoprotein Download PDF

Info

Publication number
WO2018181071A1
WO2018181071A1 PCT/JP2018/011923 JP2018011923W WO2018181071A1 WO 2018181071 A1 WO2018181071 A1 WO 2018181071A1 JP 2018011923 W JP2018011923 W JP 2018011923W WO 2018181071 A1 WO2018181071 A1 WO 2018181071A1
Authority
WO
WIPO (PCT)
Prior art keywords
bifidobacterium
bacterium
culture
composition
atcc
Prior art date
Application number
PCT/JP2018/011923
Other languages
French (fr)
Japanese (ja)
Inventor
琢磨 桜井
那波 橋倉
金忠 清水
Original Assignee
森永乳業株式会社
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 森永乳業株式会社 filed Critical 森永乳業株式会社
Priority to JP2019509744A priority Critical patent/JP6954995B2/en
Publication of WO2018181071A1 publication Critical patent/WO2018181071A1/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/195—Proteins from microorganisms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/745—Bifidobacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A61P27/06—Antiglaucoma agents or miotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/745—Blood coagulation or fibrinolysis factors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/46—Hydrolases (3)
    • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/46—Hydrolases (3)
    • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
    • A61K38/49—Urokinase; Tissue plasminogen activator

Definitions

  • the present invention relates to a non-collagenous glycoprotein degrading agent or a non-collagenous glycoprotein degrading composition, a non-collagenous glycoprotein degrading pharmaceutical composition and a non-collagenous glycoprotein degrading food / beverage composition.
  • Non-collagenous glycoprotein is a kind of cell adhesion molecule that forms a network structure of extracellular matrix surrounding cells and tissues.
  • Fibrinogen one of non-collagenous glycoproteins, is known to be involved in thrombus formation.
  • the blood coagulation system is activated along with the vasoconstriction, and fibrin is generated from fibrinogen, forming a fibrin clot and forming a thrombus.
  • Fibronectin and laminin are also known as non-collagenous glycoproteins that are components of the interface between the vitreous body of the eye and the retina.
  • the vitreous body of the eye is originally in close contact with the retina, but it is known that the vitreous body is denatured from a gel state to a liquid state with aging and peels from the retina (rear vitreous body peeling).
  • the posterior vitreous detachment itself is not a pathological change, but if the vitreous and the retina are firmly bonded, a retinal tear may be caused at the time of vitreal detachment and serious visual impairment may be caused.
  • Plasmin is classified as an endoprotease and exists in vivo as plasminogen, which is usually an inactive precursor.
  • a specific peptide bond of plasminogen is decomposed and activated by plasminogen activator (PA) to exert enzyme activity as plasmin.
  • PA plasminogen activator
  • Examples of plasminogen activators include tissue plasminogen activator (tPA) secreted from vascular endothelial cells and urokinase-type plasminogen activator (uPA) present in urine.
  • Plasmin is known to have an activity of degrading non-collagenous glycoproteins such as fibrinogen, fibronectin and laminin (Non-patent Document 1).
  • plasmin is an enzyme involved in a reaction that causes fibrinogenolysis and dissolves a fibrin clot, and has an action of dissolving a thrombus formed in the blood coagulation system. Therefore, plasmin is used in a conservative treatment method (fibrinolytic therapy) that prevents or reduces symptoms of ischemic injury caused by a thrombus formed by a fibrin clot (Non-patent Document 2).
  • Plasmin also has a degrading activity for fibronectin, which is a constituent component of the vitreous retinal interface, and is therefore used as an auxiliary substance when excising the vitreous from the retina (Patent Document 1).
  • the plasmin-containing fraction contains omega-amino acids.
  • Non-patent Document 3 an enzyme having plasmin activity derived from bacteria (natto) such as nattokinase enhances fibrinolytic activity in plasma.
  • natto contains abundant vitamin K produced by Bacillus natto, and by ingesting natto, vitamin K moves into the blood and reduces the efficacy of some antithrombotic drugs.
  • Non-Patent Document 4 and Non-Patent Document 5.
  • some Bifidobacterium bacteria that exist in the body bind to plasminogen on the cell surface and use the host plasminogen activator to bind to the cell surface. It has been reported that there is one having an activity of activating a gene and converting it into plasmin (Non-patent Document 6).
  • the Bifidobacterium genus, the culture of the bacterium, and / or the treated product of the bacterium have plasminogen activator activity and plasmin activity, thereby non-collagenous glycoprotein degradation effect It is not known to play.
  • the method for producing plasmin needs to undergo complicated steps so as not to impair plasmin activity. Moreover, when using natto kinase derived from natto, there is a concern about the side effect that vitamin K in natto promotes blood coagulation.
  • the present invention is a non-collagenous glycoprotein degrading agent or a non-collagenous glycoprotein degrading composition, a non-collagenous glycoprotein degrading pharmaceutical composition, and a non-collagenous glycoprotein degrading food and beverage that can be produced simply and without side effects.
  • An object is to provide a product composition.
  • the Bifidobacterium genus bacteria, the culture of the bacteria, and / or the treated product of the bacteria have plasminogen activator activity and plasmin activity. Based on this, it was found that a non-collagenous glycoprotein degradation effect was achieved, and the present invention was completed.
  • the present invention relates to a non-collagenous glycoprotein degrading agent or a composition for degrading non-collagenous glycoprotein, comprising as an active ingredient a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium. Offer things.
  • the degradation agent or the composition for degradation is preferably a mode in which the non-collagenous glycoprotein is fibrin, fibrinogen, fibronectin, laminin or plasminogen.
  • the decomposing agent or the decomposing composition is such that the Bifidobacterium genus bacterium is Bifidobacterium longum subspecies longum ATCC 15707 (Bifidobacterium longum subsp. Longum ATCC15707), Bifidobacterium longum subs. Spices Longum ATCC BAA-999 (Bifidobacterium longum subsp. Longum ATCC BAA-999) (or Bifidobacterium longum Subspecies longum BB536 (NITE BP-02621) (Bifidobacterium longum subsp. ))), Bifidobacterium longum subsp.
  • Infatis ATCC 15697 (Bifidobacterium longum subsp. Infantis tATCC15697), Bifidobacterium longum subsp. Infantis BCCM MG23728 (Bifidobacterium longum subsp. Infantis BCCM LMG23728) (or Bifidobacterium longum subspecies Infantis M-63 (NITE BP-02623) (Bifidobacterium longum subsp.
  • Bifidobacterium breve ATCC15700 Bifidobacterium breve ATCC15700
  • Bifidobacterium breve FERM BP-11175 Bifidobacterium breve BFERM BP-11175
  • Bifidobacterium breve BCCM LMG23729 Bifidobacterium breve GGCM Bifidobacterium breve M-16V (NITE BP-02622) (Bifidobacterium breve M-16V (NITE BP-02622))
  • Bifidobacterium bifidum ATCC29521 Bifidobacterium bifidum ATCC29521
  • Bi Fidobacterium adrecentis ATCC15703 Bifidobacterium adolescentis ATCC15703
  • Bifidobacterium angulatum ATCC 27535 Bifidobacterium breve ATCC15700
  • a preferred embodiment is one or more selected from the group consisting of:
  • the present invention provides a pharmaceutical composition for degrading non-collagenous glycoproteins comprising as an active ingredient a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium.
  • the pharmaceutical composition is cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction, cerebral venous sinus thrombus, retinal detachment, retinal tear, vitreous hemorrhage, diabetic vitreous hemorrhage, proliferative diabetes, age-related macular degeneration, macular circle It is preferably used for prevention and / or treatment of pores, vitreous macular traction, fibrin deposition, retinal vein occlusion, retinal artery occlusion, glaucoma or retinitis pigmentosa.
  • the present invention also provides a non-collagenized glycoprotein degrading food / beverage composition
  • a non-collagenized glycoprotein degrading food / beverage composition comprising as an active ingredient a Bifidobacterium genus bacterium, a culture of the bacterium, and / or a treated product of the bacterium.
  • the food / beverage product composition preferably contains plasminogen.
  • the present invention includes a step of culturing Bifidobacterium, and a step of recovering a protein having plasminogen activator activity and / or plasmin activity produced in the culture after the culture.
  • a method for producing a protein having plasminogen activator activity and / or plasmin activity is provided.
  • the present invention provides a method for producing plasmin, comprising a step of culturing Bifidobacterium in a medium containing plasminogen, and a step of recovering plasmin produced in the culture after the culture. provide.
  • the present invention provides use of a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium in the manufacture of a composition for degrading non-collagenous glycoprotein.
  • the present invention also provides a Bifidobacterium genus used for non-collagenous glycoprotein degradation, a culture of the bacterium, and / or a treated product of the bacterium.
  • the present invention also provides use of a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium for non-collagenous glycoprotein degradation.
  • the present invention also provides a method for degrading non-collagenous glycoprotein, comprising the step of administering a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium to a mammal. To do.
  • a non-collagenous glycoprotein degrading agent or a non-collagenous glycoprotein degrading composition, a non-collagenous glycoprotein degrading pharmaceutical composition, and a non-collagenous glycoprotein degrading agent that can be easily produced and have no side effects.
  • the food-drinks composition can be provided.
  • the non-collagenous glycoprotein degrading agent according to the present invention contains a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient.
  • the non-collagenous glycoprotein degrading agent according to the present invention contains a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient. It does not prevent the inclusion of ingredients. That is, the non-collagenous glycoprotein degrading agent according to the present invention is equivalent to the non-collagenous glycoprotein degrading composition.
  • another aspect of the present invention is a composition for degrading non-collagenous glycoproteins comprising a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient.
  • the content of Bifidobacterium in the composition is preferably 1 ⁇ 10 6 to 1 ⁇ 10 12 CFU / g or 1 ⁇ 10 6 to 1 ⁇ 10 12 CFU / mL, more preferably Is 1 ⁇ 10 7 to 1 ⁇ 10 11 CFU / g or 1 ⁇ 10 7 to 1 ⁇ 10 11 CFU / mL, more preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 CFU / g or 1 ⁇ 10 8 1 ⁇ 10 10 CFU / mL. If the bacterium is dead, the CFU can be replaced with cells.
  • the Bifidobacterium bacterium, the culture of the bacterium, and / or the treated product of the bacterium have plasminogen activator activity and plasmin activity, and based on these, non-collagenous glycoprotein This shows the decomposition effect. Therefore, in the present invention, plasminogen can be converted to plasmin even if plasminogen is not bound to the cell surface as in Non-Patent Document 6. That is, the present invention does not require a step of binding plasminogen to the cell surface of Bifidobacterium.
  • Another aspect of the present invention is a method for producing a composition for degrading non-collagenous glycoprotein, comprising a step of adding the Bifidobacterium of the present invention to a composition raw material.
  • the said manufacturing method contains the manufacturing method of the food-drinks composition including the process of adding the Bifidobacterium genus bacteria of this invention to the food-drinks composition raw material.
  • the said manufacturing method contains the manufacturing method of pharmaceutical composition including the process of adding Bifidobacterium genus bacteria to pharmaceutical composition raw materials (for example, base etc.).
  • the Bifidobacterium genus bacterium used in the method for producing the composition for degrading non-collagenous glycoprotein may be live or dead, and includes both live and dead But you can.
  • the Bifidobacterium bacterium may be added to the composition raw material after culturing, concentrating or freeze-drying, and the Bifidobacterium bacterium The Bifidobacterium may be cultured after adding to the composition raw material.
  • Bifidobacterium genus Bacteria belonging to the genus Bifidobacterium can be used in the genus Bifidobacterium, as long as the effects of the present invention are not impaired.
  • Infantis Bifidobacterium breve (Bifidobacterium breve), Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium angulatum, Bifidobacterium dentium, Bifidobacterium adolescentis, Bifidobacterium angulatum Bifidobacterium psudocatenulatum, Bifidobacterium animalis subsp. Lactis, Bifidobacterium anima Bifidobacterium animalis subsp. Animalis, Bifidobacterium pseudolongum subsp.
  • Bifidobacterium pseudolongum subsp. Globosum Bifidobacterium pseudolongum subsp. Pseudolongm
  • Bifidobacterium thermophilum etc.
  • the Bifidobacterium longum subspecies longum may be simply referred to as Bifidobacterium longum.
  • Bifidobacterium longum sub-species Infantis is sometimes simply referred to as Bifidobacterium Infantis.
  • Bifidobacterium longum subsp. Longum ATCC 15707 Bifidobacterium longum subsp. Longum ATCC15707
  • Bifidobacterium longum subspice longum ATCC BAA-999 Bifidobacterium longum subsp. Longlong ATCC BABA- 999
  • Bifidobacterium longum subspecies longis BB536 NITE BP-02621
  • Bifidobacterium longum subsp. Longum BB536 NITE BP-02621
  • Bifidobacterium longum subspecies Infatis ATCC 15697 Bifidobacterium longum subsp.
  • Bifidobacterium breve ATCC15700 Bifidobacterium breve ATCC15700
  • Bifidobacterium breve FERM BP-11175 Bifidobacterium breve FERM BP-11175
  • Bifidobacterium breve BCCM LMG23729 Bifidobacterium breve bc BCCM LMG23729
  • Bifidobacterium breve M-16V NITE BP-0222
  • Bifidobacterium breve M-16V Bifidobacterium breve M-16V (NITE BP-02622)
  • Bifidobacterium bifidum ATCC29521 Bifidobacteriumifbifidum ATCC29521
  • Bifidobacterium adrecentis ATCC15703 Bifidobacterium adol escentis ATCC15703
  • Bifidobacterium angulatum B
  • Bifidobacterium Pseudolongum subspecies pseudolongum ATCC25526 Bifidobacterium pseudolongum subsp. Pseudolongm ATCC25526
  • Bifidobacterium thermofilm ATCC25525 Bifidobacterium thermophilum ATCC25525
  • the Bifidobacterium only one kind may be used, or any two or more kinds may be used.
  • Bacteria with ATCC numbers can be obtained from the American Type Culture Collection (address: 12301 Parklawn Drive, Rockville, Maryland 20852, United States of America). Bacteria to which a BCCM number is assigned can be obtained from Belgian Coordinated Collections of Microorganisms (Address: B-1000 Brussels, Ciens Street (Wetens Cup Street) 8), Belgium.
  • Bacteria to which the FERM BP-11175 accession number has been assigned were issued on August 25, 2009 at the National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center (currently the National Institute for Product Evaluation Technology Patent Biological Deposit Center, 292-0818 2-5-8 Kazusa-Kamashita, Kisarazu City, Chiba Prefecture Room 120) has been deposited internationally based on the Budapest Treaty. Bacteria with DSM numbers can be obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (address: Inhoffenstra ⁇ e 7B, 38124 Braunschweig, Germany).
  • Bifidobacterium longum sub-species longum ATCC BAA-999 is the same bacterium as Bifidobacterium longum sub-species longum BB536 (NITE BP-02621). Bacteria may be used. Bifidobacterium longum subspecies longum BB536 (NITE BP-02621) was established on January 26, 2018, by the National Institute for Product Evaluation Technology Patent Microorganism Depositary Center (Kazusa, Kisarazu City, Chiba Prefecture 292-0818). International deposit based on the Budapest Treaty was made in Nama BP-02621 under No. 122 (Kamaashi 2-5-8 122).
  • Bifidobacterium longum subspecies Infatis BCCM LMG23728 is the same bacterium as Bifidobacterium longum subspecies Infantis M-63 (NITE BP-02623), which is a preferred embodiment. In, any bacteria may be used. Bifidobacterium longum subspecies Infantis M-63 (NITE BP-02623) was established on January 26, 2018, by the National Institute of Technology and Evaluation of the National Institute of Technology and Evaluation (JAPAN) In the prefecture, Kisarazu City, Kazusa Kamashi, 2-5-8 122), the deposit number of NITE BP-02623 was made based on the Budapest Treaty.
  • Bifidobacterium breve BCCM LMG23729 is the same bacterium as Bifidobacterium breve M-16V (NITE BP-02622), and any bacterium may be used in a preferred embodiment.
  • Bifidobacterium breve M-16V (NITE BP-02622) was issued on January 26, 2018, by the National Institute for Product Evaluation Technology Patent Microorganisms Depositary Center (Kazusa Kamashi 2, Kisarazu City, Chiba Prefecture 292-0818) -5-8 Room 122), the deposit number of NITE BP-02622, which was internationally deposited under the Budapest Treaty.
  • the Bifidobacterium genus bacterium of the present invention is not limited to the deposited bacterium, and may be a bacterium substantially equivalent to the deposited bacterium.
  • the bacterium substantially equivalent to the deposited bacterium is a bacterium belonging to the same genus or the same species as the deposited bacterium, and allows the subject to ingest the bacterium, a culture of the bacterium, and / or a treated product of the bacterium.
  • non-collagenous glycoprotein can be degraded in the subject, and the base sequence of the 16S rRNA gene is 98% or more, preferably 99% or more of the base sequence of the 16S rRNA gene of the deposited bacterium.
  • the bacterium has a homology of 100%, and preferably a bacterium having the same mycological properties as the deposited bacterium in addition to the homology.
  • the Bifidobacterium genus bacterium of the present invention is selected from the deposited bacterium or a bacterium substantially equivalent thereto, mutation treatment, genetic recombination, selection of a natural mutant, etc. Bacteria bred by may be used.
  • Bifidobacterium spp. Live in insects and small animals in addition to humans, but the habitat (host) is limited by the species, and Bifidobacterium spp.
  • the bacterium is called a human resident Bifidobacterium (HRB), otherwise it is called non-HRB.
  • HRB includes Bifidobacterium longum, Bifidobacterium longum subsp.
  • Bifidobacterium bacteria derived from infants are more preferable among HRBs. Examples of HRB derived from infants include Bifidobacterium longum, Bifidobacterium longum subsp. Infantis, Bifidobacterium breve, Examples thereof include Bifidobacterium bifidum.
  • the Bifidobacterium bacterium used in the present invention may be a mutant strain of the Bifidobacterium bacterium as long as it has a non-collagenous glycoprotein degrading effect equivalent to or better than the Bifidobacterium bacterium. . Whether or not a mutant strain has a “non-collagenous glycoprotein degradation effect equal to or higher than that of the above-mentioned Bifidobacterium genus” can be determined, for example, by measuring the plasmin activity of Bifidobacterium genus described later, It can be confirmed by measuring gen activator activity.
  • Such a mutant strain may be constructed by introducing a mutation artificially into the Bifidobacterium. In addition, it may be constructed by introducing mutation into the bacterium by treatment with a mutagen such as UV, or may be constructed by introducing mutation into the bacterium by various gene manipulation methods.
  • the plasmin activity is the fluorescence of a culture obtained by mixing a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium with a fluorescent substrate specific to plasmin, and culturing the mixture. This can be confirmed by measuring the strength. That is, the fluorescent substance concentration in the culture is calculated from the fluorescent intensity of the culture using a calibration curve indicating the relationship between the fluorescent substance concentration and the fluorescent intensity derived from the fluorescent substrate, and further, in one minute.
  • cells are separated from a culture of Bifidobacterium and suspended in a PBS solution to prepare a suspension, and the suspension is diluted to a turbidity (OD600) of 0.1.
  • Boc-Val-Leu-Lys-AMC manufactured by BACHEM
  • BACHEM Boc-Val-Leu-Lys-AMC
  • the enzyme activity (plasmin activity) of Bifidobacterium is determined from the fluorescence intensity of the culture measured at an excitation wavelength of 380 nm and a measurement wavelength of 460 nm with a fluorescence measuring device such as reader SH-9000 (Corona Electric). be able to.
  • a fluorescence measuring device such as reader SH-9000 (Corona Electric).
  • the plasmin activity is preferably 40 ⁇ U or more, more preferably 80 ⁇ U or more, and even more preferably 100 ⁇ U or more, the non-collagenous glycoprotein degradation effect of the present invention can be suitably exhibited.
  • plasminogen activator activity is not a plasmin-specific fluorescent substrate, but a plasminogen activator-specific fluorescent substrate as Z-Gly-Gly-Arg-AMC.HCl (BACHEM). It can be determined in the same manner as in the case of the plasmin activity except that the product is used. In the present invention, if the plasminogen activator activity is preferably 10 ⁇ U or more, more preferably 20 ⁇ U or more, and even more preferably 50 ⁇ U or more, the non-collagenous glycoprotein degradation effect of the present invention can be suitably exhibited. it can.
  • the Bifidobacterium bacterium used in the present invention and the culture of the bacterium can be easily obtained by culturing the Bifidobacterium bacterium by a conventional method.
  • the culture method is not particularly limited as long as Bifidobacterium can grow, and culture can be performed under appropriate conditions according to the nature of the bacteria.
  • the culture temperature may be 25 to 50 ° C., preferably 35 to 42 ° C.
  • cultivation on anaerobic conditions for example, it can culture
  • the medium for culturing Bifidobacterium used in the present invention is not particularly limited, and a medium usually used for culturing Bifidobacterium can be used. That is, as the carbon source, for example, saccharides such as glucose, galactose, lactose, arabinose, mannose, sucrose, starch, starch hydrolyzate, and molasses can be used depending on utilization. As the nitrogen source, for example, ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, and ammonium nitrate, and nitrates can be used.
  • the carbon source for example, saccharides such as glucose, galactose, lactose, arabinose, mannose, sucrose, starch, starch hydrolyzate, and molasses can be used depending on utilization.
  • the nitrogen source for example, ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, and am
  • inorganic salts examples include sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, and ferrous sulfate.
  • Organic components such as peptone, soybean powder, defatted soybean meal, meat extract, yeast extract and the like may also be used.
  • the Bifidobacterium genus bacterium used in the present invention can be used in the form of a bacterium itself, a culture of the bacterium, or a treated product of the bacterium as described above. That is, the bacterium belonging to the genus Bifidobacterium itself may be used, and after culturing the bacterium belonging to the genus Bifidobacterium, the obtained culture may be used as it is, or may be used after being diluted or concentrated. Moreover, the Bifidobacterium genus bacteria used for this invention may be live or dead, and may contain both live and dead.
  • the bacterial cell processed product examples include, for example, an immobilized bacterial cell in which the bacterial cell is fixed with acrylamide, carrageenan, or the like, and the cell wall and cell membrane of the bacterial cell are partially processed by an ordinary method such as ultrasonic treatment or homogenizer treatment. Or the completely crushed cell crushed material etc. are mentioned.
  • the cell disrupted product may be the whole fraction after the disruption or a part of the fraction, the centrifuged supernatant after the disruption, the fraction obtained by partially purifying the supernatant by ammonium sulfate treatment, or the above
  • the concentrate may be concentrated.
  • Bifidobacterium genus bacteria used in the present invention include, for example, a cell suspension thereof, a water-soluble fraction of the cell disruption product of the cells (bacteria), and a resuspension suspension.
  • the water-soluble fraction of the cell disruption product of the cells (bacteria) is preferable because of its high non-collagenous glycoprotein degradation activity.
  • the “bacterial cell suspension” in the present specification is a solution obtained by suspending a cell obtained by centrifuging a culture solution of the genus Bifidobacterium used in the present invention, preferably This is a solution obtained by the treatment described in (2) of Test Example 3 in Examples described later.
  • the “water-soluble fraction” in the present specification is a supernatant obtained by centrifuging the cell disrupted product, and is preferably described in (2) of Test Example 3 in Examples described later. It is a supernatant finally obtained by the process.
  • the water-soluble fraction is more preferably a water-soluble fraction appropriately purified or concentrated by a conventional method.
  • the “precipitation resuspension” in the present specification is a solution in which a precipitate obtained by centrifuging the above-mentioned cell disruption is suspended, and preferably in Test Example 3 of Examples described later. It is a solution in which a precipitate finally obtained by the treatment described in (2) is suspended.
  • Non-collagenous glycoprotein of the present invention includes cell adhesion molecules that form a network structure of an extracellular matrix surrounding cells and tissues, and may be in a monomer state or a polymer state.
  • fibrin also referred to as “stabilized fibrin”
  • fibrin polymer also referred to as “stabilized fibrin”
  • fibrinogen also referred to as “stabilized fibrin”
  • fibrin polymer fibrin monomer
  • fibrinogen also referred to as “stabilized fibrin”
  • fibrinogen also referred to as “stabilized fibrin”
  • fibrinogen also referred to as “stabilized fibrin”
  • fibrin polymer fibrin monomer
  • fibrinogen also referred to as “stabilized fibrin”
  • fibrinogen also referred to as “stabilized fibrin”
  • fibrinogen also referred to as “stabilized fibrin”
  • fibrinogen fibrinogen
  • fibronectin laminin
  • plasminogen vitronectin
  • nidogen tenascin
  • the non-collagenous glycoprotein degradation agent or non-collagenous glycoprotein degradation composition of the present invention can be used as a pharmaceutical composition.
  • Plasmin is a retinal detachment, retinal tear, vitreous hemorrhage, diabetic vitreous hemorrhage, proliferative diabetes, age-related macular degeneration, macular hole, vitreous macular traction, fibrin deposition, retinal vein occlusion, retinal artery occlusion, Chemical vitrectomy that requires vitreal detachment for glaucoma and retinitis pigmentosa; fibrinolysis for ischemic disorders such as cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction and cerebral venous sinus thrombus It is used for therapy (Patent Document 2, Non-Patent Document 1).
  • plasmin has a fibrinogen decomposing effect because it is effective for fibrinolytic therapy for ischemic disorders such as cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction and cerebral venous sinus thrombus (non- Patent Document 1).
  • the pharmaceutical composition of the present invention comprises retinal detachment, retinal laceration, vitreous hemorrhage, diabetic vitreous hemorrhage, proliferative diabetes, age-related macular degeneration, macular hole, vitreous macular traction, fibrin deposition, retinal vein occlusion Prevention and / or treatment of ischemic disorders such as cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction and cerebral venous sinus thrombosis Can be used for Moreover, since the pharmaceutical composition of the present invention contains a Bifidobacterium genus bacterium, a culture of the bacterium, and / or a treated product of the bacterium that have been used for many years as an oral composition component, It can be administered with peace of mind to patients suffering from various diseases.
  • the non-collagenous glycoprotein degrading agent or the non-collagenous glycoprotein degrading composition of the present invention comprises a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient. Therefore, it can be safely administered to infants and children. Therefore, the non-collagenous glycoprotein degrading agent or the non-collagenous glycoprotein degrading composition of the present invention is suitable for the prevention and / or treatment of diseases of infants and children.
  • the pharmaceutical composition may be administered either orally or parenterally. Accordingly, it can be appropriately formulated into a desired dosage form.
  • a desired dosage form for example, in the case of oral administration, it can be formulated into solid preparations such as powders, granules, tablets and capsules; liquid preparations such as solutions, syrups, suspensions and emulsions.
  • parenteral administration it can be formulated into a suppository, an ointment, an eye drop or the like.
  • non-collagenous glycoprotein degrading agent or non-collagenous glycoprotein degrading composition according to the present invention excipients, pH adjusters, and colorants that are usually used for formulation. Ingredients such as flavoring agents can be used.
  • the non-collagenous glycoprotein degrading agent or the non-collagenous glycoprotein degrading composition according to the present invention and known or future non-collagenous glycoproteins are involved.
  • a component having an effect of preventing and / or treating a disease can be used in combination.
  • formulation can be performed by a known method as appropriate according to the dosage form.
  • a formulation carrier may be appropriately blended to formulate.
  • the intake or dose of the pharmaceutical composition of the present invention can be appropriately selected according to the dosage form.
  • the daily intake or dosage of Bifidobacterium per kg body weight is preferably 1 ⁇ 10 6 to 1 ⁇ 10 12 CFU / kg / day, and 1 ⁇ 10 7 to 1 ⁇ 10 11 CFU. / Kg / day is more preferable, and 1 ⁇ 10 8 to 1 ⁇ 10 10 CFU / kg / day is more preferable.
  • CFU is an abbreviation of colony forming units and is a colony forming unit. If the bacterium is dead, the CFU can be replaced with cells.
  • the intake or dose is the above when converted to the intake or dose of Bifidobacterium. It is preferable to be an intake or a dose.
  • the content of Bifidobacterium in the pharmaceutical composition of the present invention can be appropriately selected based on the intake or dose.
  • 1 ⁇ 10 6 to 1 ⁇ 10 12 CFU / g or 1 ⁇ 10 6 to 1 ⁇ 10 12 CFU / mL preferably 1 ⁇ 10 7 to 1 ⁇ 10 11 CFU / g or 1 ⁇ 10 7 to 1 ⁇
  • It can be 10 11 CFU / mL, more preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 CFU / g or 1 ⁇ 10 8 to 1 ⁇ 10 10 CFU / mL.
  • the CFU can be replaced with cells.
  • the content may be the above content when converted to the content of Bifidobacterium bacteria. preferable.
  • preparation carrier various organic or inorganic carriers can be used depending on the dosage form.
  • examples of the carrier in the case of a solid preparation include excipients, binders, disintegrants, lubricants, stabilizers, and flavoring agents.
  • excipient examples include sugar derivatives such as lactose, sucrose, glucose, mannitol and sorbit; starch derivatives such as corn starch, potato starch, ⁇ -starch, dextrin and carboxymethyl starch; crystalline cellulose, hydroxypropyl cellulose, Cellulose derivatives such as hydroxypropylmethylcellulose, carboxymethylcellulose, carboxymethylcellulose calcium; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, magnesium magnesium magnesium silicate; phosphate derivatives such as calcium phosphate; And carbonate derivatives such as calcium; sulfate derivatives such as calcium sulfate and the like.
  • sugar derivatives such as lactose, sucrose, glucose, mannitol and sorbit
  • starch derivatives such as corn starch, potato starch, ⁇ -starch, dextrin and carboxymethyl starch
  • crystalline cellulose hydroxypropyl cellulose
  • binder examples include gelatin, polyvinyl pyrrolidone, macrogol and the like in addition to the above excipients.
  • disintegrant examples include, in addition to the above excipients, chemically modified starch or cellulose derivatives such as croscarmellose sodium, sodium carboxymethyl starch, and crosslinked polyvinylpyrrolidone.
  • talc stearic acid
  • stearic acid metal salts such as calcium stearate and magnesium stearate
  • colloidal silica waxes such as pea gum and geirow
  • boric acid glycol
  • carboxylic acids such as fumaric acid and adipic acid
  • Carboxylic acid sodium salts such as sodium benzoate
  • sulfates such as sodium sulfate; leucine
  • lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate
  • silicic acids such as anhydrous silicic acid and silicic acid hydrate; starch derivatives and the like It is done.
  • the stabilizer examples include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; sorbic acid and the like.
  • flavoring agent examples include sweeteners, acidulants, and fragrances.
  • a carrier used in the case of a liquid for oral administration a solvent such as water, a flavoring agent and the like can be mentioned.
  • the non-collagenous glycoprotein degrading agent or the non-collagenous glycoprotein degrading composition of the present invention can be used as a food or drink composition.
  • the food / beverage composition of the present invention may be produced by adding a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium to a known food / beverage product. It is also possible to produce a new food / beverage product composition by mixing the bacteria belonging to the genus Fidobacterium, the culture of the bacteria, and / or the processed product of the bacteria into the raw material of the food / beverage product.
  • the food-drinks composition of this invention can also be manufactured by the manufacturing method including the process of culture
  • plasmin is also used for ripening dairy products and producing cheese (X. G. Song et al., Journal of the Japan Food Industry Association, Vol. 40, No. 4, April 1993).
  • plasmin exists as plasminogen in mammalian milk, and it is known that the mother's milk of a premature mother has a higher plasmin concentration than usual.
  • the non-collagenous glycoprotein degradation food / beverage composition of the present invention preferably contains plasminogen, an additive food material for promoting ripening of dairy products and cheese, and a preparation for childcare for premature infants. It can be suitably used as a food material for addition to powdered milk, breast milk or food.
  • the food / beverage composition in the present invention may be in the form of liquid, paste, solid, powder, etc.
  • liquid food, feed including for pets
  • the food / beverage composition of the present invention may use a component having a probiotic effect known in the future or a component assisting the probiotic effect. it can.
  • the food / beverage composition of the present invention comprises various proteins such as whey protein, casein protein, soy protein, pea protein (pea protein) or mixtures thereof, and degradation products thereof; amino acids such as leucine, valine, isoleucine or glutamine; Vitamins such as vitamin B6 or vitamin C; creatine; citric acid; fish oil; or components such as isomalt-oligosaccharide, galactooligosaccharide, xylo-oligosaccharide, soybean oligosaccharide, fructooligosaccharide, lactulose .
  • proteins such as whey protein, casein protein, soy protein, pea protein (pea protein) or mixtures thereof, and degradation products thereof; amino acids such as leucine, valine, isoleucine or glutamine; Vitamins such as vitamin B6 or vitamin C; creatine; citric acid; fish oil; or components such as isomalt-oligosaccharide, galactooligosaccharide,
  • the food / beverage product composition defined in the present invention is used for prevention of diseases in which non-collagenous glycoproteins effectively act, risk reduction of diseases, symptom alleviation of diseases, and / or treatment (including health uses).
  • the “display” act includes all acts for informing the consumer of the use, and if the expression can remind the user of the use, the purpose of the display, the content of the display, the display Regardless of the target object / medium, etc., all fall under the “display” act of the present invention.
  • the “display” is performed by an expression that allows the consumer to directly recognize the use. Specifically, it is the act of transferring, displaying, importing, displaying, or importing products that are related to food or drinks or products that describe the use, on advertisements, price lists, or transaction documents. For example, an act of describing and displaying the above uses or distributing them, or describing the above uses in information including the contents and providing them by an electromagnetic (Internet or the like) method can be given.
  • the display content is preferably a display approved by the government or the like (for example, a display that is approved based on various systems determined by the government and is performed in a mode based on such approval).
  • labeling includes health food, functional food, enteral nutrition food, special purpose food, health functional food, food for specified health use, nutrition functional food, functional label food, quasi-drug, etc.
  • a display is also included.
  • indications approved by the Consumer Affairs Agency for example, indications approved in systems related to foods for specified health use, functional nutritional foods, functional indication foods, or similar systems, etc. can be mentioned.
  • labeling as a food for specified health use labeling as a conditionally specified food for specified health use, labeling that affects the structure and function of the body, labeling for reducing the risk of disease, and functionality based on scientific evidence Labeling, etc., and more specifically, Cabinet Office Ordinance concerning permission for special purpose labeling provided for in the Health Promotion Act (Cabinet Office Ordinance No. 57, August 31, 2000)
  • the labeling as food for specified health (particularly the labeling of health use) and the like are the typical examples.
  • the intake of the food / beverage product composition of the present invention can be appropriately selected.
  • the daily intake of Bifidobacterium per kg of body weight is preferably 1 ⁇ 10 6 to 1 ⁇ 10 12 CFU / kg / day, and 1 ⁇ 10 7 to 1 ⁇ 10 11 CFU / kg / day. Day is more preferable, and 1 ⁇ 10 8 to 1 ⁇ 10 10 CFU / kg / day is more preferable. If the bacterium is dead, the CFU can be replaced with cells.
  • the intake may be the above intake when converted to the intake of Bifidobacterium. preferable.
  • the content of Bifidobacterium in the food and beverage composition of the present invention can be appropriately selected based on the intake.
  • 1 ⁇ 10 6 to 1 ⁇ 10 12 CFU / g or 1 ⁇ 10 6 to 1 ⁇ 10 12 CFU / mL preferably 1 ⁇ 10 7 to 1 ⁇ 10 11 CFU / g or 1 ⁇ 10 7 to 1 ⁇
  • It can be 10 11 CFU / mL, more preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 CFU / g or 1 ⁇ 10 8 to 1 ⁇ 10 10 CFU / mL.
  • the CFU can be replaced with cells.
  • the content may be the above content when converted to the content of Bifidobacterium bacteria. preferable.
  • the non-collagenous glycoprotein degrading food / beverage composition according to the present invention can be used as a human or animal food / beverage composition.
  • the non-collagenous glycoprotein degradation food / beverage composition according to the present invention includes cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction, cerebral venous sinus thrombus, retinal detachment, retinal tear, vitreous hemorrhage, diabetic vitreous Non-collagenous glycoprotein-related diseases such as bleeding, proliferative diabetes, age-related macular degeneration, macular hole, vitreous macular traction, fibrin deposition, retinal vein occlusion, retinal artery occlusion, glaucoma or retinitis pigmentosa It is effective for prevention and / or treatment.
  • the food and drink composition for degrading non-collagenous glycoprotein of the present invention contains Bifidobacterium, culture of the bacterium, and / or treated product of the bacterium as an active ingredient. Can also be safely administered. Therefore, the non-collagenous glycoprotein degradation food / beverage composition of the present invention is also suitable for the prevention and / or treatment of diseases in infants and children.
  • Another embodiment of the present invention is a method for producing a protein having plasminogen activator activity and / or plasmin activity.
  • a step of culturing Bifidobacterium (culture step), and a step of recovering a protein having plasminogen activator activity and / or plasmin activity produced in the culture after the culture (Recovery step) is included, but other steps may be included.
  • the protein having plasminogen activator activity and the protein having plasmin activity may be one protein or different proteins. That is, one protein may have both plasminogen activator activity and plasmin activity, or a protein having plasminogen activator activity and a protein having plasmin activity may be separate proteins. .
  • This step is a step of culturing Bifidobacterium bacteria, and known culture conditions that can culture Bifidobacterium bacteria can be employed. For example, it can be cultured at 25 to 45 ° C., but it is preferably cultured at 30 to 42 ° C., more preferably 37 to 42 ° C. Moreover, it is preferable to perform culture
  • This step is a step of recovering a protein having plasminogen activator activity and / or plasmin activity produced in the culture after the culture.
  • This step includes (a) separating the culture after the culture into a Bifidobacterium genus and a fraction containing a protein having plasminogen activator activity and / or plasmin activity, and (b)
  • the method includes a step of recovering a protein having plasminogen activator activity and / or plasmin activity from the fraction, but may include other steps. Steps (a) and (b) may be performed simultaneously, and step (b) may be performed after step (a).
  • a known method can be employed, and for example, a method such as filtration with a membrane or centrifugation can be employed.
  • the membrane may be either a flat membrane or a hollow fiber membrane (hollow fiber).
  • the step (a) may include a step of treating cells of the genus Bifidobacterium. That is, the step (a) is separated into a step of treating cells of the genus Bifidobacterium and a fraction containing the treated product and a protein having plasminogen activator activity and / or plasmin activity. And a step of performing. About the process of processing the microbial cell of Bifidobacterium genus, the description regarding the microbial cell processed material already demonstrated is used.
  • step (b) a known method can be employed, and examples thereof include various chromatography methods such as gel filtration chromatography and reverse phase HPLC. Chromatography may be low or high pressure. When filtration is performed using a hollow fiber membrane (hollow fiber), the steps (a) and (b) can be performed simultaneously.
  • the fraction containing a protein having plasminogen activator activity and / or plasmin activity contains other components such as medium components as long as the effect of the protein having plasminogen activator activity and / or plasmin activity is exhibited.
  • the component or the like may be completely or partially purified, and the mode is not particularly limited.
  • purification can be performed by combining suitably the method in the said process (a) and / or (b).
  • the properties of the fraction containing a protein having plasminogen activator activity and / or plasmin activity may be liquid or powder obtained by lyophilization or the like, and the mode is not particularly limited.
  • the protein having plasminogen activator activity and / or plasmin activity produced according to the present embodiment is based on the physiological action of the protein, quasi-drug, external preparation for skin, cosmetics, food and drink, food. It can mix
  • Another embodiment of the present invention is a method for producing plasmin.
  • This embodiment includes a step of culturing Bifidobacterium in a medium containing plasminogen (culturing step), and a step of recovering plasmin produced in the culture after the culturing (recovering step).
  • culturing step a step of culturing Bifidobacterium in a medium containing plasminogen
  • recovering step recovering plasmin produced in the culture after the culturing
  • This step is a step of culturing Bifidobacterium in a medium containing plasminogen.
  • plasminogen When plasminogen is added to the medium, it may be added before culturing or during culturing. Moreover, any of collective addition, sequential addition, and continuous addition may be sufficient.
  • the content of plasminogen in the medium is not particularly limited.
  • the description of the “culturing step” in ⁇ Method for producing protein having plasminogen activator activity and / or plasmin activity> is incorporated in the culture conditions of Bifidobacterium.
  • This step is a step of recovering plasmin produced in the culture after the culture.
  • This step includes (c) a step of separating the culture after the culture into a fraction containing Bifidobacterium and plasmin, and (d) a step of recovering plasmin from the fraction. These steps may be included. Steps (c) and (d) may be performed simultaneously, and step (d) may be performed after step (c). Further, the step (c) may include a step of treating cells of the genus Bifidobacterium. That is, the step (c) may include a step of treating the cells of the genus Bifidobacterium, and a step of separating the treated cells and a fraction containing plasmin.
  • the plasmin produced according to this embodiment can be blended in compositions such as pharmaceuticals, quasi drugs, external preparations for skin, cosmetics, foods and drinks, food additives, and feeds based on the physiological functions of the plasmin. Moreover, the plasmin produced by this embodiment can be suitably used as a food material for addition to dairy products and cheese, a formula powder for childcare for premature infants, breast milk or a food material for addition to food.
  • the present invention can employ the following configurations.
  • the following mammals include humans, cows, sheep, goats, pigs, dogs, cats, horses and the like.
  • Bifidobacterium genus bacteria used for the alleviation, prevention or treatment of diseases that can be alleviated, prevented or treated by degradation of non-collagenous glycoprotein, culture of the bacteria, and / or cell treatment of the bacteria object.
  • a method for degrading non-collagenous glycoprotein comprising administering a Bifidobacterium genus bacterium, a culture of the bacterium, and / or a treated product of the bacterium to a mammal.
  • a non-collagenous glycoprotein degrading agent or a composition for degrading non-collagenous glycoprotein comprising a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient
  • a method for degrading non-collagenous glycoproteins comprising the step of administering to an animal.
  • It is alleviated, prevented or treated by degradation of non-collagenous glycoprotein, comprising administering a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium to a mammal.
  • a method for alleviating, preventing or treating a disease obtained.
  • a non-collagenous glycoprotein degrading agent or a composition for degrading non-collagenous glycoprotein comprising a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient
  • a method for alleviating, preventing or treating a disease that can be alleviated, prevented or treated by degradation of non-collagenous glycoprotein, comprising a step of administering to an animal.
  • Test Example 1 A test was conducted to confirm that Bifidobacterium has plasminogen activator activity.
  • the MRS liquid medium is prepared by dissolving 5.5 g of Difco Lactobacilli MRS Broth (BD) and 50 mg of L-Cysteine Monohydrochloride, Monohydrate (Wako Pure Chemical Industries) in pure water so as to be 100 mL, and using an aqueous HCl solution. It was prepared by adjusting to pH 6.5 and sterilizing at 121 ° C. for 15 minutes.
  • HRB Human resident Bifidobacterium
  • the fluorescence intensity of the culture was measured using a microplate reader SH-9000 (manufactured by Corona Electric) at an excitation wavelength of 360 nm and a measurement wavelength of 460 nm.
  • the unit of fluorescence intensity was an arbitrary unit (au).
  • the AMC concentration in the culture is calculated using a calibration curve showing the relationship between the fluorescent substance-derived fluorescent substance (AMC: aminomethylcoumarin) concentration prepared in advance and the fluorescence intensity at a measurement wavelength of 460 nm. From this concentration, the enzyme activity (plasminogen activator activity) of each Bifidobacterium was calculated using 1 nmol of AMC produced per minute as 1 unit of enzyme (U).
  • Bifidobacterium bacterium and / or the Bifidobacterium culture according to the present invention exhibited plasminogen activator activity and / or plasmin activity. It was suggested to have a glycoprotein degradation effect.
  • Test Example 1 One or more selected from 17 kinds of Bifidobacterium used in Test Example 1 was added to 3 mL of each or the same MRS liquid medium, anaerobically cultured at 37 ° C. for 16 hours, and the culture solution was concentrated. Freeze-drying is performed to obtain bacterial powder of the one or more bacteria. The one or more bacterial powders and excipients are mixed as appropriate to form tablets. The tablets are taken daily for 3 months so that the total amount of bacteria taken is 1 ⁇ 10 6 to 1 ⁇ 10 12 cfu / kg body weight / day. By taking the tablet, a non-collagenous glycoprotein degradation effect can be expected.
  • bifidobacteria / oligosaccharide blended milk powder is prepared by adding 100 g of bacterial powder (1.8 ⁇ 10 11 cfu / g) obtained by concentrating and freeze-drying and then triturating with starch.
  • the prepared powdered milk was dissolved in water to prepare a formula having a total solid content of 14% (w / V) as a standard formula, the number of bifidobacteria in the formula was 2.7 ⁇ 10 9. cfu / 100 ml.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • Diabetes (AREA)
  • Hematology (AREA)
  • Wood Science & Technology (AREA)
  • Mycology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Ophthalmology & Optometry (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Polymers & Plastics (AREA)
  • Food Science & Technology (AREA)
  • Nutrition Science (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Pulmonology (AREA)
  • Epidemiology (AREA)
  • Emergency Medicine (AREA)
  • Endocrinology (AREA)
  • Obesity (AREA)
  • Toxicology (AREA)

Abstract

The present invention addresses the problem of providing an agent for degrading a non-collagenous glycoprotein, a composition for degrading a non-collagenous glycoprotein, a medicinal composition for degrading a non-collagenous glycoprotein and a food or beverage composition for degrading a non-collagenous glycoprotein, each being able to be easily produced and showing no side effects. To solve this problem, provided are an agent for degrading a non-collagenous glycoprotein, a composition for degrading a non-collagenous glycoprotein, a medicinal composition for degrading a non-collagenous glycoprotein and a food or beverage composition for degrading a non-collagenous glycoprotein, each comprising, as an active ingredient, a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium and/or treated cells of the bacterium.

Description

非コラーゲン性糖タンパク質分解用組成物Non-collagenous glycoprotein degradation composition
　本発明は、非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物、非コラーゲン性糖タンパク質分解用医薬組成物及び非コラーゲン性糖タンパク質分解用飲食品組成物に関する。 The present invention relates to a non-collagenous glycoprotein degrading agent or a non-collagenous glycoprotein degrading composition, a non-collagenous glycoprotein degrading pharmaceutical composition and a non-collagenous glycoprotein degrading food / beverage composition.
　非コラーゲン性糖タンパク質は、細胞や組織を囲む細胞外マトリックスの網目構造を形成する細胞接着分子の一種である。
　非コラーゲン性糖タンパク質のひとつであるフィブリノゲンは、血栓の形成に関与することが知られている。血管内皮が傷害、破綻すると、その部位に血小板の粘着凝集が生じ、血管収縮とともに血液凝固系が活性化されて、フィブリノゲンからフィブリンが生成し、フィブリン塊となって血栓を形成する。
　また、フィブロネクチンやラミニンは、眼の硝子体と網膜との界面の構成成分である非コラーゲン性糖タンパク質として知られている。眼の硝子体は、本来網膜と密着した状態にあるが、加齢に伴い硝子体がゲル状から液体状に変性し、網膜から剥離することが知られている（後部硝子体剥離）。後部硝子体剥離自体は病的な変化ではないが、硝子体と網膜が強固に結合していると、硝子体剥離時に網膜裂傷を引き起こし、重篤な視力障害を起こし得る。 Non-collagenous glycoprotein is a kind of cell adhesion molecule that forms a network structure of extracellular matrix surrounding cells and tissues.
Fibrinogen, one of non-collagenous glycoproteins, is known to be involved in thrombus formation. When the vascular endothelium is injured or ruptured, adhesion aggregation of platelets occurs at that site, the blood coagulation system is activated along with the vasoconstriction, and fibrin is generated from fibrinogen, forming a fibrin clot and forming a thrombus.
Fibronectin and laminin are also known as non-collagenous glycoproteins that are components of the interface between the vitreous body of the eye and the retina. The vitreous body of the eye is originally in close contact with the retina, but it is known that the vitreous body is denatured from a gel state to a liquid state with aging and peels from the retina (rear vitreous body peeling). The posterior vitreous detachment itself is not a pathological change, but if the vitreous and the retina are firmly bonded, a retinal tear may be caused at the time of vitreal detachment and serious visual impairment may be caused.
　一方で、プラスミンというセリンプロテアーゼが知られている。プラスミンは、エンドプロテアーゼに分類され、生体内において、通常は不活性型の前駆体であるプラスミノーゲンとして存在する。プラスミノーゲンアクチベーター（ＰＡ）によってプラスミノーゲンの特定のペプチド結合が分解され、活性化されて、プラスミンとして酵素活性を発揮する。プラスミノーゲンアクチベーターとして、血管内皮細胞から分泌される組織プラスミノーゲンアクチベーター（tissue plasminogen activator：ｔＰＡ）や、尿中に存在するウロキナーゼ型プラスミノーゲン活性化因子（ｕＰＡ）等が存在する。プラスミンは、フィブリノゲンやフィブロネクチン、ラミニンといった非コラーゲン性糖タンパク質を分解する活性を有することが知られている（非特許文献１）。 On the other hand, a serine protease called plasmin is known. Plasmin is classified as an endoprotease and exists in vivo as plasminogen, which is usually an inactive precursor. A specific peptide bond of plasminogen is decomposed and activated by plasminogen activator (PA) to exert enzyme activity as plasmin. Examples of plasminogen activators include tissue plasminogen activator (tPA) secreted from vascular endothelial cells and urokinase-type plasminogen activator (uPA) present in urine. Plasmin is known to have an activity of degrading non-collagenous glycoproteins such as fibrinogen, fibronectin and laminin (Non-patent Document 1).
　プラスミンが有するかかる非コラーゲン性糖タンパク質分解活性に基づく有用性から、プラスミンの種々の利用方法や製造方法が検討されている。
　例えば、プラスミンは、フィブリノゲン分解（fibrinogenolysis）を惹起してフィブリン塊を溶解する反応に関与する酵素であり、血液凝固系において形成された血栓を溶解する作用を有する。そのため、プラスミンは、フィブリン塊で形成される血栓によって引き起こされる虚血性障害の症状を予防又は軽減させる保存的治療法（線溶療法）に用いられる（非特許文献２）。
　また、プラスミンは、硝子体網膜界面の構成成分であるフィブロネクチン等に対しても分解活性を有することから、網膜から硝子体を切除する際の補助物質としても用いられている（特許文献１）。
　また、血漿または遺伝子組み換え体由来のプラスミノーゲンを精製した後、プラスミノーゲン活性化因子と接触させて当該プラスミノーゲンをプラスミンへ活性化した後、プラスミン含有画分をオメガ‐アミノ酸を含んでなるアフィニティークロマトグラフィーおよびゲルろ過クロマトグラフィーに付す、プラスミンの製造方法が報告されている（特許文献２）。
　また、ナットウキナーゼ等の細菌（納豆菌）由来のプラスミン活性を有する酵素が、血漿における線溶活性を増強することが報告されている（非特許文献３）。一方で、納豆中には、納豆菌が産生するビタミンＫが豊富に含まれており、納豆を摂取することによってビタミンＫが血中に移行して、一部の抗血栓薬の薬効を低下させることを示唆する報告がされている（非特許文献４及び非特許文献５）。
　また、これまでに、生体内に存在するビフィドバクテリウム属細菌の中には、細胞表層においてプラスミンノーゲンと結合し、宿主のプラスミノーゲンアクチベーターを利用して、細胞表層に結合したプラスミノーゲンを活性化してプラスミンに変換する活性を有するものが存在することが報告されている（非特許文献６）。 Due to the usefulness of plasmin based on such non-collagenous glycoproteolytic activity, various uses and production methods of plasmin have been studied.
For example, plasmin is an enzyme involved in a reaction that causes fibrinogenolysis and dissolves a fibrin clot, and has an action of dissolving a thrombus formed in the blood coagulation system. Therefore, plasmin is used in a conservative treatment method (fibrinolytic therapy) that prevents or reduces symptoms of ischemic injury caused by a thrombus formed by a fibrin clot (Non-patent Document 2).
Plasmin also has a degrading activity for fibronectin, which is a constituent component of the vitreous retinal interface, and is therefore used as an auxiliary substance when excising the vitreous from the retina (Patent Document 1).
In addition, after purifying plasma or a gene-derived plasminogen, contacting with a plasminogen activator to activate the plasminogen to plasmin, the plasmin-containing fraction contains omega-amino acids. There has been reported a method for producing plasmin that is subjected to affinity chromatography and gel filtration chromatography (Patent Document 2).
In addition, it has been reported that an enzyme having plasmin activity derived from bacteria (natto) such as nattokinase enhances fibrinolytic activity in plasma (Non-patent Document 3). On the other hand, natto contains abundant vitamin K produced by Bacillus natto, and by ingesting natto, vitamin K moves into the blood and reduces the efficacy of some antithrombotic drugs. There are reports that suggest this (Non-Patent Document 4 and Non-Patent Document 5).
In addition, to date, some Bifidobacterium bacteria that exist in the body bind to plasminogen on the cell surface and use the host plasminogen activator to bind to the cell surface. It has been reported that there is one having an activity of activating a gene and converting it into plasmin (Non-patent Document 6).
　しかしながら、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物が、プラスミノーゲンアクチベーター活性とプラスミン活性とを有し、それによって非コラーゲン性糖タンパク質分解効果を奏することは知られていない。 However, the Bifidobacterium genus, the culture of the bacterium, and / or the treated product of the bacterium have plasminogen activator activity and plasmin activity, thereby non-collagenous glycoprotein degradation effect It is not known to play.
特表２００６－５１８７０８号公報JP-T-2006-518708 特開２００７－６８４９７号公報JP 2007-68497 A
　前記プラスミンの製造方法は、プラスミン活性を損なわないように複雑な工程を経る必要がある。また、納豆由来のナットウキナーゼを利用する際には、納豆中のビタミンＫが血液の凝固を促進するという副作用が懸念される。
　本発明は、簡便に製造でき、副作用のない、非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物、非コラーゲン性糖タンパク質分解用医薬組成物及び非コラーゲン性糖タンパク質分解用飲食品組成物の提供を課題とする。 The method for producing plasmin needs to undergo complicated steps so as not to impair plasmin activity. Moreover, when using natto kinase derived from natto, there is a concern about the side effect that vitamin K in natto promotes blood coagulation.
The present invention is a non-collagenous glycoprotein degrading agent or a non-collagenous glycoprotein degrading composition, a non-collagenous glycoprotein degrading pharmaceutical composition, and a non-collagenous glycoprotein degrading food and beverage that can be produced simply and without side effects. An object is to provide a product composition.
　本発明者等が鋭意研究を進めた結果、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物が、プラスミノーゲンアクチベーター活性とプラスミン活性を有することに基づき、非コラーゲン性糖タンパク質分解効果を奏することを見出し、本発明を完成させた。 As a result of diligent research by the present inventors, the Bifidobacterium genus bacteria, the culture of the bacteria, and / or the treated product of the bacteria have plasminogen activator activity and plasmin activity. Based on this, it was found that a non-collagenous glycoprotein degradation effect was achieved, and the present invention was completed.
　すなわち、本発明は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とする非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物を提供する。 That is, the present invention relates to a non-collagenous glycoprotein degrading agent or a composition for degrading non-collagenous glycoprotein, comprising as an active ingredient a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium. Offer things.
　また、前記分解剤又は分解用組成物は、前記非コラーゲン性糖タンパク質が、フィブリン、フィブリノゲン、フィブロネクチン、ラミニン又はプラスミノーゲンであることを好ましい態様とする。 Further, the degradation agent or the composition for degradation is preferably a mode in which the non-collagenous glycoprotein is fibrin, fibrinogen, fibronectin, laminin or plasminogen.
　また、前記分解剤又は分解用組成物は、前記ビフィドバクテリウム属細菌が、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＡＴＣＣ１５７０７（Bifidobacterium longum subsp. longum ATCC15707）、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＡＴＣＣ　ＢＡＡ－９９９（Bifidobacterium longum subsp. longum ATCC BAA-999）（又はビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＢＢ５３６（ＮＩＴＥ　ＢＰ－０２６２１）（Bifidobacterium longum subsp. longum BB536（NITE BP-02621）））、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＡＴＣＣ１５６９７（Bifidobacterium longum subsp. infantis ATCC15697）、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＢＣＣＭ　ＬＭＧ２３７２８（Bifidobacterium longum subsp. infantis BCCM LMG23728）（又はビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＭ－６３（ＮＩＴＥ　ＢＰ－０２６２３）（Bifidobacterium longum subsp. infantis M-63（NITE BP-02623）））、ビフィドバクテリウム・ブレーベＡＴＣＣ１５７００（Bifidobacterium breve ATCC15700）、ビフィドバクテリウム・ブレーベＦＥＲＭ　ＢＰ－１１１７５（Bifidobacterium breve FERM BP-11175）、ビフィドバクテリウム・ブレーベＢＣＣＭ　ＬＭＧ２３７２９（Bifidobacterium breve BCCM LMG23729）（又はビフィドバクテリウム・ブレーベＭ－１６Ｖ（ＮＩＴＥ　ＢＰ－０２６２２）（Bifidobacterium breve M-16V（NITE BP-02622）））、ビフィドバクテリウム・ビフィダムＡＴＣＣ２９５２１（Bifidobacterium bifidum ATCC29521）、ビフィドバクテリウム・アドレセンティスＡＴＣＣ１５７０３（Bifidobacterium adolescentis ATCC15703）、ビフィドバクテリウム・アンギュラツムＡＴＣＣ２７５３５（Bifidobacterium angulatum ATCC27535）、ビフィドバクテリウム・デンティウムＤＳＭ２０４３６（Bifidobacterium dentium DSM20436）、ビフィドバクテリウム・シュードカテヌラータムＡＴＣＣ２７９１９（Bifidobacterium psudocatenulatum ATCC27919）、ビフィドバクテリウム・アニマリス・サブスピーシーズ・ラクティスＤＳＭ１０１４０（Bifidobacterium animalis subsp. lactis DSM10140）、ビフィドバクテリウム・アニマリス・サブスピーシーズ・アニマリスＡＴＣＣ２５５２７（Bifidobacterium animalis subsp. animalis ATCC25527）、ビフィドバクテリウム・シュードロンガム・サブスピーシーズ・グロボッサムＪＣＭ５８２０（Bifidobacterium pseudolongum subsp. globosum JCM5820）、ビフィドバクテリウム・シュードロンガム・サブスピーシーズ・シュードロンガムＡＴＣＣ２５５２６（Bifidobacterium pseudolongum subsp. pseudolongm ATCC25526）、及びビフィドバクテリウム・サーモフィルムＡＴＣＣ２５５２５（Bifidobacterium thermophilum ATCC25525）からなる群から選択される一又は複数であることを好ましい態様とする。 In addition, the decomposing agent or the decomposing composition is such that the Bifidobacterium genus bacterium is Bifidobacterium longum subspecies longum ATCC 15707 (Bifidobacterium longum subsp. Longum ATCC15707), Bifidobacterium longum subs. Spices Longum ATCC BAA-999 (Bifidobacterium longum subsp. Longum ATCC BAA-999) (or Bifidobacterium longum Subspecies longum BB536 (NITE BP-02621) (Bifidobacterium longum subsp. ))), Bifidobacterium longum subsp. Infatis ATCC 15697 (Bifidobacterium longum subsp. Infantis tATCC15697), Bifidobacterium longum subsp. Infantis BCCM MG23728 (Bifidobacterium longum subsp. Infantis BCCM LMG23728) (or Bifidobacterium longum subspecies Infantis M-63 (NITE BP-02623) (Bifidobacterium longum subsp. Infantis M-63 (NITE BP-02623)) ), Bifidobacterium breve ATCC15700 (Bifidobacterium breve ATCC15700), Bifidobacterium breve FERM BP-11175 (Bifidobacterium breve ＢFERM BP-11175), Bifidobacterium breve BCCM LMG23729 (Bifidobacterium breve GGCM Bifidobacterium breve M-16V (NITE BP-02622) (Bifidobacterium breve M-16V (NITE BP-02622))), Bifidobacterium bifidum ATCC29521 (Bifidobacterium bifidum ATCC29521), Bi Fidobacterium adrecentis ATCC15703 (Bifidobacterium adolescentis ATCC15703), Bifidobacterium angulatum ATCC 27535 (Bifidobacterium angulatum ATCC27535), Bifidobacterium dentium DSM20436 (Bifidobacterium dentium DSM20436), Bifidobacterium 19AT (Bifidobacterium psudocatenulatum ATCC27919), Bifidobacterium ド animalis subsp. Lactis DSM10140, Bifidobacterium ア ニ animalis subspicesanimalis ATCC25527 Bacteria Pseudolongum Subspecies Globosum JCM5 20 (Bifidobacterium pseudolongum subsp. Globosum JCM5820), Bifidobacterium pseudolongum subspices Pseudolongum ATCC 25526 (Bifidobacterium pseudolongum subsp. A preferred embodiment is one or more selected from the group consisting of:
　さらに、本発明は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とする非コラーゲン性糖タンパク質分解用医薬組成物を提供する。 Furthermore, the present invention provides a pharmaceutical composition for degrading non-collagenous glycoproteins comprising as an active ingredient a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium.
　前記医薬組成物は、脳梗塞、四肢動静脈血栓症、肺梗塞、脳静脈洞血栓、網膜剥離、網膜裂傷、硝子体出血、糖尿病性硝子体出血、増殖性糖尿、加齢黄斑変性、黄斑円孔、硝子体黄斑牽引、フィブリン沈着、網膜静脈閉塞症、網膜動脈閉塞症、緑内障又は網膜色素変性症の予防及び／又は治療に用いられることを好ましい態様とする。 The pharmaceutical composition is cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction, cerebral venous sinus thrombus, retinal detachment, retinal tear, vitreous hemorrhage, diabetic vitreous hemorrhage, proliferative diabetes, age-related macular degeneration, macular circle It is preferably used for prevention and / or treatment of pores, vitreous macular traction, fibrin deposition, retinal vein occlusion, retinal artery occlusion, glaucoma or retinitis pigmentosa.
　また、本発明は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とする非コラーゲン性糖タンパク質分解用飲食品組成物を提供する。 The present invention also provides a non-collagenized glycoprotein degrading food / beverage composition comprising as an active ingredient a Bifidobacterium genus bacterium, a culture of the bacterium, and / or a treated product of the bacterium.
　前記飲食品組成物は、プラスミノーゲンを含むことを好ましい態様とする。 The food / beverage product composition preferably contains plasminogen.
　さらに、本発明は、ビフィドバクテリウム属細菌を培養する工程、及び前記培養後の培養物中に産生されたプラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質を回収する工程を含む、プラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質の製造方法を提供する。 Furthermore, the present invention includes a step of culturing Bifidobacterium, and a step of recovering a protein having plasminogen activator activity and / or plasmin activity produced in the culture after the culture. Provided is a method for producing a protein having plasminogen activator activity and / or plasmin activity.
　さらに、本発明は、プラスミノーゲンを含む培地中でビフィドバクテリウム属細菌を培養する工程、及び前記培養後の培養物中に産生されたプラスミンを回収する工程を含む、プラスミンの製造方法を提供する。 Furthermore, the present invention provides a method for producing plasmin, comprising a step of culturing Bifidobacterium in a medium containing plasminogen, and a step of recovering plasmin produced in the culture after the culture. provide.
　さらに、本発明は、非コラーゲン性糖タンパク質分解用組成物の製造における、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物の使用を提供する。
　また、本発明は、非コラーゲン性糖タンパク質分解に用いられるビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を提供する。
　また、本発明は、非コラーゲン性糖タンパク質分解のためのビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物の使用を提供する。
　また、本発明は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を哺乳動物に投与する段階を含む、非コラーゲン性糖タンパク質を分解する方法を提供する。 Furthermore, the present invention provides use of a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium in the manufacture of a composition for degrading non-collagenous glycoprotein.
The present invention also provides a Bifidobacterium genus used for non-collagenous glycoprotein degradation, a culture of the bacterium, and / or a treated product of the bacterium.
The present invention also provides use of a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium for non-collagenous glycoprotein degradation.
The present invention also provides a method for degrading non-collagenous glycoprotein, comprising the step of administering a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium to a mammal. To do.
　本発明によれば、簡便に製造でき、副作用のない、非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物、非コラーゲン性糖タンパク質分解用医薬組成物及び非コラーゲン性糖タンパク質分解用飲食品組成物を提供できる。 According to the present invention, a non-collagenous glycoprotein degrading agent or a non-collagenous glycoprotein degrading composition, a non-collagenous glycoprotein degrading pharmaceutical composition, and a non-collagenous glycoprotein degrading agent that can be easily produced and have no side effects. The food-drinks composition can be provided.
　次に、本発明の好ましい実施形態について説明する。ただし、本発明は以下の好ましい実施形態に限定されず、本発明の範囲内で自由に変更することができる。尚、本明細書において百分率は特に断りのない限り質量による表示である。 Next, a preferred embodiment of the present invention will be described. However, the present invention is not limited to the following preferred embodiments, and can be freely changed within the scope of the present invention. In the present specification, percentages are expressed by mass unless otherwise specified.
＜非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物＞
　本発明に係る非コラーゲン性糖タンパク質分解剤は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分として含む。尚、本発明に係る非コラーゲン性糖タンパク質分解剤は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分として含むものであって、他の成分を含むことを妨げるものではない。すなわち、本発明に係る非コラーゲン性糖タンパク質分解剤は、非コラーゲン性糖タンパク質分解用組成物と同等である。よって、本発明の他の態様は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とする非コラーゲン性糖タンパク質分解用組成物である。 <Non-collagenous glycoprotein degrading agent or non-collagenous glycoprotein degrading composition>
The non-collagenous glycoprotein degrading agent according to the present invention contains a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient. The non-collagenous glycoprotein degrading agent according to the present invention contains a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient. It does not prevent the inclusion of ingredients. That is, the non-collagenous glycoprotein degrading agent according to the present invention is equivalent to the non-collagenous glycoprotein degrading composition. Therefore, another aspect of the present invention is a composition for degrading non-collagenous glycoproteins comprising a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient.
　また、該組成物中のビフィドバクテリウム属細菌の含有量は、好ましくは１×１０６～１×１０１２ＣＦＵ／ｇ又は１×１０６～１×１０１２ＣＦＵ／ｍＬであり、より好ましくは１×１０７～１×１０１１ＣＦＵ／ｇ又は１×１０７～１×１０１１ＣＦＵ／ｍＬであり、さらに好ましくは１×１０８～１×１０１０ＣＦＵ／ｇ又は１×１０８～１×１０１０ＣＦＵ／ｍＬである。該細菌が死菌の場合、ＣＦＵは個細胞（cells）と置き換えることができる。 The content of Bifidobacterium in the composition is preferably 1 × 10 6 to 1 × 10 12 CFU / g or 1 × 10 6 to 1 × 10 12 CFU / mL, more preferably Is 1 × 10 7 to 1 × 10 11 CFU / g or 1 × 10 7 to 1 × 10 11 CFU / mL, more preferably 1 × 10 8 to 1 × 10 10 CFU / g or 1 × 10 8 1 × 10 10 CFU / mL. If the bacterium is dead, the CFU can be replaced with cells.
　当該ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物は、プラスミノーゲンアクチベーター活性とプラスミン活性とを有し、これらに基づいて、非コラーゲン性糖タンパク質の分解効果を示すものである。
　したがって、本発明においては、非特許文献６のように、細胞表層にプラスミンノーゲンが結合していなくてもプラスミノーゲンをプラスミンに変換することができる。すなわち、本発明では、ビフィドバクテリウム属細菌の細胞表層にプラスミンノーゲンを結合させる工程を必要としない。 The Bifidobacterium bacterium, the culture of the bacterium, and / or the treated product of the bacterium have plasminogen activator activity and plasmin activity, and based on these, non-collagenous glycoprotein This shows the decomposition effect.
Therefore, in the present invention, plasminogen can be converted to plasmin even if plasminogen is not bound to the cell surface as in Non-Patent Document 6. That is, the present invention does not require a step of binding plasminogen to the cell surface of Bifidobacterium.
　また、本発明の別の側面は、本発明のビフィドバクテリウム属細菌を組成物原料に添加する工程を含む、非コラーゲン性糖タンパク質分解用組成物の製造方法である。当該製造方法は、本発明のビフィドバクテリウム属細菌を飲食品組成物原料に添加する工程を含む、飲食品組成物の製造方法を含む。また、当該製造方法は、ビフィドバクテリウム属細菌を医薬品組成物原料（例えば、基剤等）に添加する工程を含む、医薬組成物の製造方法を含む。
　当該非コラーゲン性糖タンパク質分解用組成物の製造方法に使用されるビフィドバクテリウム属細菌は、生菌であっても死菌であってもよく、生菌と死菌との両方を含むものでもよい。さらに、ビフィドバクテリウム属細菌の培養物であってもよく、菌体処理物であってもよい。また、当該非コラーゲン性糖タンパク質分解用組成物の製造方法においては、ビフィドバクテリウム属細菌は、培養し濃縮又は凍結乾燥した後に組成物原料に添加されてもよく、ビフィドバクテリウム属細菌を組成物原料に添加した後に該ビフィドバクテリウム属細菌を培養してもよい。 Another aspect of the present invention is a method for producing a composition for degrading non-collagenous glycoprotein, comprising a step of adding the Bifidobacterium of the present invention to a composition raw material. The said manufacturing method contains the manufacturing method of the food-drinks composition including the process of adding the Bifidobacterium genus bacteria of this invention to the food-drinks composition raw material. Moreover, the said manufacturing method contains the manufacturing method of pharmaceutical composition including the process of adding Bifidobacterium genus bacteria to pharmaceutical composition raw materials (for example, base etc.).
The Bifidobacterium genus bacterium used in the method for producing the composition for degrading non-collagenous glycoprotein may be live or dead, and includes both live and dead But you can. Furthermore, it may be a culture of a Bifidobacterium bacterium or a cell-treated product. Further, in the method for producing a composition for degrading non-collagenous glycoprotein, the Bifidobacterium bacterium may be added to the composition raw material after culturing, concentrating or freeze-drying, and the Bifidobacterium bacterium The Bifidobacterium may be cultured after adding to the composition raw material.
（１）ビフィドバクテリウム属細菌
　本発明に用いることができるビフィドバクテリウム属細菌には、本発明の効果を損なわない限り、公知のビフィドバクテリウム属細菌を用いることができる。例えば、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガム（Bifidobacterium longum subsp. longum）、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティス（Bifidobacterium longum subsp. infantis）、ビフィドバクテリウム・ブレーベ（Bifidobacterium breve）、ビフィドバクテリウム・ビフィダム（Bifidobacterium bifidum）、ビフィドバクテリウム・アドレセンティス（Bifidobacterium adolescentis）、ビフィドバクテリウム・アンギュラツム（Bifidobacterium angulatum）、ビフィドバクテリウム・デンティウム（Bifidobacterium dentium）、ビフィドバクテリウム・シュードカテヌラータム（Bifidobacterium psudocatenulatum）、ビフィドバクテリウム・アニマリス・サブスピーシーズ・ラクティス（Bifidobacterium animalis subsp. lactis）、ビフィドバクテリウム・アニマリス・サブスピーシーズ・アニマリス（Bifidobacterium animalis subsp. animalis）、ビフィドバクテリウム・シュードロンガム・サブスピーシーズ・グロボッサム（Bifidobacterium pseudolongum subsp. globosum）、ビフィドバクテリウム・シュードロンガム・サブスピーシーズ・シュードロンガム（Bifidobacterium pseudolongum subsp. pseudolongm）、ビフィドバクテリウム・サーモフィルム（Bifidobacterium thermophilum）等が例示できる。なお、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムは、単にビフィドバクテリウム・ロンガムと表記される場合もある。また、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスは、単にビフィドバクテリウム・インファンティスと表記される場合もある。 (1) Bifidobacterium genus Bacteria belonging to the genus Bifidobacterium can be used in the genus Bifidobacterium, as long as the effects of the present invention are not impaired. For example, Bifidobacterium longum subsp. Longum, Bifidobacterium longum subsp. Infantis, Bifidobacterium breve (Bifidobacterium breve), Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium angulatum, Bifidobacterium dentium, Bifidobacterium adolescentis, Bifidobacterium angulatum Bifidobacterium psudocatenulatum, Bifidobacterium animalis subsp. Lactis, Bifidobacterium anima Bifidobacterium animalis subsp. Animalis, Bifidobacterium pseudolongum subsp. Globosum, Bifidobacterium pseudolongum subsp. Globosum (Bifidobacterium pseudolongum subsp. Pseudolongm), Bifidobacterium thermophilum, etc. can be illustrated. The Bifidobacterium longum subspecies longum may be simply referred to as Bifidobacterium longum. In addition, Bifidobacterium longum sub-species Infantis is sometimes simply referred to as Bifidobacterium Infantis.
　具体的には、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＡＴＣＣ１５７０７（Bifidobacterium longum subsp. longum ATCC15707）、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＡＴＣＣ　ＢＡＡ－９９９（Bifidobacterium longum subsp. longum ATCC BAA-999）又はビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＢＢ５３６（ＮＩＴＥ　ＢＰ－０２６２１）（Bifidobacterium longum subsp. longum BB536（NITE BP-02621））、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＡＴＣＣ１５６９７（Bifidobacterium longum subsp. infantis ATCC15697）、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＢＣＣＭ　ＬＭＧ２３７２８（Bifidobacterium longum subsp. infantis BCCM LMG23728）又はビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＭ－６３（ＮＩＴＥ　ＢＰ－０２６２３）（Bifidobacterium longum subsp. infantis M-63（NITE BP-02623））、ビフィドバクテリウム・ブレーベＡＴＣＣ１５７００（Bifidobacterium breve ATCC15700）、ビフィドバクテリウム・ブレーベＦＥＲＭ　ＢＰ－１１１７５（Bifidobacterium breve FERM BP-11175）、ビフィドバクテリウム・ブレーベＢＣＣＭ　ＬＭＧ２３７２９（Bifidobacterium breve BCCM LMG23729）又はビフィドバクテリウム・ブレーベＭ－１６Ｖ（ＮＩＴＥ　ＢＰ－０２６２２）（Bifidobacterium breve M-16V（NITE BP-02622））、ビフィドバクテリウム・ビフィダムＡＴＣＣ２９５２１（Bifidobacterium bifidum ATCC29521）、ビフィドバクテリウム・アドレセンティスＡＴＣＣ１５７０３（Bifidobacterium adolescentis ATCC15703）、ビフィドバクテリウム・アンギュラツムＡＴＣＣ２７５３５（Bifidobacterium angulatum ATCC27535）、ビフィドバクテリウム・デンティウムＤＳＭ２０４３６（Bifidobacterium dentium DSM20436）、ビフィドバクテリウム・シュードカテヌラータムＡＴＣＣ２７９１９（Bifidobacterium psudocatenulatum ATCC27919）、ビフィドバクテリウム・アニマリス・サブスピーシーズ・ラクティスＤＳＭ１０１４０（Bifidobacterium animalis subsp. lactis DSM10140）、ビフィドバクテリウム・アニマリス・サブスピーシーズ・アニマリスＡＴＣＣ２５５２７（Bifidobacterium animalis subsp. animalis ATCC25527）、ビフィドバクテリウム・シュードロンガム・サブスピーシーズ・グロボッサムＪＣＭ５８２０（Bifidobacterium pseudolongum subsp. globosum JCM5820）、ビフィドバクテリウム・シュードロンガム・サブスピーシーズ・シュードロンガムＡＴＣＣ２５５２６（Bifidobacterium pseudolongum subsp. pseudolongm ATCC25526）、ビフィドバクテリウム・サーモフィルムＡＴＣＣ２５５２５（Bifidobacterium thermophilum ATCC25525）等が例示できる。ビフィドバクテリウム属細菌は、一種のみを用いてもよいし、任意の二種以上を用いてもよい。 Specifically, Bifidobacterium longum subsp. Longum ATCC 15707 (Bifidobacterium longum subsp. Longum ATCC15707), Bifidobacterium longum subspice longum ATCC BAA-999 (Bifidobacterium longum subsp. Longlong ATCC BABA- 999) or Bifidobacterium longum subspecies longis BB536 (NITE BP-02621) (Bifidobacterium longum subsp. Longum BB536 (NITE BP-02621)), Bifidobacterium longum subspecies Infatis ATCC 15697 (Bifidobacterium longum subsp. Infantis ATCC15697), Bifidobacterium longum subspices Infantis BCCM LMG23728 (Bifidobacterium longum subsp. Infantis BCCM LMG23728) or Fidobacterium longum subspecies Infantis M-63 (NITE BP-02623) (Bifidobacterium longum subsp. Infantis M-63 (NITE BP-02623)), Bifidobacterium breve ATCC15700 (Bifidobacterium breve ATCC15700) Bifidobacterium breve FERM BP-11175 (Bifidobacterium breve FERM BP-11175), Bifidobacterium breve BCCM LMG23729 (Bifidobacterium breve bc BCCM LMG23729) or Bifidobacterium breve M-16V (NITE BP-0222) (Bifidobacterium breve M-16V (NITE BP-02622)), Bifidobacterium bifidum ATCC29521 (Bifidobacteriumifbifidum ATCC29521), Bifidobacterium adrecentis ATCC15703 (Bifidobacterium adol escentis ATCC15703), Bifidobacterium angulatum ATCC27535, Bifidobacterium dentium DSM20436 (Bifidobacterium dentium DSM20436), Bifidobacterium カ テ dentium ＡＴATCC27919 Um animalis subspecies lactis DSM10140 (Bifidobacterium animalis subsp.・ Grobosum JCM5820 (Bifidobacterium pseudolongum subsp. Globosum JCM5820), Bifidobacterium Pseudolongum subspecies pseudolongum ATCC25526 (Bifidobacterium pseudolongum subsp. Pseudolongm ATCC25526), Bifidobacterium thermofilm ATCC25525 (Bifidobacterium thermophilum ATCC25525) and the like can be exemplified. As the Bifidobacterium, only one kind may be used, or any two or more kinds may be used.
　ＡＴＣＣ番号が付与された細菌は、アメリカン・タイプ・カルチャー・コレクション（住所：12301  Parklawn Drive, Rockville, Maryland 20852, United States of America）から入手することができる。
　ＢＣＣＭ番号が付与された細菌は、ベルギーの保存機関であるBelgian  Coordinated Collections of Microorganisms（住所：ベルギー、B-1000 ブリュッセル シアンス通り（ウェーテンスカップ通り）８）から入手することができる。
　ＦＥＲＭ　ＢＰ－１１１７５の受託番号が付与された細菌は、２００９年８月２５日付で、独立行政法人産業技術総合研究所特許生物寄託センター（現　独立行政法人製品評価技術基盤機構特許生物寄託センター、〒292-0818　千葉県木更津市かずさ鎌足2-5-8 120号室）にブダペスト条約に基づく国際寄託がなされている。
　ＤＳＭ番号が付与された細菌は、Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH（住所：Inhoffenstraβe 7B, 38124 Braunschweig, Germany）から入手することができる。
　ＪＣＭ番号が付与された細菌は、Japan Collection of Microorganisms（国立研究開発法人理化学研究所バイオリソースセンター微生物材料開発室、郵便番号：305-0074、住所：茨城県つくば市高野台3-1-1）から入手することができる。 Bacteria with ATCC numbers can be obtained from the American Type Culture Collection (address: 12301 Parklawn Drive, Rockville, Maryland 20852, United States of America).
Bacteria to which a BCCM number is assigned can be obtained from Belgian Coordinated Collections of Microorganisms (Address: B-1000 Brussels, Ciens Street (Wetens Cup Street) 8), Belgium.
Bacteria to which the FERM BP-11175 accession number has been assigned were issued on August 25, 2009 at the National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center (currently the National Institute for Product Evaluation Technology Patent Biological Deposit Center, 292-0818 2-5-8 Kazusa-Kamashita, Kisarazu City, Chiba Prefecture Room 120) has been deposited internationally based on the Budapest Treaty.
Bacteria with DSM numbers can be obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (address: Inhoffenstraβe 7B, 38124 Braunschweig, Germany).
Bacteria with JCM numbers are from the Japan Collection of Microorganisms (National Institute for Physical and Chemical Research, Bioresource Center, Microbial Materials Development Office, Postal Code: 305-0074, Address: 3-1-1 Takanodai, Tsukuba City, Ibaraki Prefecture) It can be obtained.
　尚、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＡＴＣＣ　ＢＡＡ－９９９は、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＢＢ５３６（ＮＩＴＥ　ＢＰ－０２６２１）と同一の細菌であり、好ましい態様においては、いずれの細菌を用いてもよい。ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＢＢ５３６（ＮＩＴＥ　ＢＰ－０２６２１）は、２０１８年１月２６日に、独立行政法人製品評価技術基盤機構特許微生物寄託センター（〒292-0818　千葉県木更津市かずさ鎌足2-5-8 122号室）に、ＮＩＴＥ　ＢＰ－０２６２１の受託番号で、ブダペスト条約に基づく国際寄託がなされたものである。 Bifidobacterium longum sub-species longum ATCC BAA-999 is the same bacterium as Bifidobacterium longum sub-species longum BB536 (NITE BP-02621). Bacteria may be used. Bifidobacterium longum subspecies longum BB536 (NITE BP-02621) was established on January 26, 2018, by the National Institute for Product Evaluation Technology Patent Microorganism Depositary Center (Kazusa, Kisarazu City, Chiba Prefecture 292-0818). International deposit based on the Budapest Treaty was made in Nama BP-02621 under No. 122 (Kamaashi 2-5-8 122).
　また、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＢＣＣＭ　ＬＭＧ２３７２８は、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＭ－６３（ＮＩＴＥ　ＢＰ－０２６２３）と同一の細菌であり、好ましい態様においては、いずれの細菌を用いてもよい。ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＭ－６３（ＮＩＴＥ　ＢＰ－０２６２３）は、２０１８年１月２６日に、独立行政法人製品評価技術基盤機構特許微生物寄託センター（〒292-0818　千葉県木更津市かずさ鎌足2-5-8 122号室）に、ＮＩＴＥ　ＢＰ－０２６２３の受託番号で、ブダペスト条約に基づく国際寄託がなされたものである。 In addition, Bifidobacterium longum subspecies Infatis BCCM LMG23728 is the same bacterium as Bifidobacterium longum subspecies Infantis M-63 (NITE BP-02623), which is a preferred embodiment. In, any bacteria may be used. Bifidobacterium longum subspecies Infantis M-63 (NITE BP-02623) was established on January 26, 2018, by the National Institute of Technology and Evaluation of the National Institute of Technology and Evaluation (JAPAN) In the prefecture, Kisarazu City, Kazusa Kamashi, 2-5-8 122), the deposit number of NITE BP-02623 was made based on the Budapest Treaty.
　また、ビフィドバクテリウム・ブレーベＢＣＣＭ　ＬＭＧ２３７２９は、ビフィドバクテリウム・ブレーベＭ－１６Ｖ（ＮＩＴＥ　ＢＰ－０２６２２）と同一の細菌であり、好ましい態様においては、いずれの細菌を用いてもよい。ビフィドバクテリウム・ブレーベＭ－１６Ｖ（ＮＩＴＥ　ＢＰ－０２６２２）は、２０１８年１月２６日に、独立行政法人製品評価技術基盤機構特許微生物寄託センター（〒292-0818　千葉県木更津市かずさ鎌足2-5-8 122号室）に、ＮＩＴＥ　ＢＰ－０２６２２の受託番号で、ブダペスト条約に基づく国際寄託がなされたものである。 Also, Bifidobacterium breve BCCM LMG23729 is the same bacterium as Bifidobacterium breve M-16V (NITE BP-02622), and any bacterium may be used in a preferred embodiment. Bifidobacterium breve M-16V (NITE BP-02622) was issued on January 26, 2018, by the National Institute for Product Evaluation Technology Patent Microorganisms Depositary Center (Kazusa Kamashi 2, Kisarazu City, Chiba Prefecture 292-0818) -5-8 Room 122), the deposit number of NITE BP-02622, which was internationally deposited under the Budapest Treaty.
　本発明のビフィドバクテリウム属細菌は、前記寄託菌に制限されず、前記寄託菌と実質的に同等の細菌であってもよい。前記寄託菌と実質的に同等の細菌とは、前記寄託菌と同属又は同種の細菌であって、前記細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を対象に摂取させたとき又は投与したときに、該対象において非コラーゲン性糖タンパク質を分解することができ、その１６ＳｒＲＮＡ遺伝子の塩基配列が、前記寄託菌の１６ＳｒＲＮＡ遺伝子の塩基配列と９８％以上、好ましくは９９％以上、より好ましくは１００％の相同性を有し、かつ、好ましくは、前記相同性に加えて、前記寄託菌と同一の菌学的性質を有する細菌である。また、本発明のビフィドバクテリウム属細菌は、本発明の効果が損なわれない限り、前記寄託菌、又はそれと実質的に同等の細菌から、変異処理、遺伝子組換え、自然変異株の選択等によって育種された細菌であってもよい。 The Bifidobacterium genus bacterium of the present invention is not limited to the deposited bacterium, and may be a bacterium substantially equivalent to the deposited bacterium. The bacterium substantially equivalent to the deposited bacterium is a bacterium belonging to the same genus or the same species as the deposited bacterium, and allows the subject to ingest the bacterium, a culture of the bacterium, and / or a treated product of the bacterium. Or when administered, non-collagenous glycoprotein can be degraded in the subject, and the base sequence of the 16S rRNA gene is 98% or more, preferably 99% or more of the base sequence of the 16S rRNA gene of the deposited bacterium. More preferably, the bacterium has a homology of 100%, and preferably a bacterium having the same mycological properties as the deposited bacterium in addition to the homology. In addition, as long as the effects of the present invention are not impaired, the Bifidobacterium genus bacterium of the present invention is selected from the deposited bacterium or a bacterium substantially equivalent thereto, mutation treatment, genetic recombination, selection of a natural mutant, etc. Bacteria bred by may be used.
　これらの中でも、本発明においては、ヒト常在性のビフィドバクテリウム属細菌を用いることが好ましい。ビフィドバクテリウム属細菌は、ヒト以外にも昆虫や小動物にも生息しているが、菌種によって生息域（宿主）が限定されており、主にヒトに生息しているビフィドバクテリウム属細菌はヒト常在性ビフィドバクテリウム属細菌（ＨＲＢ：Human-residential bifidobacteria）と呼ばれ、それ以外はｎｏｎ‐ＨＲＢと呼ばれる。ＨＲＢとしては、ビフィドバクテリウム・ロンガム（Bifidobacterium longum）、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティス（Bifidobacterium longum subsp. infantis）、ビフィドバクテリウム・ブレーベ（Bifidobacterium breve）、ビフィドバクテリウム・ビフィダム（Bifidobacterium bifidum）、ビフィドバクテリウム・アドレセンティス（Bifidobacterium adolescentis）、ビフィドバクテリウム・アンギュラツム（Bifidobacterium angulatum）、ビフィドバクテリウム・デンティウム（Bifidobacterium dentium）、ビフィドバクテリウム・シュードカテヌラータム（Bifidobacterium psudocatenulatum）等が例示できる。
　さらに、本発明においては、ＨＲＢの中でも乳幼児由来のビフィドバクテリウム属細菌がより好ましい。乳幼児由来のＨＲＢとしては、ビフィドバクテリウム・ロンガム（Bifidobacterium longum）、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティス（Bifidobacterium longum subsp. infantis）、ビフィドバクテリウム・ブレーベ（Bifidobacterium breve）、ビフィドバクテリウム・ビフィダム（Bifidobacterium bifidum）等が例示できる。 Among these, in the present invention, it is preferable to use human resident Bifidobacterium. Bifidobacterium spp. Live in insects and small animals in addition to humans, but the habitat (host) is limited by the species, and Bifidobacterium spp. The bacterium is called a human resident Bifidobacterium (HRB), otherwise it is called non-HRB. HRB includes Bifidobacterium longum, Bifidobacterium longum subsp. Infantis, Bifidobacterium breve, Bifidobacterium Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium angulatum, Bifidobacterium dentium, Bifidobacterium pseudocatenu Examples include ratum (Bifidobacterium psudocatenulatum) and the like.
Furthermore, in the present invention, Bifidobacterium bacteria derived from infants are more preferable among HRBs. Examples of HRB derived from infants include Bifidobacterium longum, Bifidobacterium longum subsp. Infantis, Bifidobacterium breve, Examples thereof include Bifidobacterium bifidum.
　本発明に用いられるビフィドバクテリウム属細菌は、上記ビフィドバクテリウム属細菌と同等以上の非コラーゲン性糖タンパク質分解効果を有する限り、上記ビフィドバクテリウム属細菌の変異株であってもよい。ある変異株が上記ビフィドバクテリウム属細菌と「同等以上の非コラーゲン性糖タンパク質分解効果」を有するか否かは、例えば、後述するビフィドバクテリウム属細菌のプラスミン活性の測定や、プラスミノーゲンアクチベーター活性の測定により確認できる。
　このような変異株は、上記ビフィドバクテリウム属細菌に非人為的に変異が導入されることで構築されてもよい。また、ＵＶ等の変異原を用いた処理により上記細菌に変異を導入して構築してもよく、種々の遺伝子操作法により上記細菌に変異を導入して構築してもよい。 The Bifidobacterium bacterium used in the present invention may be a mutant strain of the Bifidobacterium bacterium as long as it has a non-collagenous glycoprotein degrading effect equivalent to or better than the Bifidobacterium bacterium. . Whether or not a mutant strain has a “non-collagenous glycoprotein degradation effect equal to or higher than that of the above-mentioned Bifidobacterium genus” can be determined, for example, by measuring the plasmin activity of Bifidobacterium genus described later, It can be confirmed by measuring gen activator activity.
Such a mutant strain may be constructed by introducing a mutation artificially into the Bifidobacterium. In addition, it may be constructed by introducing mutation into the bacterium by treatment with a mutagen such as UV, or may be constructed by introducing mutation into the bacterium by various gene manipulation methods.
　本発明において、プラスミン活性は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物とプラスミンに特異的な蛍光基質とを混合し、培養した培養物の蛍光強度を測定することにより確認することができる。すなわち、前記培養物の蛍光強度から、前記蛍光基質由来の蛍光物質の濃度と蛍光強度との関係を示す検量線を用いて、前記培養物中の蛍光物質濃度を算出し、さらに、１分間に産生される蛍光物質１ｎｍｏｌを酵素１単位（Ｕ）として、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物の酵素活性（プラスミン活性）を決定することができる。
　例えば、ビフィドバクテリウム属細菌の培養物から菌体を分離してＰＢＳ溶液に懸濁して懸濁液を調製し、当該懸濁液を希釈して濁度（ＯＤ６００）を０．１になるように調整した後、プラスミンに特異的な蛍光基質であるＢｏｃ‐Ｖａｌ‐Ｌｅｕ‐Ｌｙｓ‐ＡＭＣ（ＢＡＣＨＥＭ社製）を添加して６０分間、３７℃で嫌気的に培養し、培養終了後、ｍｉｃｒｏｐｌａｔｅ　ｒｅａｄｅｒ　ＳＨ‐９０００（Corona Electric社製）等の蛍光測定装置により、励起波長３８０ｎｍ、測定波長４６０ｎｍで測定した培養物の蛍光強度から、ビフィドバクテリウム属細菌の酵素活性（プラスミン活性）を決定することができる。
　本発明において、当該プラスミン活性は、好ましくは４０μＵ以上、より好ましくは８０μＵ以上、さらに好ましくは１００μＵ以上であれば、本発明の非コラーゲン性糖タンパク質分解効果を好適に発揮することができる。 In the present invention, the plasmin activity is the fluorescence of a culture obtained by mixing a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium with a fluorescent substrate specific to plasmin, and culturing the mixture. This can be confirmed by measuring the strength. That is, the fluorescent substance concentration in the culture is calculated from the fluorescent intensity of the culture using a calibration curve indicating the relationship between the fluorescent substance concentration and the fluorescent intensity derived from the fluorescent substrate, and further, in one minute. Determination of enzyme activity (plasmin activity) of a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium, with 1 nmol of the fluorescent substance produced as 1 unit of enzyme (U) it can.
For example, cells are separated from a culture of Bifidobacterium and suspended in a PBS solution to prepare a suspension, and the suspension is diluted to a turbidity (OD600) of 0.1. After adjusting as described above, Boc-Val-Leu-Lys-AMC (manufactured by BACHEM), which is a fluorescent substrate specific for plasmin, was added and cultured anaerobically at 37 ° C. for 60 minutes. The enzyme activity (plasmin activity) of Bifidobacterium is determined from the fluorescence intensity of the culture measured at an excitation wavelength of 380 nm and a measurement wavelength of 460 nm with a fluorescence measuring device such as reader SH-9000 (Corona Electric). be able to.
In the present invention, when the plasmin activity is preferably 40 μU or more, more preferably 80 μU or more, and even more preferably 100 μU or more, the non-collagenous glycoprotein degradation effect of the present invention can be suitably exhibited.
　また、本発明において、プラスミノーゲンアクチベーター活性は、プラスミンに特異的な蛍光基質ではなく、プラスミノーゲンアクチベーターに特異的な蛍光基質としてＺ‐Ｇｌｙ‐Ｇｌｙ‐Ａｒｇ‐ＡＭＣ・ＨＣｌ（ＢＡＣＨＥＭ社製）を用いること以外は上記プラスミン活性の場合と同様にして決定することができる。
　本発明において、当該プラスミノーゲンアクチベーター活性は、好ましくは１０μＵ以上、より好ましくは２０μＵ以上、さらに好ましくは５０μＵ以上であれば、本発明の非コラーゲン性糖タンパク質分解効果を好適に発揮することができる。 In the present invention, plasminogen activator activity is not a plasmin-specific fluorescent substrate, but a plasminogen activator-specific fluorescent substrate as Z-Gly-Gly-Arg-AMC.HCl (BACHEM). It can be determined in the same manner as in the case of the plasmin activity except that the product is used.
In the present invention, if the plasminogen activator activity is preferably 10 μU or more, more preferably 20 μU or more, and even more preferably 50 μU or more, the non-collagenous glycoprotein degradation effect of the present invention can be suitably exhibited. it can.
　本発明に用いられるビフィドバクテリウム属細菌、及び前記細菌の培養物は、常法によりビフィドバクテリウム属細菌を培養することにより容易に取得できる。培養方法は、ビフィドバクテリウム属細菌が増殖できる限り特に限定されず、細菌の性質に応じた適当な条件下で培養を行うことができる。例えば、培養温度は２５～５０℃でよく、３５～４２℃であることが好ましい。また培養は嫌気条件下で行うことが好ましく、例えば、炭酸ガス等の嫌気ガスを通気しながら培養することができる。また、液体静置培養等の微好気条件下で培養してもよい。 The Bifidobacterium bacterium used in the present invention and the culture of the bacterium can be easily obtained by culturing the Bifidobacterium bacterium by a conventional method. The culture method is not particularly limited as long as Bifidobacterium can grow, and culture can be performed under appropriate conditions according to the nature of the bacteria. For example, the culture temperature may be 25 to 50 ° C., preferably 35 to 42 ° C. Moreover, it is preferable to perform culture | cultivation on anaerobic conditions, for example, it can culture | cultivate, ventilating anaerobic gas, such as a carbon dioxide gas. Moreover, you may culture | cultivate on microaerobic conditions, such as liquid stationary culture.
　本発明に用いられるビフィドバクテリウム属細菌を培養する培地としては、特に限定されず、ビフィドバクテリウム属細菌の培養に通常用いられる培地を用いることができる。すなわち、炭素源としては、例えば、グルコース、ガラクトース、ラクトース、アラビノース、マンノース、スクロース、デンプン、デンプン加水分解物、廃糖蜜等の糖類を資化性に応じて使用できる。窒素源としては、例えば、アンモニア、硫酸アンモニウム、塩化アンモニウム、硝酸アンモニウム等のアンモニウム塩類や硝酸塩類を使用できる。また、無機塩類としては、例えば、塩化ナトリウム、塩化カリウム、リン酸カリウム、硫酸マグネシウム、塩化カルシウム、硝酸カルシウム、塩化マンガン、硫酸第一鉄等を用いることができる。また、ペプトン、大豆粉、脱脂大豆粕、肉エキス、酵母エキス等の有機成分を用いてもよい。 The medium for culturing Bifidobacterium used in the present invention is not particularly limited, and a medium usually used for culturing Bifidobacterium can be used. That is, as the carbon source, for example, saccharides such as glucose, galactose, lactose, arabinose, mannose, sucrose, starch, starch hydrolyzate, and molasses can be used depending on utilization. As the nitrogen source, for example, ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, and ammonium nitrate, and nitrates can be used. Examples of inorganic salts that can be used include sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, and ferrous sulfate. Organic components such as peptone, soybean powder, defatted soybean meal, meat extract, yeast extract and the like may also be used.
　本発明に用いられるビフィドバクテリウム属細菌は、これまでの説明のとおり、細菌そのもの、前記細菌の培養物、又は前記細菌の菌体処理物の形態で用いることができる。すなわち、ビフィドバクテリウム属細菌そのものでもよく、ビフィドバクテリウム属細菌を培養した後、得られた培養物をそのまま用いてもよく、希釈又は濃縮して用いてもよい。また、本発明に用いられるビフィドバクテリウム属細菌は、生菌であっても死菌であってもよく、生菌と死菌の両方を含むものでもよい。
　また、前記細菌の菌体処理物としては、例えば、菌体をアクリルアミドやカラギーナン等で固定化した固定化菌体、菌体の細胞壁及び細胞膜が超音波処理やホモジナイザー処理等の常法により一部又は完全に破砕された細胞破砕物等が挙げられる。
　さらに、前記細胞破砕物は前記破砕後の全画分でもよく一部の画分でもよく、前記破砕後の遠心分離上清、その上清を硫安処理等で部分精製した画分、又は前記上清を濃縮したものであってもよい。
　本発明に用いられるビフィドバクテリウム属細菌の具体的な形態としては、例えば、その菌体懸濁液、前記菌体（細菌）の細胞破砕物の水溶性画分、沈殿再懸濁液などが挙げられ、それらの中でも、非コラーゲン性糖タンパク質分解活性が高いことから、前記菌体（細菌）の細胞破砕物の水溶性画分が好ましい。 The Bifidobacterium genus bacterium used in the present invention can be used in the form of a bacterium itself, a culture of the bacterium, or a treated product of the bacterium as described above. That is, the bacterium belonging to the genus Bifidobacterium itself may be used, and after culturing the bacterium belonging to the genus Bifidobacterium, the obtained culture may be used as it is, or may be used after being diluted or concentrated. Moreover, the Bifidobacterium genus bacteria used for this invention may be live or dead, and may contain both live and dead.
Examples of the bacterial cell processed product include, for example, an immobilized bacterial cell in which the bacterial cell is fixed with acrylamide, carrageenan, or the like, and the cell wall and cell membrane of the bacterial cell are partially processed by an ordinary method such as ultrasonic treatment or homogenizer treatment. Or the completely crushed cell crushed material etc. are mentioned.
Further, the cell disrupted product may be the whole fraction after the disruption or a part of the fraction, the centrifuged supernatant after the disruption, the fraction obtained by partially purifying the supernatant by ammonium sulfate treatment, or the above The concentrate may be concentrated.
Specific examples of the Bifidobacterium genus bacteria used in the present invention include, for example, a cell suspension thereof, a water-soluble fraction of the cell disruption product of the cells (bacteria), and a resuspension suspension. Among them, the water-soluble fraction of the cell disruption product of the cells (bacteria) is preferable because of its high non-collagenous glycoprotein degradation activity.
　なお、本明細書における「菌体懸濁液」とは、本発明に用いられるビフィドバクテリウム属細菌の培養液を遠心分離等して得られる菌体を懸濁した溶液であり、好ましくは、後述する実施例の試験例３の（２）に記載される処理により得られる溶液である。
　また、本明細書における「水溶性画分」とは、上記細胞破砕物を遠心分離等して得られる上清であり、好ましくは、後述する実施例の試験例３の（２）に記載される処理により最終的に得られる上清である。該水溶性画分は、常法により適宜精製又は濃縮した水溶性画分がより好ましい。
　さらに、本明細書における「沈殿再懸濁液」とは、上記細胞破砕物を遠心分離等して得られる沈殿物を懸濁した溶液であり、好ましくは、後述する実施例の試験例３の（２）に記載される処理により最終的に得られる沈殿物を懸濁した溶液である。 The “bacterial cell suspension” in the present specification is a solution obtained by suspending a cell obtained by centrifuging a culture solution of the genus Bifidobacterium used in the present invention, preferably This is a solution obtained by the treatment described in (2) of Test Example 3 in Examples described later.
In addition, the “water-soluble fraction” in the present specification is a supernatant obtained by centrifuging the cell disrupted product, and is preferably described in (2) of Test Example 3 in Examples described later. It is a supernatant finally obtained by the process. The water-soluble fraction is more preferably a water-soluble fraction appropriately purified or concentrated by a conventional method.
Furthermore, the “precipitation resuspension” in the present specification is a solution in which a precipitate obtained by centrifuging the above-mentioned cell disruption is suspended, and preferably in Test Example 3 of Examples described later. It is a solution in which a precipitate finally obtained by the treatment described in (2) is suspended.
（２）非コラーゲン性糖タンパク質
　本発明の非コラーゲン性糖タンパク質としては、細胞や組織を囲む細胞外マトリックスの網目構造を形成する細胞接着分子が挙げられ、モノマーの状態でもポリマーの状態でもよい。具体的には、フィブリン（「安定化フィブリン」ともいう。）、フィブリン・ポリマー、フィブリン・モノマー、フィブリノゲン、フィブロネクチン、ラミニン、プラスミノーゲン、ビトロネクチン、ニドジェン、テネイシン、トロンボスポンジン、フォンビルブランド、オステオポンチン等を例示できる。これらの中でも、フィブリン、フィブリノゲン、フィブロネクチン、ラミニン、プラスミノーゲンが好ましく、フィブリノゲンがより好ましい。 (2) Non-collagenous glycoprotein The non-collagenous glycoprotein of the present invention includes cell adhesion molecules that form a network structure of an extracellular matrix surrounding cells and tissues, and may be in a monomer state or a polymer state. Specifically, fibrin (also referred to as “stabilized fibrin”), fibrin polymer, fibrin monomer, fibrinogen, fibronectin, laminin, plasminogen, vitronectin, nidogen, tenascin, thrombospondin, von Willebrand, osteopontin Etc. can be illustrated. Among these, fibrin, fibrinogen, fibronectin, laminin and plasminogen are preferable, and fibrinogen is more preferable.
＜医薬組成物＞
　本発明の非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物は、医薬組成物として用いることができる。
　プラスミンは、網膜剥離、網膜裂傷、硝子体出血、糖尿病性硝子体出血、増殖性糖尿、加齢黄斑変性、黄斑円孔、硝子体黄斑牽引、フィブリン沈着、網膜静脈閉塞症、網膜動脈閉塞症、緑内障及び網膜色素変性症等の眼の障害において、硝子体剥離が必要な化学的硝子体切除術；脳梗塞、四肢動静脈血栓症、肺梗塞及び脳静脈洞血栓等の虚血性障害に対する線溶療法に利用されている（特許文献２、非特許文献１）。このうち、脳梗塞、四肢動静脈血栓症、肺梗塞及び脳静脈洞血栓等の虚血性障害に対する線溶療法に有効であることは、プラスミンがフィブリノゲンの分解効果を有することに基づいている（非特許文献１）。
　したがって、本発明の医薬組成物は、網膜剥離、網膜裂傷、硝子体出血、糖尿病性硝子体出血、増殖性糖尿、加齢黄斑変性、黄斑円孔、硝子体黄斑牽引、フィブリン沈着、網膜静脈閉塞症、網膜動脈閉塞症、緑内障及び網膜色素変性症等の硝子体剥離が必要な眼病；脳梗塞、四肢動静脈血栓症、肺梗塞及び脳静脈洞血栓等の虚血性障害の予防及び／又は治療に使用することができる。
　また、本発明の医薬組成物は、経口組成物成分として長年使用されているビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とするため、種々の疾患を罹患した患者に対しても安心して投与できる。また、ビフィドバクテリウム属細菌は、動物の腸内にも存在するため、長期間、連続的に投与しても副作用が生じにくいことが期待される。また、本発明の非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とするため、乳幼児や小児にも安全に投与することができる。したがって、本発明の非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物は、乳幼児や小児の疾患の予防及び／又は治療に好適である。 <Pharmaceutical composition>
The non-collagenous glycoprotein degradation agent or non-collagenous glycoprotein degradation composition of the present invention can be used as a pharmaceutical composition.
Plasmin is a retinal detachment, retinal tear, vitreous hemorrhage, diabetic vitreous hemorrhage, proliferative diabetes, age-related macular degeneration, macular hole, vitreous macular traction, fibrin deposition, retinal vein occlusion, retinal artery occlusion, Chemical vitrectomy that requires vitreal detachment for glaucoma and retinitis pigmentosa; fibrinolysis for ischemic disorders such as cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction and cerebral venous sinus thrombus It is used for therapy (Patent Document 2, Non-Patent Document 1). Among these, plasmin has a fibrinogen decomposing effect because it is effective for fibrinolytic therapy for ischemic disorders such as cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction and cerebral venous sinus thrombus (non- Patent Document 1).
Therefore, the pharmaceutical composition of the present invention comprises retinal detachment, retinal laceration, vitreous hemorrhage, diabetic vitreous hemorrhage, proliferative diabetes, age-related macular degeneration, macular hole, vitreous macular traction, fibrin deposition, retinal vein occlusion Prevention and / or treatment of ischemic disorders such as cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction and cerebral venous sinus thrombosis Can be used for
Moreover, since the pharmaceutical composition of the present invention contains a Bifidobacterium genus bacterium, a culture of the bacterium, and / or a treated product of the bacterium that have been used for many years as an oral composition component, It can be administered with peace of mind to patients suffering from various diseases. In addition, since Bifidobacterium bacteria are also present in the intestines of animals, it is expected that side effects are unlikely to occur even when continuously administered for a long period of time. Further, the non-collagenous glycoprotein degrading agent or the non-collagenous glycoprotein degrading composition of the present invention comprises a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient. Therefore, it can be safely administered to infants and children. Therefore, the non-collagenous glycoprotein degrading agent or the non-collagenous glycoprotein degrading composition of the present invention is suitable for the prevention and / or treatment of diseases of infants and children.
　本発明に係る非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物を医薬組成物として利用する場合、該医薬組成物は、経口投与及び非経口投与のいずれでもよく、投与方法に応じて、適宜所望の剤形に製剤化することができる。例えば、経口投与の場合、散剤、顆粒剤、錠剤、カプセル剤等の固形製剤；溶液剤、シロップ剤、懸濁剤、乳剤等の液剤等に製剤化することができる。また、非経口投与の場合、座剤、軟膏剤又は点眼剤等に製剤化することができる。 When the non-collagenous glycoprotein degrading agent or the non-collagenous glycoprotein degrading composition according to the present invention is used as a pharmaceutical composition, the pharmaceutical composition may be administered either orally or parenterally. Accordingly, it can be appropriately formulated into a desired dosage form. For example, in the case of oral administration, it can be formulated into solid preparations such as powders, granules, tablets and capsules; liquid preparations such as solutions, syrups, suspensions and emulsions. In the case of parenteral administration, it can be formulated into a suppository, an ointment, an eye drop or the like.
　また、製剤化に際しては、本発明に係る非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物の他に、通常製剤化に用いられている賦形剤、ｐＨ調整剤、着色剤、矯味剤等の成分を用いることができる。また、本発明の効果を損なわない限り、本発明に係る非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物と、公知の又は将来的に見出される非コラーゲン性糖タンパク質が関与する疾患に対する予防及び／又は治療の効果を有する成分とを併用することもできる。 Further, in formulation, in addition to the non-collagenous glycoprotein degrading agent or non-collagenous glycoprotein degrading composition according to the present invention, excipients, pH adjusters, and colorants that are usually used for formulation. Ingredients such as flavoring agents can be used. In addition, as long as the effects of the present invention are not impaired, the non-collagenous glycoprotein degrading agent or the non-collagenous glycoprotein degrading composition according to the present invention and known or future non-collagenous glycoproteins are involved. A component having an effect of preventing and / or treating a disease can be used in combination.
　加えて、製剤化は剤形に応じて適宜公知の方法により実施できる。製剤化に際しては、適宜、製剤担体を配合して製剤化してもよい。 In addition, formulation can be performed by a known method as appropriate according to the dosage form. Upon formulation, a formulation carrier may be appropriately blended to formulate.
　本発明の医薬組成物の摂取量又は投与量は、剤形に合わせて適宜選択することができる。例えば、体重１ｋｇあたりの１日のビフィドバクテリウム属細菌の摂取量又は投与量は、１×１０６～１×１０１２ＣＦＵ／ｋｇ／日が好ましく、１×１０７～１×１０１１ＣＦＵ／ｋｇ／日がより好ましく、１×１０８～１×１０１０ＣＦＵ／ｋｇ／日がさらに好ましい。なお、前記単位のうちＣＦＵは、ｃｏｌｏｎｙ ｆｏｒｍｉｎｇ ｕｎｉｔｓの略であり、コロニー形成単位である。該細菌が死菌の場合、ＣＦＵは個細胞（cells）と置き換えることができる。
　また、ビフィドバクテリウム属細菌の培養物や前記細菌の菌体処理物を用いる場合のその摂取量又は投与量は、ビフィドバクテリウム属細菌の摂取量又は投与量に換算された場合に上記摂取量又は投与量となることが好ましい。 The intake or dose of the pharmaceutical composition of the present invention can be appropriately selected according to the dosage form. For example, the daily intake or dosage of Bifidobacterium per kg body weight is preferably 1 × 10 6 to 1 × 10 12 CFU / kg / day, and 1 × 10 7 to 1 × 10 11 CFU. / Kg / day is more preferable, and 1 × 10 8 to 1 × 10 10 CFU / kg / day is more preferable. In addition, CFU is an abbreviation of colony forming units and is a colony forming unit. If the bacterium is dead, the CFU can be replaced with cells.
In addition, when using a culture of Bifidobacterium bacteria or a treated product of the bacteria, the intake or dose is the above when converted to the intake or dose of Bifidobacterium. It is preferable to be an intake or a dose.
　また、本発明の医薬組成物中のビフィドバクテリウム属細菌の含有量は、前記摂取量又は投与量に基づいて適宜選択することができる。例えば、１×１０６～１×１０１２ＣＦＵ／ｇ又は１×１０６～１×１０１２ＣＦＵ／ｍＬ、好ましくは１×１０７～１×１０１１ＣＦＵ／ｇ又は１×１０７～１×１０１１ＣＦＵ／ｍＬ、より好ましくは１×１０８～１×１０１０ＣＦＵ／ｇ又は１×１０８～１×１０１０ＣＦＵ／ｍＬとすることができる。該細菌が死菌の場合、ＣＦＵは個細胞（cells）と置き換えることができる。
　また、ビフィドバクテリウム属細菌の培養物や前記細菌の菌体処理物を用いる場合のその含有量は、ビフィドバクテリウム属細菌の含有量に換算された場合に上記含有量となることが好ましい。 In addition, the content of Bifidobacterium in the pharmaceutical composition of the present invention can be appropriately selected based on the intake or dose. For example, 1 × 10 6 to 1 × 10 12 CFU / g or 1 × 10 6 to 1 × 10 12 CFU / mL, preferably 1 × 10 7 to 1 × 10 11 CFU / g or 1 × 10 7 to 1 × It can be 10 11 CFU / mL, more preferably 1 × 10 8 to 1 × 10 10 CFU / g or 1 × 10 8 to 1 × 10 10 CFU / mL. If the bacterium is dead, the CFU can be replaced with cells.
In addition, when using a culture of Bifidobacterium bacteria or a treated product of the bacteria, the content may be the above content when converted to the content of Bifidobacterium bacteria. preferable.
　また、前記製剤担体としては、剤形に応じて、各種有機又は無機の担体を用いることができる。固形製剤の場合の担体としては、例えば、賦形剤、結合剤、崩壊剤、滑沢剤、安定剤、矯味矯臭剤等が挙げられる。 In addition, as the preparation carrier, various organic or inorganic carriers can be used depending on the dosage form. Examples of the carrier in the case of a solid preparation include excipients, binders, disintegrants, lubricants, stabilizers, and flavoring agents.
　賦形剤としては、例えば、乳糖、白糖、ブドウ糖、マンニット、ソルビット等の糖誘導体；トウモロコシデンプン、馬鈴薯デンプン、α‐デンプン、デキストリン、カルボキシメチルデンプン等のデンプン誘導体；結晶セルロース、ヒドロキシプロピルセルロース、ヒドロキシプロピルメチルセルロース、カルボキシメチルセルロース、カルボキシメチルセルロースカルシウム等のセルロース誘導体；アラビアゴム；デキストラン；プルラン；軽質無水珪酸、合成珪酸アルミニウム、メタ珪酸アルミン酸マグネシウム等の珪酸塩誘導体；リン酸カルシウム等のリン酸塩誘導体；炭酸カルシウム等の炭酸塩誘導体；硫酸カルシウム等の硫酸塩誘導体等が挙げられる。 Examples of the excipient include sugar derivatives such as lactose, sucrose, glucose, mannitol and sorbit; starch derivatives such as corn starch, potato starch, α-starch, dextrin and carboxymethyl starch; crystalline cellulose, hydroxypropyl cellulose, Cellulose derivatives such as hydroxypropylmethylcellulose, carboxymethylcellulose, carboxymethylcellulose calcium; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, magnesium magnesium magnesium silicate; phosphate derivatives such as calcium phosphate; And carbonate derivatives such as calcium; sulfate derivatives such as calcium sulfate and the like.
　結合剤としては、例えば、上記賦形剤の他、ゼラチン；ポリビニルピロリドン；マクロゴール等が挙げられる。 Examples of the binder include gelatin, polyvinyl pyrrolidone, macrogol and the like in addition to the above excipients.
　崩壊剤としては、例えば、上記賦形剤の他、クロスカルメロースナトリウム、カルボキシメチルスターチナトリウム、架橋ポリビニルピロリドン等の化学修飾されたデンプン又はセルロース誘導体等が挙げられる。 Examples of the disintegrant include, in addition to the above excipients, chemically modified starch or cellulose derivatives such as croscarmellose sodium, sodium carboxymethyl starch, and crosslinked polyvinylpyrrolidone.
　滑沢剤としては、例えば、タルク；ステアリン酸；ステアリン酸カルシウム、ステアリン酸マグネシウム等のステアリン酸金属塩；コロイドシリカ；ピーガム、ゲイロウ等のワックス類；硼酸；グリコール；フマル酸、アジピン酸等のカルボン酸類；安息香酸ナトリウム等のカルボン酸ナトリウム塩；硫酸ナトリウム等の硫酸塩類；ロイシン；ラウリル硫酸ナトリウム、ラウリル硫酸マグネシウム等のラウリル硫酸塩；無水珪酸、珪酸水和物等の珪酸類；デンプン誘導体等が挙げられる。 As the lubricant, for example, talc; stearic acid; stearic acid metal salts such as calcium stearate and magnesium stearate; colloidal silica; waxes such as pea gum and geirow; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid Carboxylic acid sodium salts such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as anhydrous silicic acid and silicic acid hydrate; starch derivatives and the like It is done.
　安定剤としては、例えば、メチルパラベン、プロピルパラベン等のパラオキシ安息香酸エステル類；クロロブタノール、ベンジルアルコール、フェニルエチルアルコール等のアルコール類；塩化ベンザルコニウム；無水酢酸；ソルビン酸等が挙げられる。 Examples of the stabilizer include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; sorbic acid and the like.
　矯味矯臭剤としては、例えば、甘味料、酸味料、香料等が挙げられる。
　なお、経口投与用の液剤の場合に使用する担体としては、水等の溶剤、矯味矯臭剤等が挙げられる。 Examples of the flavoring agent include sweeteners, acidulants, and fragrances.
In addition, as a carrier used in the case of a liquid for oral administration, a solvent such as water, a flavoring agent and the like can be mentioned.
＜飲食品組成物＞
　さらに、本発明の非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物は、飲食品組成物として用いることができる。本発明の飲食品組成物は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を公知の飲食品に添加することによって製造してもよいし、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を飲食品の原料中に混合して新たな飲食品組成物として製造することもできる。また、本発明の飲食品組成物は、ビフィドバクテリウム属細菌を飲食品原料に添加した後に該ビフィドバクテリウム属細菌を培養する工程を含む製造方法により製造することもできる。
　ここで、プラスミンは、乳製品の熟成やチーズの製造にも利用されている（X. G. Song et al., 日本食品工業学会誌、第４０巻、第４号、１９９３年４月）。また、プラスミンは、哺乳動物の乳中にプラスミノーゲンとして存在しており、早産の母親の母乳は、通常よりもプラスミン濃度が高いことが知られている。その結果、早産児に与えられる母乳には、満期産児に与えられる母乳よりも高濃度の乳由来ペプチドが含まれている（E. Armaforte et al., International Dairy Journal, 20, pp.715-723, 2010）。
　したがって、本発明の非コラーゲン性糖タンパク質分解用飲食品組成物は、プラスミノーゲンを含むことが好ましく、乳製品やチーズの熟成促進のための添加用食品素材や、早産児向けの育児用調製粉乳、母乳又は食品への添加用食品素材として好適に用いることができる。 <Food and beverage composition>
Furthermore, the non-collagenous glycoprotein degrading agent or the non-collagenous glycoprotein degrading composition of the present invention can be used as a food or drink composition. The food / beverage composition of the present invention may be produced by adding a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium to a known food / beverage product. It is also possible to produce a new food / beverage product composition by mixing the bacteria belonging to the genus Fidobacterium, the culture of the bacteria, and / or the processed product of the bacteria into the raw material of the food / beverage product. Moreover, the food-drinks composition of this invention can also be manufactured by the manufacturing method including the process of culture | cultivating this Bifidobacterium genus bacteria, after adding a Bifidobacterium bacterium to a food-drinks raw material.
Here, plasmin is also used for ripening dairy products and producing cheese (X. G. Song et al., Journal of the Japan Food Industry Association, Vol. 40, No. 4, April 1993). Moreover, plasmin exists as plasminogen in mammalian milk, and it is known that the mother's milk of a premature mother has a higher plasmin concentration than usual. As a result, breast milk fed to preterm infants contains a higher concentration of milk-derived peptides than breast milk fed to full-term infants (E. Armaforte et al., International Dairy Journal, 20, pp.715-723 , 2010).
Accordingly, the non-collagenous glycoprotein degradation food / beverage composition of the present invention preferably contains plasminogen, an additive food material for promoting ripening of dairy products and cheese, and a preparation for childcare for premature infants. It can be suitably used as a food material for addition to powdered milk, breast milk or food.
　本発明における飲食品組成物は、液状、ペースト状、固体、粉末等の形態を問わず、錠菓、流動食、飼料（ペット用を含む）等のほか、例えば、小麦粉製品、即席食品、農産加工品、水産加工品、畜産加工品、乳・乳製品、油脂類、基礎調味料、複合調味料・食品類、冷凍食品、菓子類、飲料、これら以外の市販品等が挙げられる。また、本発明の効果を損なわない限り、本発明の飲食品組成物には、公知の又は将来的に見出されるプロバイオティクス効果を有する成分又はプロバイオティクス効果を補助する成分を使用することができる。例えば、本発明の飲食品組成物は、ホエイタンパク質、カゼインタンパク質、大豆タンパク質、若しくはエンドウ豆タンパク質（ピープロテイン）等の各種タンパク質若しくはその混合物、分解物；ロイシン、バリン、イソロイシン若しくはグルタミン等のアミノ酸；ビタミンＢ６若しくはビタミンＣ等のビタミン類；クレアチン；クエン酸；フィッシュオイル；又は、イソマルトオリゴ糖、ガラクトオリゴ糖、キシロオリゴ糖、大豆オリゴ糖、フラクトオリゴ糖、ラクチュロース等のオリゴ糖等の成分を含むことができる。 The food / beverage composition in the present invention may be in the form of liquid, paste, solid, powder, etc. In addition to tablet confectionery, liquid food, feed (including for pets), etc., for example, flour products, instant foods, agricultural products Processed products, processed fishery products, processed livestock products, milk / dairy products, fats and oils, basic seasonings, compound seasonings / foods, frozen foods, confectionery, beverages, commercial products other than these. In addition, as long as the effects of the present invention are not impaired, the food / beverage composition of the present invention may use a component having a probiotic effect known in the future or a component assisting the probiotic effect. it can. For example, the food / beverage composition of the present invention comprises various proteins such as whey protein, casein protein, soy protein, pea protein (pea protein) or mixtures thereof, and degradation products thereof; amino acids such as leucine, valine, isoleucine or glutamine; Vitamins such as vitamin B6 or vitamin C; creatine; citric acid; fish oil; or components such as isomalt-oligosaccharide, galactooligosaccharide, xylo-oligosaccharide, soybean oligosaccharide, fructooligosaccharide, lactulose .
　また、本発明で定義される飲食品組成物は、非コラーゲン性糖タンパク質が有効に作用する疾患の予防、疾患のリスク低減、疾患の症状緩和、及び／又は治療等の用途（保健用途を含む）が表示された飲食品として提供・販売されることが可能である。
　「表示」行為には、需要者に対して前記用途を知らしめるための全ての行為が含まれ、前記用途を想起・類推させうるような表現であれば、表示の目的、表示の内容、表示する対象物・媒体等の如何に拘わらず、全て本発明の「表示」行為に該当する。 In addition, the food / beverage product composition defined in the present invention is used for prevention of diseases in which non-collagenous glycoproteins effectively act, risk reduction of diseases, symptom alleviation of diseases, and / or treatment (including health uses). ) Can be provided and sold as a food or drink.
The “display” act includes all acts for informing the consumer of the use, and if the expression can remind the user of the use, the purpose of the display, the content of the display, the display Regardless of the target object / medium, etc., all fall under the “display” act of the present invention.
　また、「表示」は、需要者が上記用途を直接的に認識できるような表現により行われることが好ましい。具体的には、飲食品に係る商品又は商品の包装に前記用途を記載したものを譲渡し、引き渡し、譲渡若しくは引き渡しのために展示し、輸入する行為、商品に関する広告、価格表若しくは取引書類に上記用途を記載して展示し、若しくは頒布し、又はこれらを内容とする情報に上記用途を記載して電磁気的（インターネット等）方法により提供する行為等が挙げられる。 In addition, it is preferable that the “display” is performed by an expression that allows the consumer to directly recognize the use. Specifically, it is the act of transferring, displaying, importing, displaying, or importing products that are related to food or drinks or products that describe the use, on advertisements, price lists, or transaction documents. For example, an act of describing and displaying the above uses or distributing them, or describing the above uses in information including the contents and providing them by an electromagnetic (Internet or the like) method can be given.
　一方、表示内容としては、行政等によって認可された表示（例えば、行政が定める各種制度に基づいて認可を受け、そのような認可に基づいた態様で行う表示等）であることが好ましい。また、そのような表示内容を、包装、容器、カタログ、パンフレット、ＰＯＰ等の販売現場における宣伝材、その他の書類等へ付することが好ましい。 On the other hand, the display content is preferably a display approved by the government or the like (for example, a display that is approved based on various systems determined by the government and is performed in a mode based on such approval). Moreover, it is preferable to attach such display contents to advertising materials at sales sites such as packaging, containers, catalogs, pamphlets, POPs, and other documents.
　また、「表示」には、健康食品、機能性食品、経腸栄養食品、特別用途食品、保健機能食品、特定保健用食品、栄養機能食品、機能性表示食品、医薬用部外品等としての表示も挙げられる。この中でも特に、消費者庁によって認可される表示、例えば、特定保健用食品、栄養機能食品、若しくは機能性表示食品に係る制度、又はこれらに類似する制度にて認可される表示等が挙げられる。具体的には、特定保健用食品としての表示、条件付き特定保健用食品としての表示、身体の構造や機能に影響を与える旨の表示、疾病リスク減少表示、科学的根拠に基づいた機能性の表示等を挙げることができ、より具体的には、健康増進法に規定する特別用途表示の許可等に関する内閣府令（平成二十一年八月三十一日内閣府令第五十七号）に定められた特定保健用食品としての表示（特に保健の用途の表示）及びこれに類する表示が典型的な例である。 In addition, “labeling” includes health food, functional food, enteral nutrition food, special purpose food, health functional food, food for specified health use, nutrition functional food, functional label food, quasi-drug, etc. A display is also included. Among these, in particular, indications approved by the Consumer Affairs Agency, for example, indications approved in systems related to foods for specified health use, functional nutritional foods, functional indication foods, or similar systems, etc. can be mentioned. Specifically, labeling as a food for specified health use, labeling as a conditionally specified food for specified health use, labeling that affects the structure and function of the body, labeling for reducing the risk of disease, and functionality based on scientific evidence Labeling, etc., and more specifically, Cabinet Office Ordinance concerning permission for special purpose labeling provided for in the Health Promotion Act (Cabinet Office Ordinance No. 57, August 31, 2000) The labeling as food for specified health (particularly the labeling of health use) and the like are the typical examples.
　本発明の飲食品組成物の摂取量は、適宜選択することができる。例えば、体重１ｋｇあたりの１日のビフィドバクテリウム属細菌の摂取量として、１×１０６～１×１０１２ＣＦＵ／ｋｇ／日が好ましく、１×１０７～１×１０１１ＣＦＵ／ｋｇ／日がより好ましく、１×１０８～１×１０１０ＣＦＵ／ｋｇ／日がさらに好ましい。該細菌が死菌の場合、ＣＦＵは個細胞（cells）と置き換えることができる。
　また、ビフィドバクテリウム属細菌の培養物や前記細菌の菌体処理物を用いる場合のその摂取量は、ビフィドバクテリウム属細菌の摂取量に換算された場合に上記摂取量となることが好ましい。 The intake of the food / beverage product composition of the present invention can be appropriately selected. For example, the daily intake of Bifidobacterium per kg of body weight is preferably 1 × 10 6 to 1 × 10 12 CFU / kg / day, and 1 × 10 7 to 1 × 10 11 CFU / kg / day. Day is more preferable, and 1 × 10 8 to 1 × 10 10 CFU / kg / day is more preferable. If the bacterium is dead, the CFU can be replaced with cells.
In addition, when using a culture of Bifidobacterium bacteria or a treated product of the bacteria, the intake may be the above intake when converted to the intake of Bifidobacterium. preferable.
　また、本発明の飲食品組成物中のビフィドバクテリウム属細菌の含有量は、前記摂取量に基づいて適宜選択することができる。例えば、１×１０６～１×１０１２ＣＦＵ／ｇ又は１×１０６～１×１０１２ＣＦＵ／ｍＬ、好ましくは１×１０７～１×１０１１ＣＦＵ／ｇ又は１×１０７～１×１０１１ＣＦＵ／ｍＬ、より好ましくは１×１０８～１×１０１０ＣＦＵ／ｇ又は１×１０８～１×１０１０ＣＦＵ／ｍＬとすることができる。該細菌が死菌の場合、ＣＦＵは個細胞（cells）と置き換えることができる。
　また、ビフィドバクテリウム属細菌の培養物や前記細菌の菌体処理物を用いる場合のその含有量は、ビフィドバクテリウム属細菌の含有量に換算された場合に上記含有量となることが好ましい。 In addition, the content of Bifidobacterium in the food and beverage composition of the present invention can be appropriately selected based on the intake. For example, 1 × 10 6 to 1 × 10 12 CFU / g or 1 × 10 6 to 1 × 10 12 CFU / mL, preferably 1 × 10 7 to 1 × 10 11 CFU / g or 1 × 10 7 to 1 × It can be 10 11 CFU / mL, more preferably 1 × 10 8 to 1 × 10 10 CFU / g or 1 × 10 8 to 1 × 10 10 CFU / mL. If the bacterium is dead, the CFU can be replaced with cells.
In addition, when using a culture of Bifidobacterium bacteria or a treated product of the bacteria, the content may be the above content when converted to the content of Bifidobacterium bacteria. preferable.
　本発明に係る非コラーゲン性糖タンパク質分解用飲食品組成物は、ヒト若しくは動物用の飲食品組成物として使用することができる。また、本発明に係る非コラーゲン性糖タンパク質分解用飲食品組成物は、脳梗塞、四肢動静脈血栓症、肺梗塞、脳静脈洞血栓、網膜剥離、網膜裂傷、硝子体出血、糖尿病性硝子体出血、増殖性糖尿、加齢黄斑変性、黄斑円孔、硝子体黄斑牽引、フィブリン沈着、網膜静脈閉塞症、網膜動脈閉塞症、緑内障又は網膜色素変性症等の非コラーゲン性糖タンパク質が関連する疾患の予防及び／又は治療に有効である。本発明の非コラーゲン性糖タンパク質分解用飲食品組成物は、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とするため、乳幼児や小児にも安全に投与することができる。したがって、本発明の非コラーゲン性糖タンパク質分解用飲食品組成物は、乳幼児や小児における疾患の予防及び／又は治療にも好適である。 The non-collagenous glycoprotein degrading food / beverage composition according to the present invention can be used as a human or animal food / beverage composition. In addition, the non-collagenous glycoprotein degradation food / beverage composition according to the present invention includes cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction, cerebral venous sinus thrombus, retinal detachment, retinal tear, vitreous hemorrhage, diabetic vitreous Non-collagenous glycoprotein-related diseases such as bleeding, proliferative diabetes, age-related macular degeneration, macular hole, vitreous macular traction, fibrin deposition, retinal vein occlusion, retinal artery occlusion, glaucoma or retinitis pigmentosa It is effective for prevention and / or treatment. The food and drink composition for degrading non-collagenous glycoprotein of the present invention contains Bifidobacterium, culture of the bacterium, and / or treated product of the bacterium as an active ingredient. Can also be safely administered. Therefore, the non-collagenous glycoprotein degradation food / beverage composition of the present invention is also suitable for the prevention and / or treatment of diseases in infants and children.
＜プラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質の製造方法＞
　本発明の他の実施態様は、プラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質の製造方法である。本実施態様は、ビフィドバクテリウム属細菌を培養する工程（培養工程）、及び前記培養後の培養物中に産生されたプラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質を回収する工程（回収工程）を含むが、その他の工程を含んでもよい。 <Method for producing protein having plasminogen activator activity and / or plasmin activity>
Another embodiment of the present invention is a method for producing a protein having plasminogen activator activity and / or plasmin activity. In the present embodiment, a step of culturing Bifidobacterium (culture step), and a step of recovering a protein having plasminogen activator activity and / or plasmin activity produced in the culture after the culture (Recovery step) is included, but other steps may be included.
　プラスミノーゲンアクチベーター活性を有するタンパク質とプラスミン活性を有するタンパク質は、一のタンパク質でもあってもよく、異なるタンパク質であってもよい。すなわち、一のタンパク質としてプラスミノーゲンアクチベーター活性とプラスミン活性の両方を有していてもよいし、プラスミノーゲンアクチベーター活性を有するタンパク質とプラスミン活性を有するタンパク質として別個のタンパク質であってもよい。 The protein having plasminogen activator activity and the protein having plasmin activity may be one protein or different proteins. That is, one protein may have both plasminogen activator activity and plasmin activity, or a protein having plasminogen activator activity and a protein having plasmin activity may be separate proteins. .
（培養工程）
　本工程はビフィドバクテリウム属細菌を培養する工程であり、ビフィドバクテリウム属細菌を培養できる公知の培養条件を採用することができる。例えば、２５～４５℃で培養することが可能であるが、３０～４２℃で培養することが好ましく、３７～４２℃で培養することがより好ましい。また、培養は嫌気条件下で行うことが好ましく、例えば、炭酸ガス等の嫌気ガスを通気しながら培養することができる。また、液体静置培養等の微好気条件下で培養することも可能である。培養時間は、１～７２時間の間で増殖速度を観察しながら適宜調節可能であるが、１６～４８時間が好ましく、１６～２４時間がより好ましい。 (Culture process)
This step is a step of culturing Bifidobacterium bacteria, and known culture conditions that can culture Bifidobacterium bacteria can be employed. For example, it can be cultured at 25 to 45 ° C., but it is preferably cultured at 30 to 42 ° C., more preferably 37 to 42 ° C. Moreover, it is preferable to perform culture | cultivation on anaerobic conditions, for example, it can culture | cultivate, ventilating anaerobic gas, such as a carbon dioxide gas. It is also possible to culture under microaerobic conditions such as liquid stationary culture. The culture time can be appropriately adjusted while observing the growth rate between 1 and 72 hours, but is preferably 16 to 48 hours, more preferably 16 to 24 hours.
（回収工程）
　本工程は、前記培養後の培養物中に産生されたプラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質を回収する工程である。本工程は、（ａ）前記培養後の培養物を、ビフィドバクテリウム属細菌とプラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質を含む画分とに分離する工程と、（ｂ）該画分からプラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質を回収する工程を含むが、その他の工程を含んでもよい。工程（ａ）と（ｂ）とは同時に行われてもよく、工程（ａ）後に工程（ｂ）が行われてもよい。 (Recovery process)
This step is a step of recovering a protein having plasminogen activator activity and / or plasmin activity produced in the culture after the culture. This step includes (a) separating the culture after the culture into a Bifidobacterium genus and a fraction containing a protein having plasminogen activator activity and / or plasmin activity, and (b) The method includes a step of recovering a protein having plasminogen activator activity and / or plasmin activity from the fraction, but may include other steps. Steps (a) and (b) may be performed simultaneously, and step (b) may be performed after step (a).
　工程（ａ）では、公知の方法を採用することができ、例えば、膜によるろ過、遠心分離等の方法を採用することができる。該膜は、平膜及び中空糸膜（ホローファイバー）のいずれでもよい。 In the step (a), a known method can be employed, and for example, a method such as filtration with a membrane or centrifugation can be employed. The membrane may be either a flat membrane or a hollow fiber membrane (hollow fiber).
　また、工程（ａ）は、ビフィドバクテリウム属細菌の菌体を処理する工程を含んでもよい。すなわち、工程（ａ）は、ビフィドバクテリウム属細菌の菌体を処理する工程と、該菌体処理物とプラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質を含む画分とに分離する工程とを含んでもよい。ビフィドバクテリウム属細菌の菌体を処理する工程については、既に説明した、菌体処理物に関する記載が援用される。 Further, the step (a) may include a step of treating cells of the genus Bifidobacterium. That is, the step (a) is separated into a step of treating cells of the genus Bifidobacterium and a fraction containing the treated product and a protein having plasminogen activator activity and / or plasmin activity. And a step of performing. About the process of processing the microbial cell of Bifidobacterium genus, the description regarding the microbial cell processed material already demonstrated is used.
　工程（ｂ）では、公知の方法を採用することができ、例えば、ゲルろ過クロマトグラフィーや逆相ＨＰＬＣ等の各種クロマトグラフィー等の方法が挙げられる。クロマトグラフィーは、低圧であっても高圧であってもよい。
　中空糸膜（ホローファイバー）を使用してろ過を行った場合、前記工程（ａ）と（ｂ）とを同時に行うことができる。 In the step (b), a known method can be employed, and examples thereof include various chromatography methods such as gel filtration chromatography and reverse phase HPLC. Chromatography may be low or high pressure.
When filtration is performed using a hollow fiber membrane (hollow fiber), the steps (a) and (b) can be performed simultaneously.
　プラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質を含む画分は、プラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質の効果を奏する限り、培地成分等の他の成分を含んでいてもよく、該成分等を完全に又は部分的に精製したものであってもよく、その態様は特に制限されない。その精製は、上記工程（ａ）及び／又は（ｂ）における方法を適宜組み合わせることによって行うことができる。
　プラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質を含む画分の性状は、液体であってもよく、凍結乾燥等によって得られる粉体であってもよく、その態様は特に制限されない。 The fraction containing a protein having plasminogen activator activity and / or plasmin activity contains other components such as medium components as long as the effect of the protein having plasminogen activator activity and / or plasmin activity is exhibited. The component or the like may be completely or partially purified, and the mode is not particularly limited. The refinement | purification can be performed by combining suitably the method in the said process (a) and / or (b).
The properties of the fraction containing a protein having plasminogen activator activity and / or plasmin activity may be liquid or powder obtained by lyophilization or the like, and the mode is not particularly limited.
　本実施態様により製造されたプラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質は、これらが有する生理作用に基づいて、医薬、医薬部外品、皮膚外用剤、化粧料、飲食品、食品添加剤及び飼料等の組成物に配合することができる。
　また、上記の通り、哺乳動物の乳中にはプラスミノーゲンが存在することから、本実施態様により製造されたプラスミノーゲンアクチベーター活性を有するタンパク質を、乳製品やチーズの熟成促進のための添加用食品素材や、早産児向けの育児用調製粉乳、母乳又は食品への添加用食品素材として好適に用いることができる。 The protein having plasminogen activator activity and / or plasmin activity produced according to the present embodiment is based on the physiological action of the protein, quasi-drug, external preparation for skin, cosmetics, food and drink, food. It can mix | blend with compositions, such as an additive and feed.
Further, as described above, since plasminogen is present in the milk of mammals, the protein having plasminogen activator activity produced according to this embodiment is used to promote ripening of dairy products and cheese. It can be suitably used as a food material for additives, a formula powder for childcare for preterm infants, breast milk or food materials for addition to foods.
＜プラスミンの製造方法＞
　本発明の他の実施態様はプラスミンの製造方法である。本実施態様は、プラスミノーゲンを含む培地中でビフィドバクテリウム属細菌を培養する工程（培養工程）、及び前記培養後の培養物中に産生されたプラスミンを回収する工程（回収工程）を含むが、その他の工程を含んでもよい。 <Method for producing plasmin>
Another embodiment of the present invention is a method for producing plasmin. This embodiment includes a step of culturing Bifidobacterium in a medium containing plasminogen (culturing step), and a step of recovering plasmin produced in the culture after the culturing (recovering step). However, other steps may be included.
（培養工程）
　本工程は、プラスミノーゲンを含む培地中でビフィドバクテリウム属細菌を培養する工程である。培地へプラスミノーゲンを添加する場合には、培養前に添加しても、培養中に添加してもよい。また、一括添加、逐次添加、連続添加のいずれであってもよい。培地中のプラスミノーゲンの含有量は特に制限されない。
　ビフィドバクテリウム属細菌の培養条件等には、前記＜プラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質の製造方法＞の「培養工程」の記載が援用される。 (Culture process)
This step is a step of culturing Bifidobacterium in a medium containing plasminogen. When plasminogen is added to the medium, it may be added before culturing or during culturing. Moreover, any of collective addition, sequential addition, and continuous addition may be sufficient. The content of plasminogen in the medium is not particularly limited.
The description of the “culturing step” in <Method for producing protein having plasminogen activator activity and / or plasmin activity> is incorporated in the culture conditions of Bifidobacterium.
（回収工程）
　本工程は、前記培養後の培養物中に産生されたプラスミンを回収する工程である。本工程は、（ｃ）前記培養後の培養物を、ビフィドバクテリウム属細菌とプラスミンを含む画分とに分離する工程と、（ｄ）該画分からプラスミンを回収する工程を含むが、その他の工程を含んでもよい。工程（ｃ）と（ｄ）とは同時に行われてもよく、工程（ｃ）後に工程（ｄ）が行われてもよい。
　また、工程（ｃ）は、ビフィドバクテリウム属細菌の菌体を処理する工程を含んでもよい。すなわち、工程（ｃ）は、ビフィドバクテリウム属細菌の菌体を処理する工程と、該菌体処理物とプラスミンを含む画分とに分離する工程とを含んでもよい。ビフィドバクテリウム属細菌の菌体を処理する工程については、既に説明した、菌体処理物に関する記載が援用される。
　回収の方法や条件には、前記＜プラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質の製造方法＞の「回収工程」の記載が援用される。 (Recovery process)
This step is a step of recovering plasmin produced in the culture after the culture. This step includes (c) a step of separating the culture after the culture into a fraction containing Bifidobacterium and plasmin, and (d) a step of recovering plasmin from the fraction. These steps may be included. Steps (c) and (d) may be performed simultaneously, and step (d) may be performed after step (c).
Further, the step (c) may include a step of treating cells of the genus Bifidobacterium. That is, the step (c) may include a step of treating the cells of the genus Bifidobacterium, and a step of separating the treated cells and a fraction containing plasmin. About the process of processing the microbial cell of Bifidobacterium genus, the description regarding the microbial cell processed material already demonstrated is used.
The description of “recovery step” in the above <Method for producing protein having plasminogen activator activity and / or plasmin activity> is incorporated into the recovery method and conditions.
　本実施態様により製造されたプラスミンは、これが有する生理作用に基づいて、医薬、医薬部外品、皮膚外用剤、化粧料、飲食品、食品添加剤及び飼料等の組成物に配合することができる。また、本実施態様により製造されたプラスミンを、乳製品やチーズへの添加用食品素材や、早産児向けの育児用調製粉乳、母乳又は食品への添加用食品素材として好適に用いることができる。 The plasmin produced according to this embodiment can be blended in compositions such as pharmaceuticals, quasi drugs, external preparations for skin, cosmetics, foods and drinks, food additives, and feeds based on the physiological functions of the plasmin. . Moreover, the plasmin produced by this embodiment can be suitably used as a food material for addition to dairy products and cheese, a formula powder for childcare for premature infants, breast milk or a food material for addition to food.
　さらに、本発明は、以下の構成を採用することも可能である。尚、下記の哺乳動物としては、ヒト、ウシ、ヒツジ、ヤギ、ブタ、イヌ、ネコ、ウマ等が挙げられる。
［１］非コラーゲン性糖タンパク質分解用組成物又は非コラーゲン性糖タンパク質分解用医薬の製造における、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物の使用。
［２］非コラーゲン性糖タンパク質分解に用いられるビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物。
［３］非コラーゲン性糖タンパク質の分解によって緩和、予防又は治療され得る疾患の緩和、予防又は治療に用いられるビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物。
［４］非コラーゲン性糖タンパク質分解のためのビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物の使用。
［５］ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を哺乳動物に投与する段階を含む、非コラーゲン性糖タンパク質を分解する方法。
［６］ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とする非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物を哺乳動物に投与する段階を含む、非コラーゲン性糖タンパク質を分解する方法。
［７］ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を哺乳動物に投与する段階を含む、非コラーゲン性糖タンパク質の分解によって緩和、予防又は治療され得る疾患の緩和方法、予防方法又は治療方法。
［８］ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とする非コラーゲン性糖タンパク質分解剤又は非コラーゲン性糖タンパク質分解用組成物を哺乳動物に投与する段階を含む、非コラーゲン性糖タンパク質の分解によって緩和、予防又は治療され得る疾患の緩和方法、予防方法又は治療方法。 Furthermore, the present invention can employ the following configurations. The following mammals include humans, cows, sheep, goats, pigs, dogs, cats, horses and the like.
[1] Use of a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium in the manufacture of a composition for degrading non-collagenous glycoprotein or a medicament for degrading non-collagenous glycoprotein .
[2] Bifidobacterium used for non-collagenous glycoprotein degradation, culture of the bacterium, and / or treated product of the bacterium.
[3] Bifidobacterium genus bacteria used for the alleviation, prevention or treatment of diseases that can be alleviated, prevented or treated by degradation of non-collagenous glycoprotein, culture of the bacteria, and / or cell treatment of the bacteria object.
[4] Use of Bifidobacterium, a culture of the bacterium, and / or a treated product of the bacterium for non-collagenous glycoprotein degradation.
[5] A method for degrading non-collagenous glycoprotein, comprising administering a Bifidobacterium genus bacterium, a culture of the bacterium, and / or a treated product of the bacterium to a mammal.
[6] A non-collagenous glycoprotein degrading agent or a composition for degrading non-collagenous glycoprotein comprising a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient A method for degrading non-collagenous glycoproteins comprising the step of administering to an animal.
[7] It is alleviated, prevented or treated by degradation of non-collagenous glycoprotein, comprising administering a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium to a mammal. A method for alleviating, preventing or treating a disease obtained.
[8] A non-collagenous glycoprotein degrading agent or a composition for degrading non-collagenous glycoprotein comprising a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient A method for alleviating, preventing or treating a disease that can be alleviated, prevented or treated by degradation of non-collagenous glycoprotein, comprising a step of administering to an animal.
　以下に実施例を用いて本発明を説明するが、本発明はこれら実施例に限定されるものではない。 Hereinafter, the present invention will be described using examples, but the present invention is not limited to these examples.
［試験例１］
　ビフィドバクテリウム属細菌が、プラスミノーゲンアクチベーター活性を有することを確認する試験を行った。 [Test Example 1]
A test was conducted to confirm that Bifidobacterium has plasminogen activator activity.
（１）培養液の調製
　グルタミン酸ナトリウム１％及び脱脂粉乳１０％を含む水溶液にて凍結保存された以下の１７種のビフィドバクテリウム属細菌（ＨＲＢ１２種及びｎｏｎ‐ＨＲＢ５種）の菌液１００μＬを、それぞれＭＲＳ液体培地３ｍＬに添加し、ビフィドバクテリウム属細菌の菌数が１×１０９ＣＦＵ／ｍＬとなるように、３７℃で１６時間嫌気培養した。ＭＲＳ液体培地は、Difco Lactobacilli MRS Broth（BD社製）５．５ｇ、及びL-Cysteine Monohydrochloride, Monohydrate（和光純薬工業社製）５０ｍｇを、１００ｍＬとなるように純水に溶解させ、ＨＣｌ水溶液でｐＨ６．５に調整し、１２１℃で１５分間滅菌することによって調製した。
＜ヒト常在性のビフィドバクテリウム属細菌（ＨＲＢ）＞
・Bifidobacterium longum subsp. longum ATCC 15707
・Bifidobacterium longum subsp. longum ATCC BAA-999
・Bifidobacterium longum subsp. infantis ATCC 15697
・Bifidobacterium longum subsp. infantis BCCM LMG23728
・Bifidobacterium breve ATCC 15700
・Bifidobacterium breve FERM BP-11175
・Bifidobacterium breve BCCM LMG23729
・Bifidobacterium bifidum ATCC 29521
・Bifidobacterium adolescentis ATCC 15703
・Bifidobacterium angulatum ATCC 27535
・Bifidobacterium dentium DSM 20436
・Bifidobacterium psudocatenulatum ATCC 27919
＜非ヒト常在性のビフィドバクテリウム属細菌（ｎｏｎ‐ＨＲＢ）＞
・Bifidobacterium animalis subsp. lactis DSM 10140
・Bifidobacterium animalis subsp. animalis ATCC 25527
・Bifidobacterium pseudolongum subsp. globosum JCM 5820
・Bifidobacterium pseudolongum subsp. pseudolongm ATCC 25526
・Bifidobacterium thermophilum ATCC 25525 (1) Preparation of culture solution 100 μL of the following 17 kinds of Bifidobacterium bacteria (HRB12 and non-HRB5) frozen and stored in an aqueous solution containing 1% sodium glutamate and 10% nonfat dry milk Each was added to 3 mL of MRS liquid medium and anaerobically cultured at 37 ° C. for 16 hours so that the number of Bifidobacterium was 1 × 10 9 CFU / mL. The MRS liquid medium is prepared by dissolving 5.5 g of Difco Lactobacilli MRS Broth (BD) and 50 mg of L-Cysteine Monohydrochloride, Monohydrate (Wako Pure Chemical Industries) in pure water so as to be 100 mL, and using an aqueous HCl solution. It was prepared by adjusting to pH 6.5 and sterilizing at 121 ° C. for 15 minutes.
<Human resident Bifidobacterium (HRB)>
・ Bifidobacterium longum subsp. Longum ATCC 15707
・ Bifidobacterium longum subsp. Longum ATCC BAA-999
・ Bifidobacterium longum subsp. Infantis ATCC 15697
・ Bifidobacterium longum subsp. Infantis BCCM LMG23728
・ Bifidobacterium breve ATCC 15700
・ Bifidobacterium breve FERM BP-11175
・ Bifidobacterium breve BCCM LMG23729
・ Bifidobacterium bifidum ATCC 29521
・ Bifidobacterium adolescentis ATCC 15703
・ Bifidobacterium angulatum ATCC 27535
・ Bifidobacterium dentium DSM 20436
Bifidobacterium psudocatenulatum ATCC 27919
<Non-human resident Bifidobacterium (non-HRB)>
・ Bifidobacterium animalis subsp. Lactis DSM 10140
・ Bifidobacterium animalis subsp.animalis ATCC 25527
・ Bifidobacterium pseudolongum subsp.globosum JCM 5820
・ Bifidobacterium pseudolongum subsp.pseudolongm ATCC 25526
・ Bifidobacterium thermophilum ATCC 25525
（２）プラスミノーゲンアクチベーター活性の測定
　（１）にて調製した各培養液を４℃、５０００×ｇの条件下にて３０分間遠心処理した後、上清を捨て、分離した菌体をＰＢＳ溶液に懸濁した。各懸濁液は、濁度（ＯＤ６００）を０．１に揃えて調製し、プラスミノーゲンアクチベーターに特異的な蛍光基質であるＺ‐Ｇｌｙ‐Ｇｌｙ‐Ａｒｇ‐ＡＭＣ・ＨＣｌ（ＢＡＣＨＥＭ社製）を添加して６０分間、３７℃で嫌気的に培養した。培養終了後、ｍｉｃｒｏｐｌａｔｅ　ｒｅａｄｅｒ　ＳＨ‐９０００（Corona Electric社製）を用いて励起波長３６０ｎｍ、測定波長４６０ｎｍにて培養物の蛍光強度を測定した。なお、蛍光強度の単位は、任意単位（arbitrary unit:a.u.）とした。測定した蛍光強度から、あらかじめ作成した前記蛍光基質由来の蛍光物質（ＡＭＣ：アミノメチルクマリン）濃度と測定波長４６０ｎｍにおける蛍光強度との関係を示す検量線を用いて、培養物中のＡＭＣ濃度を算出し、当該濃度から、１分間に産生されるＡＭＣの１ｎｍｏｌを酵素１単位（Ｕ）として、各ビフィドバクテリウム属細菌の酵素活性（プラスミノーゲンアクチベーター活性）を算出した。 (2) Measurement of plasminogen activator activity Each culture solution prepared in (1) was centrifuged at 4 ° C. and 5000 × g for 30 minutes, the supernatant was discarded, and the separated cells were Suspended in PBS solution. Each suspension was prepared with a turbidity (OD600) of 0.1 and Z-Gly-Gly-Arg-AMC.HCl (manufactured by BACHEM), which is a fluorescent substrate specific for plasminogen activator. And anaerobically cultured at 37 ° C. for 60 minutes. After completion of the culture, the fluorescence intensity of the culture was measured using a microplate reader SH-9000 (manufactured by Corona Electric) at an excitation wavelength of 360 nm and a measurement wavelength of 460 nm. The unit of fluorescence intensity was an arbitrary unit (au). From the measured fluorescence intensity, the AMC concentration in the culture is calculated using a calibration curve showing the relationship between the fluorescent substance-derived fluorescent substance (AMC: aminomethylcoumarin) concentration prepared in advance and the fluorescence intensity at a measurement wavelength of 460 nm. From this concentration, the enzyme activity (plasminogen activator activity) of each Bifidobacterium was calculated using 1 nmol of AMC produced per minute as 1 unit of enzyme (U).
（３）結果
　各ビフィドバクテリウム属細菌の蛍光強度及び酵素活性（プラスミノーゲンアクチベーター活性）は、表１のようになり、全てのビフィドバクテリウム属細菌が、プラスミノーゲンアクチベーター活性を示すことが確認された。中でもBifidobacterium bifidum ATCC29521、Bifidobacterium breve FERM BP-11175及びBifidobacterium longum subsp. infantis ATCC15697等の乳幼児由来のビフィドバクテリウム属細菌のプラスミノーゲンアクチベーター活性は、ＨＲＢの中でもとりわけ高く、ｎｏｎ‐ＨＲＢのBifidobacterium animalis subsp. lactis DSM10140のプラスミノーゲンアクチベーター活性と比べると、いずれも１０倍以上を示し、強いプラスミノーゲンアクチベーター活性を有することが示唆された。 (3) Results The fluorescence intensity and enzyme activity (plasminogen activator activity) of each Bifidobacterium genus are as shown in Table 1, and all Bifidobacterium bacteria have plasminogen activator activity. It was confirmed that Among them, the plasminogen activator activity of Bifidobacterium bacteria such as Bifidobacterium bifidum ATCC29521, Bifidobacterium breve FERM BP-11175 and Bifidobacterium longum subsp. Infantis ATCC15697 is particularly high among HRB, and non-HRB Bifidobacterium animalis Compared with the plasminogen activator activity of subsp. lactis DSM10140, all showed 10 times or more, suggesting that it has strong plasminogen activator activity.
Figure JPOXMLDOC01-appb-T000001
Figure JPOXMLDOC01-appb-T000001
［試験例２］
　次に、ビフィドバクテリウム属細菌が、プラスミン活性を有することを確認する試験を行った。 [Test Example 2]
Next, a test was conducted to confirm that Bifidobacterium bacteria have plasmin activity.
（１）培養液の調製
　試験例１の（１）と同様の手順で、前記１７種のビフィドバクテリウム属細菌の培養液を調製した。 (1) Preparation of culture solution By the same procedure as in Test Example 1 (1), culture solutions of the 17 species of Bifidobacterium were prepared.
（２）プラスミン活性の測定
　プラスミノーゲンアクチベーターに特異的な蛍光基質ではなく、プラスミンに特異的な蛍光基質であるＢｏｃ‐Ｖａｌ‐Ｌｅｕ‐Ｌｙｓ‐ＡＭＣ酢酸塩（ＢＡＣＨＥＭ社製）を用いたこと以外は上記試験例１と同様にして、各ビフィドバクテリウム属細菌の酵素活性（プラスミン活性）を算出した。 (2) Measurement of plasmin activity Boc-Val-Leu-Lys-AMC acetate (manufactured by BACHEM), a fluorescent substrate specific for plasmin, was used instead of a fluorescent substrate specific for plasminogen activator Except for the above, the enzyme activity (plasmin activity) of each Bifidobacterium was calculated in the same manner as in Test Example 1 above.
（３）結果
　各ビフィドバクテリウム属細菌の蛍光強度及び酵素活性（プラスミン活性）は、表２のようになり、全てのビフィドバクテリウム属細菌が、プラスミン活性を示すことが確認された。中でもBifidobacterium longum subsp. longum ATCC BAA-999、Bifidobacterium bifidum ATCC29521、Bifidobacterium breve BCCM LMG23729、Bifidobacterium breve FERM BP-11175等の乳幼児由来のビフィドバクテリウム属細菌のプラスミン活性は、ＨＲＢの中でもとりわけ高く、ｎｏｎ‐ＨＲＢのBifidobacterium pseudolongum subsp. globosum JCM5820のプラスミン活性と比べると、いずれも６倍以上を示し、強いプラスミン活性を有することが示唆された。 (3) Results The fluorescence intensity and enzyme activity (plasmin activity) of each Bifidobacterium genus were as shown in Table 2, and it was confirmed that all Bifidobacterium bacteria exhibited plasmin activity. Among them, Bifidobacterium longum subsp. Longum ATCC BAA-999, Bifidobacterium bifidum ATCC29521, Bifidobacterium breve BCCM LMG23729, Bifidobacterium breve FERM BP-11175 and other Bifidobacterium genus bacteria have particularly high plasmin activity among HRB, and non- Compared with the plasmin activity of Bifidobacterium pseudolongum subsp. Globosum JCM5820 of HRB, all showed 6 times or more, suggesting that it has strong plasmin activity.
Figure JPOXMLDOC01-appb-T000002
Figure JPOXMLDOC01-appb-T000002
［試験例３］
　さらに、ＨＲＢであるBifidobacterium breve FERM BP-11175と、ｎｏｎ‐ＨＲＢであるBifidobacterium animalis subsp. lactis DSM10140とを用いて、それぞれプラスミン活性の高い画分を特定する試験を行った。 [Test Example 3]
Furthermore, a test for identifying fractions having high plasmin activity was performed using Bifidobacterium breve FERM BP-11175, which is an HRB, and Bifidobacterium animalis subsp. Lactis DSM10140, which is a non-HRB.
（１）培養液の調製
　試験例１の（１）と同様の手順で、前記２種のビフィドバクテリウム属細菌の培養液を調製した。 (1) Preparation of culture solution By the same procedure as in (1) of Test Example 1, culture solutions of the above-mentioned two types of Bifidobacterium were prepared.
（２）超音波破砕処理
　（１）にて調製した各培養液を４℃、５０００×ｇの条件下にて３０分間遠心処理した後、上清を捨て、分離した菌体をＰＢＳ溶液に懸濁して「菌体懸濁液」を得た。各懸濁液は、BRANSON SONIFIER 250（Branson Ultrasonics社製）を用いて超音波処理により菌体の細胞を破砕した後、１６０００×ｇ、４℃で３０分間遠心分離した。その後、上清を「水溶性画分」として回収し、沈殿物はＰＢＳ溶液に再度懸濁し「沈殿再懸濁液」として得た。 (2) Ultrasonic crushing treatment Each culture solution prepared in (1) was centrifuged at 4 ° C. and 5000 × g for 30 minutes, and then the supernatant was discarded, and the separated cells were suspended in a PBS solution. It became cloudy to obtain a “bacterial cell suspension”. Each suspension was subjected to sonication using BRANSON SONIFIER 250 (manufactured by Branson Ultrasonics), and the cells of the cells were crushed by centrifugation at 16000 × g and 4 ° C. for 30 minutes. Thereafter, the supernatant was recovered as a “water-soluble fraction”, and the precipitate was resuspended in a PBS solution to obtain a “precipitation resuspension”.
（３）プラスミン活性の測定
　（２）にて回収した水溶性画分、及び沈殿再懸濁液について、試験例２の（２）の手順にて酵素活性を測定した。 (3) Measurement of plasmin activity The enzyme activity of the water-soluble fraction collected in (2) and the precipitate resuspension was measured by the procedure of Test Example 2 (2).
（４）結果
　各サンプルの酵素活性は、表３のとおりになった。いずれのビフィドバクテリウム属細菌についても、沈殿再懸濁液の酵素活性より水溶性画分の酵素活性の方が高いことが確認された。すなわち、当該水溶性画分が顕著な非コラーゲン性糖タンパク質分解活性を有することが示唆された。また、上記２種の水溶性画分間では、ＨＲＢであるBifidobacterium breve FERM BP-11175の活性の方が、ｎｏｎ‐ＨＲＢであるBifidobacterium animalis subsp. lactis DSM10140の活性よりも高く、前者は後者の２６倍となった。 (4) Results The enzyme activity of each sample was as shown in Table 3. For any Bifidobacterium, it was confirmed that the enzymatic activity of the water-soluble fraction was higher than the enzymatic activity of the precipitate resuspension. That is, it was suggested that the water-soluble fraction has a remarkable non-collagenous glycoprotein degradation activity. Moreover, in the two water-soluble fractions, the activity of Bifidobacterium breve FERM BP-11175, which is HRB, is higher than the activity of Bifidobacterium animalis subsp. Lactis DSM10140, which is non-HRB, and the former is 26 times the latter. It became.
Figure JPOXMLDOC01-appb-T000003
Figure JPOXMLDOC01-appb-T000003
　以上のことから、本発明に係るビフィドバクテリウム属細菌及び／又はビフィドバクテリウム属細菌の培養物は、プラスミノーゲンアクチベーター活性及び／又はプラスミン活性を示すことが確認されたため、非コラーゲン糖タンパク質分解効果を有することが示唆された。そして、プラスミノーゲンアクチベーター活性及びプラスミン活性のいずれもが高かったBifidobacterium bifidum ATCC29521、Bifidobacterium breve FERM BP-11175、Bifidobacterium longum subsp. longum ATCC BAA-999及びBifidobacterium breve ATCC15700は、いずれも乳幼児由来のＨＲＢであったため、ＨＲＢの中でも乳幼児由来のＨＲＢは、顕著な非コラーゲン糖タンパク質分解効果を有することが示唆された。 From the above, it was confirmed that the Bifidobacterium bacterium and / or the Bifidobacterium culture according to the present invention exhibited plasminogen activator activity and / or plasmin activity. It was suggested to have a glycoprotein degradation effect. Bifidobacterium bifidum ATCC29521, Bifidobacterium breve FERM BP-11175, Bifidobacterium longum subsp. Longum BATCC BAA-999 and Bifidobacterium breve ATCC15700, both of which have high plasminogen activator activity and plasmin activity, are HRB derived from infants Therefore, among the HRBs, it was suggested that HRB derived from infants has a remarkable non-collagen glycoprotein degradation effect.
［製造例１］
　試験例１で用いた１７種のビフィドバクテリウム属細菌から選択される一又は複数をそれぞれ又は同一のＭＲＳ液体培地３ｍＬに添加し、３７℃で１６時間嫌気培養し、培養液を濃縮し、凍結乾燥を行い、当該一又は複数の細菌の菌末を得る。当該一又は複数の菌末と賦形剤等とを適宜混合して錠剤化する。当該錠剤を、菌の摂取量が総量で１×１０６～１×１０１２ｃｆｕ／ｋｇ体重／日になるように、３ヶ月間毎日摂取する。
　当該錠剤の摂取により、非コラーゲン性糖タンパク質分解効果が期待できる。 [Production Example 1]
One or more selected from 17 kinds of Bifidobacterium used in Test Example 1 was added to 3 mL of each or the same MRS liquid medium, anaerobically cultured at 37 ° C. for 16 hours, and the culture solution was concentrated. Freeze-drying is performed to obtain bacterial powder of the one or more bacteria. The one or more bacterial powders and excipients are mixed as appropriate to form tablets. The tablets are taken daily for 3 months so that the total amount of bacteria taken is 1 × 10 6 to 1 × 10 12 cfu / kg body weight / day.
By taking the tablet, a non-collagenous glycoprotein degradation effect can be expected.
［製造例２］
　試験例１で用いた１７種のビフィドバクテリウム属細菌から選択される一又は複数をそれぞれ又は同一のＭＲＳ液体培地３ｍＬに添加し、３７℃で１６時間嫌気培養し、培養液を濃縮し、凍結乾燥を行い、当該一又は複数の細菌の菌末を得る。当該一又は複数の菌末を発酵乳原料に添加し、発酵乳を得る。当該発酵乳を、菌の摂取量が総量で１×１０６～１×１０１２ｃｆｕ／ｋｇ体重／日になるように、少なくとも３ヶ月毎日摂取する。
　当該発酵乳の摂取により、非コラーゲン性糖タンパク質分解効果が期待できる。 [Production Example 2]
One or more selected from 17 kinds of Bifidobacterium used in Test Example 1 was added to 3 mL of each or the same MRS liquid medium, anaerobically cultured at 37 ° C. for 16 hours, and the culture solution was concentrated. Freeze-drying is performed to obtain bacterial powder of the one or more bacteria. The one or more bacterial powders are added to the fermented milk raw material to obtain fermented milk. The fermented milk is ingested daily for at least 3 months so that the total amount of bacteria is 1 × 10 6 to 1 × 10 12 cfu / kg body weight / day.
By ingesting the fermented milk, a non-collagenous glycoprotein degradation effect can be expected.
［製造例３］
　試験例１で用いた１７種のビフィドバクテリウム属細菌から選択される一又は複数の細菌を添加した調製粉乳の製造法を下記に示す。
　脱塩牛乳乳清蛋白質粉末（ミライ社製）１０ｋｇ、牛乳カゼイン粉末（フォンテラ社製）６ｋｇ、乳糖（ミライ社製）４８ｋｇ、ミネラル混合物（富田製薬社製）９２０ｇ、ビタミン混合物（田辺製薬社製）３２ｇ、ラクチュロース（森永乳業社製）５００ｇ、ラフィノース（日本甜菜製糖社製）５００ｇ、及びガラクトオリゴ糖液糖（ヤクルト薬品工業社製）９００ｇを温水３００ｋｇに溶解し、さらに９０℃で１０分間加熱溶解し、調製脂肪（太陽油脂社製）２８ｋｇを添加して均質化する。その後、殺菌、濃縮の工程を行って噴霧乾燥し、調製粉乳約９５ｋｇを調製する。これに、試験例１で用いた１７種のビフィドバクテリウム属細菌から選択される一又は複数をそれぞれ又は同一のＭＲＳ液体培地３ｍＬに添加し、３７℃で１６時間嫌気培養し、培養液を濃縮し、凍結乾燥を行った後にでん粉で倍散して得た菌末（１．８×１０11ｃｆｕ／ｇ）１００ｇを加えてビフィズス菌・オリゴ糖配合調製粉乳約９５ｋｇを調製する。得られた調製粉乳を水に溶解して、標準調乳濃度である総固形分濃度１４％（ｗ／Ｖ）の調乳液としたとき、調乳液中のビフィズス菌数は２．７×１０9ｃｆｕ／１００ｍｌとなる。上述のようにして得られた調整粉乳を摂取することにより、非コラーゲン性糖タンパク質分解効果が期待できる。 [Production Example 3]
A method for producing a formula powder to which one or a plurality of bacteria selected from the 17 types of Bifidobacterium used in Test Example 1 are added is shown below.
Desalinated milk whey protein powder (Mirai) 10kg, Milk casein powder (Fontera) 6kg, Lactose (Mirai) 48kg, Mineral mixture (Tonda Pharmaceutical) 920g, Vitamin mixture (Tanabe Pharmaceutical) 32 g, 500 g of lactulose (manufactured by Morinaga Milk Industry Co., Ltd.), 500 g of raffinose (manufactured by Nippon Sugar Sugar Co., Ltd.), and 900 g of galactooligosaccharide liquid sugar (manufactured by Yakult Yakuhin Kogyo Co., Ltd.) are dissolved in 300 kg of hot water and further heated and dissolved at 90 ° C. for 10 minutes. Then, 28 kg of prepared fat (manufactured by Taiyo Yushi Co., Ltd.) is added and homogenized. Thereafter, sterilization and concentration steps are performed and spray-dried to prepare about 95 kg of prepared milk powder. To this, one or a plurality selected from the 17 kinds of Bifidobacterium used in Test Example 1 was added to 3 mL of each or the same MRS liquid medium, and anaerobically cultured at 37 ° C. for 16 hours. About 95 kg of bifidobacteria / oligosaccharide blended milk powder is prepared by adding 100 g of bacterial powder (1.8 × 10 11 cfu / g) obtained by concentrating and freeze-drying and then triturating with starch. When the prepared powdered milk was dissolved in water to prepare a formula having a total solid content of 14% (w / V) as a standard formula, the number of bifidobacteria in the formula was 2.7 × 10 9. cfu / 100 ml. By ingesting the adjusted milk powder obtained as described above, a non-collagenous glycoprotein degradation effect can be expected.

Claims (14)

  1. 　ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とする非コラーゲン性糖タンパク質分解用組成物。 A composition for degrading non-collagenous glycoproteins comprising a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient.
  2. 　前記非コラーゲン性糖タンパク質が、フィブリン、フィブリノゲン、フィブロネクチン、ラミニン又はプラスミノーゲンである、請求項１に記載の分解用組成物。 The composition for degradation according to claim 1, wherein the non-collagenous glycoprotein is fibrin, fibrinogen, fibronectin, laminin or plasminogen.
  3. 　前記ビフィドバクテリウム属細菌が、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＡＴＣＣ１５７０７、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＡＴＣＣ　ＢＡＡ－９９９、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＡＴＣＣ１５６９７、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＢＣＣＭ　ＬＭＧ２３７２８、ビフィドバクテリウム・ブレーベＡＴＣＣ１５７００、ビフィドバクテリウム・ブレーベＦＥＲＭ　ＢＰ－１１１７５、ビフィドバクテリウム・ブレーベＢＣＣＭ　ＬＭＧ２３７２９、ビフィドバクテリウム・ビフィダムＡＴＣＣ２９５２１、ビフィドバクテリウム・アドレセンティスＡＴＣＣ１５７０３、ビフィドバクテリウム・アンギュラツムＡＴＣＣ２７５３５、ビフィドバクテリウム・デンティウムＤＳＭ２０４３６、ビフィドバクテリウム・シュードカテヌラータムＡＴＣＣ２７９１９、ビフィドバクテリウム・アニマリス・サブスピーシーズ・ラクティスＤＳＭ１０１４０、ビフィドバクテリウム・アニマリス・サブスピーシーズ・アニマリスＡＴＣＣ２５５２７、ビフィドバクテリウム・シュードロンガム・サブスピーシーズ・グロボッサムＪＣＭ５８２０、ビフィドバクテリウム・シュードロンガム・サブスピーシーズ・シュードロンガムＡＴＣＣ２５５２６、及びビフィドバクテリウム・サーモフィルムＡＴＣＣ２５５２５からなる群から選択される一又は複数である、請求項１又は２に記載の分解用組成物。 The bacteria belonging to the genus Bifidobacterium are Bifidobacterium longum subspecies longgam ATCC 15707, Bifidobacterium longum subspecies longham ATCC BAA-999, Bifidobacterium longum subspecies infan Tis ATCC 15697, Bifidobacterium longum subspecies Infantis BCCM LMG 23728, Bifidobacterium breve ATCC 15700, Bifidobacterium breve FERM BP-11175, Bifidobacterium breve BCCM LMG 23729, Bifidobacterium Um bifidum ATCC 29521, Bifidobacterium adrecentis ATCC 15703, Bifidobac L. angularatum ATCC 27535, Bifidobacterium denthium DSM 20436, Bifidobacterium pseudocatenuratam ATCC 27919, Bifidobacterium animalis sub-species Lactis DSM 10140, Bifidobacterium animalis sub-species animalis ATCC 25527 One selected from the group consisting of Bifidobacterium pseudolongum subspecies globosum JCM5820, Bifidobacterium pseudolongham subspecies pseudolongum ATCC 25526, and Bifidobacterium thermofilm ATCC 25525 Or the composition for decomposition | disassembly of Claim 1 or 2 which is plurality.
  4. 　前記ビフィドバクテリウム属細菌が、ビフィドバクテリウム・ロンガム・サブスピーシーズ・ロンガムＢＢ５３６（ＮＩＴＥ　ＢＰ－０２６２１）、ビフィドバクテリウム・ロンガム・サブスピーシーズ・インファンティスＭ－６３（ＮＩＴＥ　ＢＰ－０２６２３）、ビフィドバクテリウム・ブレーベＭ－１６Ｖ（ＮＩＴＥ　ＢＰ－０２６２２）からなる群から選択される一又は複数である、請求項１～３のいずれか一項に記載の分解用組成物。 Bifidobacterium genus bacteria are Bifidobacterium longum subspecies longum BB536 (NITE BP-02621), Bifidobacterium longum subspecies Infatis M-63 (NITE BP-02623) The decomposition composition according to any one of claims 1 to 3, which is one or more selected from the group consisting of Bifidobacterium breve M-16V (NITE BP-02622).
  5. 　ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とする非コラーゲン性糖タンパク質分解用医薬組成物。 A pharmaceutical composition for degrading non-collagenous glycoproteins comprising a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient.
  6. 　前記医薬組成物が脳梗塞、四肢動静脈血栓症、肺梗塞、脳静脈洞血栓、網膜剥離、網膜裂傷、硝子体出血、糖尿病性硝子体出血、増殖性糖尿、加齢黄斑変性、黄斑円孔、硝子体黄斑牽引、フィブリン沈着、網膜静脈閉塞症、網膜動脈閉塞症、緑内障又は網膜色素変性症の予防及び／又は治療に用いられる、請求項５に記載の医薬組成物。 The pharmaceutical composition is cerebral infarction, limb arteriovenous thrombosis, pulmonary infarction, cerebral venous sinus thrombus, retinal detachment, retinal tear, vitreous hemorrhage, diabetic vitreous hemorrhage, proliferative diabetes, age-related macular degeneration, macular hole The pharmaceutical composition according to claim 5, which is used for prevention and / or treatment of vitreal macular traction, fibrin deposition, retinal vein occlusion, retinal artery occlusion, glaucoma or retinitis pigmentosa.
  7. 　ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を有効成分とする非コラーゲン性糖タンパク質分解用飲食品組成物。 A non-collagenized glycoprotein degrading food or drink composition comprising a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium as an active ingredient.
  8. 　プラスミノーゲンを含む、請求項７に記載の飲食品組成物。 The food / beverage composition of Claim 7 containing a plasminogen.
  9. 　ビフィドバクテリウム属細菌を培養する工程、及び前記培養後の培養物中に産生されたプラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質を回収する工程を含む、プラスミノーゲンアクチベーター活性及び／又はプラスミン活性を有するタンパク質の製造方法。 Plasminogen activator activity comprising culturing a Bifidobacterium bacterium, and recovering a protein having plasminogen activator activity and / or plasmin activity produced in the culture after the culture And / or a method for producing a protein having plasmin activity.
  10. 　プラスミノーゲンを含む培地中でビフィドバクテリウム属細菌を培養する工程、及び前記培養後の培養物中に産生されたプラスミンを回収する工程を含む、プラスミンの製造方法。 A method for producing plasmin, comprising a step of culturing Bifidobacterium in a medium containing plasminogen and a step of recovering plasmin produced in the culture after the culture.
  11. 　非コラーゲン性糖タンパク質分解用組成物の製造における、ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物の使用。 Use of a Bifidobacterium genus bacterium, a culture of the bacterium, and / or a treated product of the bacterium in the manufacture of a composition for degrading non-collagenous glycoprotein.
  12. 　非コラーゲン性糖タンパク質分解に用いられるビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物。 Bifidobacterium genus used for non-collagenous glycoprotein degradation, culture of the bacterium, and / or treated product of the bacterium.
  13. 　非コラーゲン性糖タンパク質分解のためのビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物の使用。 Use of a Bifidobacterium genus, a culture of the bacterium, and / or a treated product of the bacterium for non-collagenous glycoprotein degradation.
  14. 　ビフィドバクテリウム属細菌、前記細菌の培養物、及び／又は前記細菌の菌体処理物を哺乳動物に投与する段階を含む、非コラーゲン性糖タンパク質を分解する方法。 A method for degrading non-collagenous glycoprotein, comprising a step of administering a Bifidobacterium bacterium, a culture of the bacterium, and / or a treated product of the bacterium to a mammal.
PCT/JP2018/011923 2017-03-28 2018-03-23 Composition for degrading non-collagenous glycoprotein WO2018181071A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2019509744A JP6954995B2 (en) 2017-03-28 2018-03-23 Composition for non-collagen glycoprotein degradation

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2017063417 2017-03-28
JP2017-063417 2017-03-28

Publications (1)

Publication Number Publication Date
WO2018181071A1 true WO2018181071A1 (en) 2018-10-04

Family

ID=63677653

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2018/011923 WO2018181071A1 (en) 2017-03-28 2018-03-23 Composition for degrading non-collagenous glycoprotein

Country Status (2)

Country Link
JP (1) JP6954995B2 (en)
WO (1) WO2018181071A1 (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020116511A1 (en) * 2018-12-07 2020-06-11 森永乳業株式会社 Composition for suppressing norovirus infection
JP2020164492A (en) * 2019-03-29 2020-10-08 森永乳業株式会社 Method for producing indole lactic acid compound
JP2020162479A (en) * 2019-03-29 2020-10-08 株式会社キティー Composition for improving eye trouble and use thereof
JP6978621B1 (en) * 2021-03-12 2021-12-08 森永乳業株式会社 Composition

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003238423A (en) * 2002-02-15 2003-08-27 Morinaga Milk Ind Co Ltd Hypocholesterolemic agent and hypocholesterolemic food and beverage
JP2008519018A (en) * 2004-11-05 2008-06-05 ケンブリッジ セラノスティックス リミテッド Bacterial composition for prevention or treatment of atherosclerotic disease
CN101579361A (en) * 2009-05-18 2009-11-18 薛松晓 Complex probiotics for curing hypertension and coronary heart disease and preparation method thereof
JP2011132188A (en) * 2009-12-25 2011-07-07 Kirin Holdings Co Ltd Lactobacillus having effect of enhancing activation of plasminogen and composition thereof
JP2016519063A (en) * 2013-03-14 2016-06-30 ウェイン ステイト ユニバーシティー Methods for enhancing ocular delivery of therapeutic compounds

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9730968B2 (en) * 2008-04-17 2017-08-15 Anaeropharma Science, Inc. Therapeutic agent for ischemic diseases
US20150320808A1 (en) * 2009-06-19 2015-11-12 Dupont Nutrition Biosciences Aps Bifidobacteria for treating cardiac conditions

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003238423A (en) * 2002-02-15 2003-08-27 Morinaga Milk Ind Co Ltd Hypocholesterolemic agent and hypocholesterolemic food and beverage
JP2008519018A (en) * 2004-11-05 2008-06-05 ケンブリッジ セラノスティックス リミテッド Bacterial composition for prevention or treatment of atherosclerotic disease
CN101579361A (en) * 2009-05-18 2009-11-18 薛松晓 Complex probiotics for curing hypertension and coronary heart disease and preparation method thereof
JP2011132188A (en) * 2009-12-25 2011-07-07 Kirin Holdings Co Ltd Lactobacillus having effect of enhancing activation of plasminogen and composition thereof
JP2016519063A (en) * 2013-03-14 2016-06-30 ウェイン ステイト ユニバーシティー Methods for enhancing ocular delivery of therapeutic compounds

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
AHN, M. J. ET AL.: "Characterization of a Novel Fibrinolytic Enzyme, BsfA, from Bacillus subtllls ZA 400 in Klmcbl Reveals Its Pertinence to Thrombosis Treatment.", JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, vol. 25, no. 12, 2015, pages 2090 - 2099, XP055558589 *
CANDELA, M. ET AL.: "Plasminogen-dependent proteolytic activity in Bifidobacterium lactis.", MICROBIOLOGY, vol. 154, no. 8, 2008, pages 2457 - 2462, XP055558592 *
JEFFRIES, L. ET AL.: "The detection and differentiation of fibrinolytic enzymes in bacteria.", JOURNAL OF APPLIED BACTERIOLOGY, vol. 49, no. 3, December 1980 (1980-12-01), pages 479 - 492, XP055558584 *
VASTANO, V. ET AL.: "Identification of binding sites of Lactobacillus plantarum enolase involved in the interaction with human plasminogen.", MICROBIOLOGICAL RESEARCH, vol. 168, no. 2, February 2013 (2013-02-01), pages 65 - 72, XP055558597 *
WEI, X. ET AL.: "Proteomic analysis of the interaction of Bifidobacterium longum NCC2705 with the intestine cells Caco-2 and identification of plasminogen receptors.", JOURNAL OF PROTEOMICS, vol. 108, 2014, pages 89 - 98, XP029037235 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020116511A1 (en) * 2018-12-07 2020-06-11 森永乳業株式会社 Composition for suppressing norovirus infection
EP3892331A4 (en) * 2018-12-07 2022-09-07 Morinaga Milk Industry Co., Ltd. Composition for suppressing norovirus infection
JP2020164492A (en) * 2019-03-29 2020-10-08 森永乳業株式会社 Method for producing indole lactic acid compound
JP2020162479A (en) * 2019-03-29 2020-10-08 株式会社キティー Composition for improving eye trouble and use thereof
JP6978621B1 (en) * 2021-03-12 2021-12-08 森永乳業株式会社 Composition
WO2022191320A1 (en) * 2021-03-12 2022-09-15 森永乳業株式会社 Composition
JP2022139601A (en) * 2021-03-12 2022-09-26 森永乳業株式会社 Composition

Also Published As

Publication number Publication date
JPWO2018181071A1 (en) 2020-04-16
JP6954995B2 (en) 2021-10-27

Similar Documents

Publication Publication Date Title
JP6954995B2 (en) Composition for non-collagen glycoprotein degradation
JP6856743B2 (en) Composition for Opioid Peptide Degradation
JP6088821B2 (en) Survivability improver for lactic acid bacteria and / or bifidobacteria
WO2019117212A1 (en) Composition containing bacterium belonging to genus bifidobacterium as active ingredient
EP2270133A1 (en) Method for obtaining a novel strain of bifidobacterium bifidum with activity against infection by helicobacter pylori
AU2017438436B2 (en) Muscle-building composition
JP5688376B2 (en) Oral DNA damage repair promoter and elastase activity inhibitor
AU2019243093A1 (en) Nutritional composition, food/drink composition using nutritional composition, and modified milk powder using nutritional composition
US20230066928A1 (en) Novel Bifidobacterium Genus Bacterium
Prosekov et al. Antioxidant and antimicrobial activity of bacteriocin-producing strains of lactic acid bacteria isolated from the human gastrointestinal tract
JP2019116423A (en) Composition for intestinal regulation
WO2016080371A1 (en) Lactic acid bacterium having purine body uptake ability, and use thereof
JP5209294B2 (en) Interleukin 12 production promoter
JP5603036B2 (en) Probiotic growth promoter
WO2019188943A1 (en) Composition for preventing and/or ameliorating decrease in brain blood flow
WO2020116511A1 (en) Composition for suppressing norovirus infection
WO2021059894A1 (en) Composition, food/beverage composition that contains said composition, and modified milk
WO2017047776A1 (en) Blood tryptophan concentration elevation inhibitor
JP7309436B2 (en) A composition for preventing or improving renal dysfunction, and pharmaceutical compositions and food and drink compositions using the composition for preventing or improving renal dysfunction
KR20160037879A (en) Carrier for transporting substance to macrophages

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: 18776391

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2019509744

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18776391

Country of ref document: EP

Kind code of ref document: A1