阅读说明：本技术 阿片肽分解用组合物 (Composition for decomposing opioid peptides ) 是由 樱井琢磨 桥仓那波 清水金忠 山田明男 于 2018-03-23 设计创作，主要内容包括：本发明的目的在于提供对饮食习惯没有制限、即使日常利用也没有副作用、能够安心地持续利用的阿片肽分解剂或阿片肽分解用组合物。通过以双歧杆菌属细菌、前述细菌的培养物和/或前述细菌的菌体处理物作为有效成分的阿片肽分解剂或阿片肽分解用组合物、阿片肽分解用药物组合物或阿片肽分解用饮食品组合物来解决该课题。(The purpose of the present invention is to provide an opiate peptide decomposition agent or a composition for opiate peptide decomposition that is not limited in eating habits, has no side effects even in daily use, and can be continuously used with ease. The problem is solved by an opioid peptide-decomposing agent or a composition for opioid peptide decomposition, a pharmaceutical composition for opioid peptide decomposition, or a food or beverage composition for opioid peptide decomposition, which contains a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated product of the bacterium as an active ingredient.)

1. A composition for decomposing opioid peptide, which comprises a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium and/or a treated cell product of the bacterium as an active ingredient.

2. The composition for opioid peptide degradation according to claim 1, wherein the opioid peptide is β -casomorphin.

3. The composition for decomposing opioid peptide according to claim 1 or 2, wherein the treated microbial cell product of a bacterium belonging to the genus Bifidobacterium is a water-soluble fraction of a disrupted cell product of the bacterium.

4. The opioid peptide decomposition composition according to any one of claims 1 to 3, wherein the bacterium belonging to the genus Bifidobacterium is resident in infants.

5. The opioid peptide decomposition composition according to any one of claims 1 to 4, wherein the Bifidobacterium bacteria is one or more selected from the group consisting of Bifidobacterium longum subspecies ATCC15707, Bifidobacterium longum subspecies ATCC BAA-999, Bifidobacterium longum subspecies infantile ATCC15697, Bifidobacterium longum subspecies Bifidobacterium infantis BCCM LMG23728, Bifidobacterium breve ATCC15700, Bifidobacterium breve FERM BP-11175, Bifidobacterium breve BCCM LMG23729, Bifidobacterium bifidum ATCC29521, Bifidobacterium adolescentis ATCC15703, Bifidobacterium keratinocyte ATCC27535, Bifidobacterium odonta DSM20436, Bifidobacterium pseudocatenulatum ATCC27919, Bifidobacterium animalis subspecies lactis 10140, Bifidobacterium animalis subspecies ATCC25527, Bifidobacterium pseudolongum subspecies JCM5820, Bifidobacterium pseudolongum thermophilum ATCC25526 and Bifidobacterium pseudo25525.

6. The opioid peptide decomposition composition according to any one of claims 1 to 4, wherein the Bifidobacterium bacteria is one or more selected from the group consisting of Bifidobacterium longum subspecies BB536(NITE BP-02621), Bifidobacterium longum subspecies infantis M-63(NITEBP-02623), and Bifidobacterium breve M-16V (NITE BP-02622).

7. A pharmaceutical composition for decomposing opioid peptides, which comprises a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium and/or a treated cell product of the bacterium as an active ingredient.

8. The pharmaceutical composition according to claim 7, wherein the pharmaceutical composition is for the prevention and/or treatment of autism, asperger's syndrome, Rett syndrome, childhood disintegrative disorder or sleep apnea syndrome.

9. A food or beverage composition for decomposing opioid peptides, which contains a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell of the bacterium as an active ingredient.

10. Use of a bacterium belonging to the genus Bifidobacterium, a culture of said bacterium and/or a treated product of the bacterial cell in the manufacture of a composition for opioid peptide degradation.

11. A bacterium belonging to the genus Bifidobacterium for decomposing opioid peptides, a culture of the bacterium, and/or a treated cell of the bacterium.

12. Use of a bacterium of the genus bifidobacterium, a culture of said bacterium and/or a treated product of the bacterial body of said bacterium for decomposing opioid peptides.

13. A method of decomposing an opioid peptide comprising the steps of: administering to a mammal a bacterium of the genus Bifidobacterium, a culture of said bacterium and/or a treated thallus of said bacterium.

Technical Field

The present invention relates to an opioid peptide-degrading agent or an opioid peptide-degrading composition, an opioid peptide-degrading pharmaceutical composition, and an opioid peptide-degrading food or drink composition.

Background

Among peptides are opioid peptides that bind to opioid receptors and exhibit various physiological activities. As opioid peptides, there are also known a large number of food-derived opioid peptides, for example, milk-derived opioid peptides called β -casomorphin are produced from casein as a milk protein in the digestive tract of humans. The opioid peptides are known to exert various adverse effects on organisms when not decomposed in the digestive tract. For example, it is known that opioid peptides derived from food act on cells in the intestinal tract to inhibit the absorption of cysteine, thereby reducing the antioxidant ability (non-patent document 1).

In order to avoid the above-mentioned influence of opioid peptides, for example, diets based on materials that do not produce opioid peptides in the digestive tract have been proposed (non-patent document 2). Also disclosed is an exogenous opioid peptide-digesting enzyme preparation which is intended to aid in digestion of an opioid peptide derived from a food product, which comprises at least one component selected from the group consisting of an enzyme preparation derived from Penicillium citrinum, an enzyme preparation derived from Aspergillus oryzae, and an enzyme preparation derived from Aspergillus melleus, and which exhibits a digesting activity on an opioid peptide derived from wheat gluten and an opioid peptide derived from casein (patent document 1). In addition, it is reported that: the enzyme derived from lactococcus lactis cremoris decomposes β -casomorphin, which is an opioid peptide derived from casein (non-patent document 3).

On the other hand, it is disclosed that bifidobacterium longum IATA-ES1 is used for treating food allergy because it has a hydrolysis activity of prolamin which is a gluten peptide (patent document 2). However, there have been no reports that bacteria of the genus bifidobacterium, cultures of the aforementioned bacteria, and processed products of the aforementioned bacteria have an opioid peptide-decomposing effect.

Disclosure of Invention

Problems to be solved by the invention

Diets based on materials that do not produce opioid peptides in the digestive tract are concerned with the problem that dietary habits are significantly restricted.

The purpose of the present invention is to provide an opiate peptide decomposition agent or a composition for opiate peptide decomposition that is not limited in eating habits, has no side effects even in daily use, and can be continuously used with ease.

Means for solving the problems

The present inventors have conducted intensive studies and, as a result, have found that: the present inventors have completed the present invention by finding that a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell of the bacterium has the same enzymatic activity as dipeptidyl peptidase-4 (DPP-4), and that the dipeptidyl peptidase-4 has a decomposition activity for opioid peptides.

That is, the present invention is an opioid peptide-degrading agent or a composition for opioid peptide degradation, which contains a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium as an active ingredient.

In a preferred embodiment of the opioid peptide-degrading agent or the composition for opioid peptide degradation, the opioid peptide is β -casomorphin.

In a preferred embodiment of the opioid peptide-degrading agent or the composition for opioid peptide degradation, the treated microbial cell product of a bacterium belonging to the genus Bifidobacterium is a water-soluble fraction of a disrupted cell product of the bacterium.

In a preferred embodiment of the opioid peptide-degrading agent or the composition for opioid peptide degradation, the bacterium belonging to the genus bifidobacterium is human-resident. Further, in a preferred embodiment of the opioid peptide-degrading agent or the composition for opioid peptide degradation, the bacterium belonging to the genus bifidobacterium is resident in infants.

In addition, the preferred embodiment of the opioid peptide-disrupting agent or the composition for disrupting an opioid peptide is that the bacterium belonging to the genus Bifidobacterium is selected from Bifidobacterium longum ATCC15707(Bifidobacterium longum subsp. longum ATCC15707), Bifidobacterium longum ATCC BAA-999(Bifidobacterium longum subsp. longum ATCC BAA-999) or Bifidobacterium longum BB536(NITE BP-02621) (Bifidobacterium longum subsp. longum BB536(NITE BP-02621)), Bifidobacterium longum ATCC15697(Bifidobacterium longum subsp. infantis ATCC15697), Bifidobacterium infantis CMBCG 23728(Bifidobacterium longum BCM 23728) (Bifidobacterium longum BCM 357275), Bifidobacterium breve BCM 357263 (Bifidobacterium breve BP-BCM 3575), Bifidobacterium breve ATCC 15775 (Bifidobacterium longum BCM-BCM 11133), Bifidobacterium breve BCM 15775 (Bifidobacterium longum BCM undium BCM 11133), Bifidobacterium breve BCM 15784 (Bifidobacterium longum BCM 11175), Bifidobacterium breve BCM 11133 (Bifidobacterium longum BCM 15775), Bifidobacterium longum BCM 11133 (Bifidobacterium longum BCM 15775) or Bifidobacterium breve-Bifidobacterium longum BCM 7275) (Bifidobacterium longum BCM 1578) (Bifidobacterium longum BCM 15775) (Bifidobacterium longum BCM undi) or Bifidobacterium longum BCM 1578 (Bifidobacterium longum BCM 1578) (Bifidobacterium longum BCM undi) or Bifidobacterium longum breve M-16V (NITE BP-02622)), Bifidobacterium bifidum ATCC29521(Bifidobacterium bifidum ATCC29521), Bifidobacterium adolescentis ATCC15703(Bifidobacterium adolescentis ATCC15703), Bifidobacterium horn ATCC27535(Bifidobacterium angulus ATCC27535), Bifidobacterium odonta DSM20436(Bifidobacterium bifidum DSM20436), Bifidobacterium pseudocatenulatum ATCC27919(Bifidobacterium pseudocatenulatum ATCC27919), Bifidobacterium animalis Lactobacillus lactis DSM10140(Bifidobacterium animalis DSM10140), Bifidobacterium animalis subspecies ATCC25527(Bifidobacterium animalis subsp.sp.animalis ATCC25527), Bifidobacterium longum JCM5820(Bifidobacterium anomalis Bifidobacterium longum ATCC25526), Bifidobacterium longum ATCC25525(Bifidobacterium anomalis ATCC25525), Bifidobacterium longum ATCC25525(Bifidobacterium longum ATCC25525), Bifidobacterium longum ATCC 255820 (Bifidobacterium longum ATCC 25252525525), Bifidobacterium longum ATCC 252525526, or Bifidobacterium longum ATCC 252525525 (Bifidobacterium longum).

Further, the present invention is a pharmaceutical composition for decomposing opioid peptides, which contains a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium as an active ingredient.

The aforementioned pharmaceutical composition is preferably used for the prevention and/or treatment of autism, asperger's syndrome, Rett syndrome, childhood disintegrative disorder or sleep apnea syndrome.

The present invention also provides a food or beverage composition for decomposing opioid peptides, which contains a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium as an active ingredient.

The present invention also relates to the use of a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated product of the bacterium in the production of a composition for decomposing opioid peptides.

The present invention also provides a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium, for decomposing opioid peptides.

The present invention also provides use of a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated product of the bacterium for decomposing opioid peptides.

In addition, the present invention is a method of decomposing opioid peptides comprising the steps of: administering a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell of the bacterium to a mammal.

ADVANTAGEOUS EFFECTS OF INVENTION

The present invention provides an opiate peptide decomposer or a composition for opiate peptide decomposition, a pharmaceutical composition for opiate peptide decomposition and a food and drink composition for opiate peptide decomposition, which contain as an active ingredient a bacterium belonging to the genus bifidobacterium, a culture of the bacterium and/or a treated cell product of the bacterium, wherein the bacterium belonging to the genus bifidobacterium, the culture of the bacterium and/or the treated cell product of the bacterium are used as an oral composition ingredient for a long period of time, and therefore, the administration of the composition to patients suffering from various diseases can be carried out with ease. In addition, since bacteria belonging to the genus bifidobacterium are also present in the intestinal tract of animals, it is expected that side effects are not likely to occur even if they are administered continuously for a long period of time.

Therefore, according to the present invention, it is possible to provide an opiate peptide decomposition agent or composition for opiate peptide decomposition, a pharmaceutical composition for opiate peptide decomposition, and a food and drink composition for opiate peptide decomposition, which are not limited in eating habits, have no side effect even in daily use, and can be continuously used with ease. Further, the use of the opioid peptide-degrading agent, the composition for degrading opioid peptide, and the food or beverage composition for degrading opioid peptide can suppress the direct absorption of opioid peptide mainly derived from food into the digestive tract without being degraded.

Detailed Description

Next, preferred embodiments of the present invention will be described. However, the present invention is not limited to the preferred embodiments described below, and can be freely modified within the scope of the present invention. In the present specification, percentages are expressed on a mass basis unless otherwise specified.

< opioid peptide-decomposing agent or composition for opioid peptide decomposition >

The opioid peptide-degrading agent of the present invention mainly degrades opioid peptides derived from food and contains, as an active ingredient, a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium. The opioid peptide-degrading agent of the present invention contains a bacterium belonging to the genus bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium as an active ingredient, and does not exclude other ingredients. That is, the opioid peptide-degrading agent of the present invention is equivalent to the composition for opioid peptide degradation. Accordingly, another embodiment of the present invention is a composition for opioid peptide degradation comprising a bacterium belonging to the genus bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium as an active ingredient.

In addition, the content of bacteria belonging to the genus Bifidobacterium in the composition is preferably 1 × 10⁶～1×10¹²CFU/g or 1 × 10⁶～1×10¹²CFU/mL, more preferably 1 × 10⁷～1×10¹¹CFU/g or 1 × 10⁷～1×10¹¹CFU/mL, more preferably 1 × 10/mL⁸～1×10¹⁰CFU/g or 1 × 10⁸～1×10¹⁰CFU/mL. When the bacteria are dead, the CFU can be replaced with individual cells (cells).

The bacterium belonging to the genus Bifidobacterium, the culture of the bacterium, and the treated product of the bacterial cell of the bacterium of the present invention have the same enzymatic activity as Dipeptidyl Peptidase-4 (DPP-4) which is one of Dipeptidyl aminopeptidases.

DPP-4 is an exopeptidase having an action of cleaving a dipeptide from the amino-side terminus of a protein or peptide, and is an enzyme having an activity of degrading casomorphins as opioid peptides (G.Puschel et al, Eur.J.biochem.,126(2), pp.359-365, 1982).

In the present specification, "DPP-4 activity" means an enzyme activity similar to or the same as DPP-4, specifically, an activity of degrading a DPP-4-specific substrate. Examples of the DPP-4-specific substrate include proteins and peptides each having a proline residue or an alanine residue at position 2 from the amino terminus.

Another embodiment of the present invention is a method for producing a composition for opioid peptide degradation, which comprises adding the bacterium belonging to the genus bifidobacterium of the present invention to a raw material of the composition. The method for producing a food or beverage composition comprises a step of adding the bacterium belonging to the genus Bifidobacterium of the present invention to a raw material of a food or beverage composition. The production method also includes a method for producing a pharmaceutical composition, which comprises a step of adding a bacterium belonging to the genus Bifidobacterium to a raw material (e.g., a base material or the like) of the pharmaceutical composition.

The bacterium belonging to the genus bifidobacterium used in the method for producing the composition for opioid peptide degradation may be a living bacterium or a dead bacterium, or may contain both of the living bacterium and the dead bacterium. Further, the microorganism may be a culture of bacteria belonging to the genus Bifidobacterium or a treated product of the microorganism. In the method for producing the composition for opioid peptide degradation, the bacterium belonging to the genus bifidobacterium may be added to the raw material of the composition after culturing, concentrating or freeze-drying the bacterium, or the bacterium belonging to the genus bifidobacterium may be cultured after adding the bacterium belonging to the genus bifidobacterium to the raw material of the composition.

(1) Bacterium belonging to the genus Bifidobacterium

The bacterium belonging to the genus bifidobacterium that can be used in the present invention may be any known bacterium belonging to the genus bifidobacterium as long as the effect of the present invention is not impaired. For example, one can cite: bifidobacterium longum subsp. species (Bifidobacterium longum subsp. sp. longum), Bifidobacterium longum subsp. infantis (Bifidobacterium longum subsp. infantis), Bifidobacterium breve (Bifidobacterium breve), Bifidobacterium bifidum (Bifidobacterium bifidum), Bifidobacterium adolescentis (Bifidobacterium adolescentis), Bifidobacterium angulus (Bifidobacterium angulus), Bifidobacterium pseudocatenulatum (Bifidobacterium pseudocatenulatum), Bifidobacterium animalis (Bifidobacterium animum lactis), Bifidobacterium animalis subsp. animalis (Bifidobacterium animalis), Bifidobacterium pseudocatenulatum (Bifidobacterium longum subsp. lactis), Bifidobacterium pseudocatenulatum (Bifidobacterium longum subsp. sp. thermophilum), Bifidobacterium pseudocatenulatum (Bifidobacterium longum subsp. sp. Bifidobacterium longum), Bifidobacterium pseudobulb (Bifidobacterium longum) and the like. Incidentally, Bifidobacterium longum subspecies longum may be abbreviated as Bifidobacterium longum. In addition, bifidobacterium longum subspecies infantis is sometimes also abbreviated as bifidobacterium infantis.

Specifically, it is possible to exemplify: bifidobacterium longum strain ATCC15707(Bifidobacterium longum. sp. longum ATCC15707), Bifidobacterium longum strain ATCC BAA-999(Bifidobacterium longum. sp. ATCC BAA-999) or Bifidobacterium longum strain BB536(NITE BP-02621) (Bifidobacterium longum. sp. BB536(NITE BP-02621)), Bifidobacterium longum strain ATCC15697(Bifidobacterium longum. sp. infantis ATCC15697), Bifidobacterium infantis BCCM 728(Bifidobacterium longum. infantis BCG 23728) or Bifidobacterium longum strain M-63(NITE BP-02623) (Bifidobacterium longum. sp. ATCC 15775), Bifidobacterium longum strain ATCC 7275 (Bifidobacterium longum. sp. illusm-02684) (Bifidobacterium longum. sp. illusm-02675), Bifidobacterium longum strain ATCC 15775 (Bifidobacterium longum BCM 11184), Bifidobacterium longum strain ATCC 15775 (Bifidobacterium longum M-BCM 11175) or Bifidobacterium breve strain ATCC 15716 (Bifidobacterium longum sp. 02675) (Bifidobacterium sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.22) or Bifidobacterium sp. sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.sp.15775 (Bifidobacterium sp.sp.sp.sp.sp.sp.sp.sp.sp.sp, Bifidobacterium bifidum ATCC29521(Bifidobacterium bifidum ATCC29521), Bifidobacterium adolescentis ATCC15703(Bifidobacterium adolescentis ATCC15703), Bifidobacterium horn ATCC27535(Bifidobacterium angulus ATCC27535), Bifidobacterium odonum DSM20436(Bifidobacterium denum DSM20436), Bifidobacterium pseudocatenulatum ATCC27919(Bifidobacterium pseudostellatum ATCC27919), Bifidobacterium animalis lactococcus lactis DSM10140(Bifidobacterium animalis subsp.lactis DSM10140), Bifidobacterium animalis subspecies ATCC25527(Bifidobacterium animalis subsp.sp.ATCC 25526), Bifidobacterium pseudolongum globular bacillus 5820(Bifidobacterium pseudobifidobacterium longum ATCC25526), Bifidobacterium pseudolongum Bifidobacterium longum ATCC 2525526, Bifidobacterium pseudolongum ATCC25526, etc. One species of the bacterium belonging to the genus Bifidobacterium may be used alone, or two or more species of the bacterium may be used.

Bacteria with ATCC numbers assigned thereto can be obtained from the research American type culture Collection (address: 12301Parklawn Drive, Rockville, Maryland 20852, United States of America).

The bacteria to which BCCM numbers are assigned can be obtained from Belgian Coordinated Collections of Microorganisms (address: Belgium, B-1000 Brussel Shiatsu Street (STREET CUP) 8) which is a storage organization in Belgium.

The bacteria to which the FERM BP-11175 deposit number was assigned were subjected to International deposit under the Budapest treaty at 25.8.2009 at the International patent organism depositary, national institute of advanced Industrial science and technology, International patent organism depositary, postal code 292-.

The bacteria to which the DSM numbers are assigned can be obtained from the German Collection of microorganisms DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH) (address: Inhoffenttra beta e7B,38124Braunschweig, Germany).

The bacterium to which the JCM number is assigned can be obtained from the Japanese Collection management center JCM (Japan Collection of microorganisms) (national institute of research and development, national institute of research and chemistry, institute of Bioresource, center, development of microbial materials, zip code 305-0074, and Hitachi 3-1-1, Kyowa prefecture, Japan).

Note that Bifidobacterium longum subspecies ATCC BAA-999 is the same bacterium as Bifidobacterium longum subspecies BB536(NITE BP-02621), and any bacterium can be used in a preferred embodiment. Bifidobacterium longum subspecies BB536(NITE BP-02621) was deposited under the number NITE BP-02621 on 26.1.2018, and international deposit under the Budapest treaty was made at the national institute of technology and evaluation, patent microorganism depositary (postal code: 292 + 0818 Kye county, Tokyo Gentianzu 2-5-8122).

Bifidobacterium longum subspecies infantis BCCM LMG23728 and Bifidobacterium longum subspecies infantis M-63(NITE BP-02623) are the same bacteria, and any bacteria can be used in a preferable embodiment. Bifidobacterium longum subspecies of infantis M-63(NITE BP-02623) was internationally deposited under the Budapest treaty at 26.1.2018 with the accession number NITE BP-02623 at the national institute of technical assessment, the patent microorganism Collection (postal code: 292 + 0818 Kye county, woodgun, Gentianzu 2-5-8122).

Bifidobacterium breve BCCM LMG23729 is the same bacterium as Bifidobacterium breve M-16V (NITE BP-02622), and any bacterium can be used in a preferred embodiment. Bifidobacterium breve M-16V (NITE BP-02622) was deposited at 26.1.2018 under the NITE BP-02622 deposit number at the national institute of technology and evaluation, International deposit under the Budapest treaty at the patent microorganism depositary (postal code: 292 + 0818 Kye, woodbethezu, 2-5-8122 Country).

The bacterium belonging to the genus Bifidobacterium 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 species as the deposited bacterium, and is capable of decomposing opioid peptide in a subject when the bacterium, a culture of the bacterium, and/or a treated cell product of the bacterium are ingested or administered into the subject, and the nucleotide sequence of the 16S rRNA gene of the bacterium has 98% or more, preferably 99% or more, and more preferably 100% homology with the nucleotide sequence of the 16S rRNA gene of the deposited bacterium, and is preferably a bacterium having the same bacteriological properties as the deposited bacterium except for the homology. The bacterium belonging to the genus bifidobacterium of the present invention may be a bacterium obtained by breeding the deposited bacterium or a bacterium substantially equivalent thereto by mutation treatment, gene recombination, selection of a natural mutant strain, or the like, as long as the effect of the present invention is not impaired.

Among them, bacteria belonging to the genus Bifidobacterium, which are resident in the human body, are preferably used in the present invention. Bacteria belonging to the genus Bifidobacterium live in insects and small animals in addition to humans, and the habitat (host) is limited by the species, and bacteria belonging to the genus Bifidobacterium, which mainly live in humans, are called Human Resident Bifidobacterium (HRB) and are otherwise called non-HRB. As the HRB, there can be exemplified: bifidobacterium longum (Bifidobacterium longum), Bifidobacterium longum subsp. infantis (Bifidobacterium longum subsp. infantis), Bifidobacterium breve (Bifidobacterium breve), Bifidobacterium bifidum (Bifidobacterium bifidum), Bifidobacterium adolescentis (Bifidobacterium adolescentis), Bifidobacterium horn (Bifidobacterium angium), Bifidobacterium odonum (Bifidobacterium denum), Bifidobacterium pseudocatenulatum (Bifidobacterium udocatenonum), and the like.

Further, HRB of the present invention is more preferably bacteria belonging to the genus bifidobacterium, which are resident in infants. Examples of the infant-resident HRB include: bifidobacterium longum (Bifidobacterium longum), Bifidobacterium longum subsp.

The bacterium belonging to the genus bifidobacterium used in the present invention may be a mutant strain of the bacterium belonging to the genus bifidobacterium as long as it has an opioid peptide-degrading effect equivalent to or higher than that of the bacterium belonging to the genus bifidobacterium. Whether or not a certain mutant has an opioid peptide-degrading effect equal to or greater than that of the bacterium belonging to the genus bifidobacterium can be confirmed, for example, by: measurement of DPP-4 Activity of Bifidobacterium bacteria described later; the consumption rate (decomposition rate) of β -casomorphin as opioid peptide was measured.

Such a mutant strain can be constructed by artificially introducing a mutation into the bacterium belonging to the genus Bifidobacterium. Further, the strain can be constructed by introducing a mutation into the bacterium by treatment with a mutagen such as UV, or by introducing a mutation into the strain by various gene manipulation methods.

In the present invention, DPP-4 activity can be confirmed as follows: a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium and/or a treated cell product of the bacterium are mixed with a DPP-4-specific fluorogenic substrate, and the fluorescence intensity of the cultured culture is measured. That is, the enzyme activity (DPP-4 activity) of a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium can be determined by calculating the fluorescent substance concentration in the culture using a calibration curve showing the relationship between the fluorescent substance concentration derived from the fluorogenic substrate and the fluorescent intensity based on the fluorescent intensity of the culture, and further using 1nmol of the fluorescent substance produced in 1 minute as 1 unit of enzyme (U).

The enzyme activity (DPP-4 activity) can be determined, for example, as follows: bacterial cells are separated from a culture of a bacterium belonging to the genus Bifidobacterium and suspended in a PBS solution to prepare a suspension, the suspension is diluted to have a turbidity (OD600) of 0.1, and then H-Gly-Pro-AMC-HBr (manufactured by BACHEM) as a DPP-4 specific fluorogenic substrate is added thereto and the suspension is anaerobically incubated at 37 ℃ for 60 minutes, and after the end of the incubation, the enzyme activity (DPP-4 activity) is determined based on the fluorescence intensity of the culture measured at an excitation wavelength of 380nm and a measurement wavelength of 460nm by a fluorometer such as a microplate reader SH-9000 (manufactured by Corona Electric Co., Ltd.).

In the present invention, the opioid peptide-decomposing effect of the present invention can be suitably exhibited by the DPP-4 activity as long as it is preferably 0.5mU or more, more preferably 0.7mU or more, and still more preferably 1.0mU or more.

In the present invention, the consumption rate (decomposition rate) of β -casomorphin can be calculated as follows: bacteria of the genus bifidobacterium and β -casomorphin were mixed and cultured, and the concentration of β -casomorphin in the culture was used and calculated by the following formula.

[ consumption rate (%) of β -casomorphin ] - [ 100- ([ concentration of β -casomorphin in the culture after cultivation ]/[ concentration of β -casomorphin in the culture before cultivation ]/[ 100 [ ]

Specifically, the confirmation can be performed as described in test example 3 described later.

In the present invention, the opioid peptide degradation effect of the present invention can be suitably exhibited as long as the consumption rate (degradation rate) of β -casomorphin is preferably 40% or more, more preferably 50% or more, and even more preferably 60% or more.

The bacterium belonging to the genus Bifidobacterium and the culture of the aforementioned bacterium used in the present invention can be easily obtained by culturing the bacterium belonging to the genus Bifidobacterium by a conventional method. The culture method is not particularly limited as long as it can proliferate the bacterium belonging to the genus bifidobacterium, and the culture can be carried out under suitable conditions depending on the nature of the bacterium. For example, the culture temperature may be 25 to 50 ℃ and preferably 35 to 42 ℃. The culture is preferably carried out under anaerobic conditions, and for example, the culture can be carried out while introducing an anaerobic gas such as carbon dioxide. In addition, the culture may be performed under microaerobic conditions, such as liquid static culture.

The medium for culturing bacteria of the genus bifidobacterium used in the present invention is not particularly limited, and a medium generally used in culturing bacteria of the genus bifidobacterium can be used. That is, as the carbon source, for example, sugars such as glucose, galactose, fructose, arabinose, mannose, sucrose, starch hydrolysate, molasses and the like can be used according to assimilation. As nitrogen sources, for example: ammonium salts such as ammonia, ammonium sulfate, ammonium chloride and ammonium nitrate, and nitrates. As the inorganic salts, for example, there can be used: sodium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, ferrous sulfate, and the like. In addition, it is possible to use: organic components such as peptone, soybean powder, defatted soybean meal, meat extract, and yeast extract.

The bacterium belonging to the genus Bifidobacterium used in the present invention may be used in the form of the bacterium itself, a culture of the bacterium, or a treated cell of the bacterium, as described above. That is, the bacterium belonging to the genus Bifidobacterium may be used as it is, a culture obtained by culturing the bacterium belonging to the genus Bifidobacterium may be used as it is, or a cell recovered from the culture may be used after dilution or concentration. The bacterium belonging to the genus bifidobacterium used in the present invention may be a living bacterium or a dead bacterium, or may be a substance containing both of a living bacterium and a dead bacterium.

Examples of the treated product of the bacterial cells of the above-mentioned bacteria include: immobilized cells obtained by immobilizing cells with acrylamide, carrageenan, or the like; and disrupted cell products obtained by disrupting a part or all of cell walls and cell membranes of the cells by a conventional method such as ultrasonic treatment or homogenizer treatment.

The disrupted cell product may be the whole or a part of the disrupted cell product, or may be a supernatant obtained by centrifugation after the disruption, a fraction obtained by partially purifying the supernatant by ammonium sulfate treatment or the like, or a product obtained by concentrating the supernatant.

Specific examples of the bacterium belonging to the genus Bifidobacterium used in the present invention include a suspension of the bacterium, a water-soluble fraction of a disrupted cell of the bacterium, a sediment resuspension, and the like, and among them, a water-soluble fraction of a disrupted cell of the bacterium is preferable because of its high opioid peptide-degrading activity.

The "cell suspension" in the present specification is a solution obtained by suspending cells obtained by subjecting a culture solution of bacteria belonging to the genus bifidobacterium used in the present invention to centrifugation or the like, and is preferably a solution obtained by the treatment described in (2) of test example 2 of the example described later.

The "water-soluble fraction" in the present specification is a supernatant obtained by subjecting the cell disruption product to centrifugation or the like, and is preferably a supernatant finally obtained by the treatment described in (2) of test example 2 of the examples described later. The water-soluble fraction is more preferably a water-soluble fraction obtained by suitable purification or concentration using a conventional method.

Further, the "re-suspension of the pellet" in the present specification is a solution obtained by suspending the pellet obtained by subjecting the cell debris to centrifugal separation or the like, and is preferably a solution obtained by suspending the pellet finally obtained by the treatment described in (2) of test example 2 of the example described later.

(2) Opioid peptides

Opioid peptides are peptides that bind to opioid receptors and exhibit various physiological activities. Although endogenous peptides and exogenous peptides synthesized in vivo are present in the opioid peptides, the opioid peptides of the present invention are preferably exogenous opioid peptides, and more preferably opioid peptides derived from food. As opioid peptides derived from foods, β -casomorphin, human gliadin (Gliadorphin), and the like can be exemplified.

< pharmaceutical composition >

The opioid peptide-degrading agent or the composition for opioid peptide degradation of the present invention can be used as a pharmaceutical composition. It is known that direct absorption of opioid peptides into the body without degradation has an effect on symptoms such as autism, asperger's syndrome, Rett syndrome, childhood disintegrative disorder, and the like (patent document 1). For example, it is reported that: the higher the urinary concentration of beta-casomorphin as opioid peptide in autistic children, the higher the value of the children's Autism Rating Scale (Childhood Autism Rating Scale: CARS) (O.Sokolov et al, Peptides,56, pp.68-71,2014). In addition, it is reported that: the concentration of beta-casomorphin in the blood of infants with the occurrence of sudden life threatening events in infants such as Apneas (ALTE) is high compared to healthy children and the activity of DPP-4, an enzyme that breaks down beta-casomorphin, is low compared to healthy children (J.Wasiewska et al, Nuropipetides, 45, pp.189-195,2011).

Therefore, the pharmaceutical composition of the present invention can be used for preventing and/or treating diseases related to opioid peptides. As the diseases related to opioid peptides, there can be exemplified: autism, asperger's syndrome, Rett syndrome, childhood disorganized or sleep apnea syndrome, and the like.

In addition, since the pharmaceutical composition of the present invention contains, as an active ingredient, a bacterium belonging to the genus bifidobacterium, a culture of the bacterium, and/or a processed product of the bacterium, which can be used for a long period of time as an oral composition ingredient, it can be administered to patients suffering from various diseases with ease. In addition, bacteria belonging to the genus bifidobacterium are also present in the intestinal tract of animals, and therefore, it is expected that side effects are not likely to occur even when they are continuously administered for a long period of time. Further, the opioid peptide-degrading agent or the composition for opioid peptide degradation of the present invention contains a bacterium belonging to the genus bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium as an active ingredient, and therefore can be safely administered to infants and children. Therefore, the opioid peptide-degrading agent or the composition for opioid peptide degradation of the present invention is suitable for the prevention and/or treatment of diseases in infants and children.

When the opioid peptide-degrading agent or the composition for opioid peptide degradation of the present invention is used as a pharmaceutical composition, the pharmaceutical composition may be administered orally or parenterally, and the pharmaceutical composition may be appropriately prepared into a desired dosage form according to the administration method. For example, in the case of oral administration, the preparation may be formulated into solid preparations such as powder, granule, tablet, capsule and the like; liquid preparations such as solutions, syrups, suspensions, and emulsions. In addition, in the case of parenteral administration, the preparation may be formulated into suppositories, ointments and the like.

In addition, in the case of formulation, in addition to the opioid peptide-degrading agent or the composition for opioid peptide degradation of the present invention, ingredients such as excipients, pH adjusters, coloring agents, and flavoring agents generally used in formulation may be used. The opioid peptide-degrading agent or the composition for opioid peptide degradation of the present invention and components known or discovered in the future to have a prophylactic and/or therapeutic effect on opioid peptide-related diseases may be used in combination as long as the effect of the present invention is not impaired.

The formulation may be carried out by a known method according to the dosage form. When formulated, the formulation may be formulated by appropriately mixing a formulation carrier.

The intake amount or the administration amount of the pharmaceutical composition of the present invention may be suitably selected depending on the dosage form, and for example, the intake amount or the administration amount of the bacterium belonging to the genus Bifidobacterium per 1 day is preferably 1 × 10 per 1kg body weight⁶～1×10¹²CFU/kg/day, more preferably 1 × 10⁷～1×10¹¹CFU/kg/day, more preferably 1 × 10⁸～1×10¹⁰CFU/kg/day. In the above-mentioned units, CFU is a short hand for colony forming units (colony forming units), and is a colony forming unit. When the bacteria are dead, the CFU can be replaced with individual cells (cells).

In addition, the intake amount or the administration amount when using a culture of a bacterium belonging to the genus bifidobacterium or a treated cell of the bacterium is preferably the intake amount or the administration amount in terms of the intake amount or the administration amount of the bacterium belonging to the genus bifidobacterium.

The content of bacteria belonging to the genus Bifidobacterium in the pharmaceutical composition of the present invention may be appropriately selected based on the above-mentioned intake amount or administration amount, and may be, for example, 1 × 10⁶～1×10¹²CFU/g or 1 × 10⁶～1×10¹²CFU/mL, preferably 1 × 10⁷～1×10¹¹CFU/g or 1 × 10⁷～1×10¹¹CFU/mL, more preferably 1 × 10⁸～1×10¹⁰CFU/g or 1 × 10⁸～1×10¹⁰CFU/mL. When the bacteria are dead, the CFU can be replaced with individual cells (cells).

The content of the culture of bacteria belonging to the genus bifidobacterium or the treated product of the bacteria of the genus bifidobacterium is preferably the above content in terms of the content of the bacteria belonging to the genus bifidobacterium.

As the formulation vehicle, various organic or inorganic vehicles can be used depending on the formulation. Examples of the carrier in the case of a solid preparation include an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring agent, and the like.

Examples of the excipient include: sugar derivatives such as lactose, white sugar, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α -starch, dextrin, and carboxymethyl starch; cellulose derivatives such as crystalline cellulose, hydroxypropyl methylcellulose, carboxymethyl cellulose, and calcium carboxymethyl cellulose; acacia gum; (ii) a glucan; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, magnesium aluminate metasilicate, etc.; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; sulfate derivatives such as calcium sulfate, and the like.

Examples of the binder include, in addition to the above-mentioned excipients: gelatin; polyvinylpyrrolidone; macrogol, and the like.

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 crospolyvinylpyrrolidone.

Examples of the lubricant include: talc; stearic acid; metal stearates such as calcium stearate and magnesium stearate; silica gel; colloidal magnesium aluminum silicate (VEEGUM), spermaceti wax, etc.; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid; sodium carboxylates 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 silicic anhydride and silicic acid hydrate; starch derivatives, and the like.

Examples of the stabilizer include: parabens such as methyl paraben and propyl paraben; alcohols such as chlorobutanol, benzyl alcohol, and phenethyl alcohol; benzalkonium chloride; acetic anhydride; sorbic acid, and the like.

Examples of the taste corrigent include a sweetener, an acidulant, and a perfume.

The carrier used in the liquid preparation for oral administration includes solvents such as water, flavoring agents, and the like.

< food and drink composition >

Further, the opioid peptide-degrading agent or the composition for opioid peptide degradation of the present invention can be used as a food or beverage composition. The food or beverage composition of the present invention can be produced by adding a bacterium belonging to the genus bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium to a known food or beverage, or can be produced as a novel food or beverage composition by mixing a bacterium belonging to the genus bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium with a raw material for a food or beverage. The food or beverage composition of the present invention can be produced by a production method comprising the steps of: bacteria belonging to the genus Bifidobacterium are added to food and drink materials and then cultured.

By orally ingesting a food or beverage composition containing bacteria of the genus bifidobacterium, the bacteria of the genus bifidobacterium, which proliferate in vivo (intestinal tract), can decompose opioid peptides, and thus the opioid peptides can be prevented from being directly absorbed into the body by the digestive tract without being decomposed.

The food or beverage composition of the present invention may be in the form of liquid, paste, solid, powder, etc., and may be in the form of tablet candy, liquid food, feed (including for pet), etc., for example: wheat flour products, instant foods, processed agricultural products, processed marine products, processed livestock products, milk/dairy products, fats and oils, base seasonings, compound seasonings/foods, frozen foods, snacks, beverages, and other commercial products. In addition, a component having a beneficial bacterial effect or a component assisting the beneficial bacterial effect, which is known or discovered in the future, may be used in the food or beverage composition of the present invention as long as the effect of the present invention is not impaired. For example, the food or beverage composition of the present invention may contain various proteins such as whey protein, casein protein, soybean protein, pea protein (p protein), or a mixture or a decomposed product thereof; amino acids such as leucine, valine, isoleucine, or glutamine; vitamins such as vitamin B6 and vitamin C; creatine; citric acid; fish oil; or oligosaccharide such as isomaltooligosaccharide, galactooligosaccharide, xylooligosaccharide, soybean oligosaccharide, fructooligosaccharide, and lactulose.

The food or drink composition defined in the present invention can be provided and sold in the form of a food or drink that shows uses (including health-care uses) such as prevention of an opioid peptide-related disease, reduction of the risk of a disease, alleviation of symptoms of a disease, and/or treatment of a disease.

The "marking" behavior includes all behaviors for making a user aware of the above-described use, and is equivalent to the "marking" behavior of the present invention regardless of the purpose of marking, the content of marking, the object/medium of marking, and the like as long as the expression of the above-described use can be conceived and analogized.

Further, "indication" is preferably performed in a manner such that a person in need is directly aware of the above-described use. Specifically, there are given the behaviors of transferring, delivering, displaying for transfer or delivery, and importing goods in which the use is described in commodities of foods and drinks or packages of commodities; an act of describing the above-mentioned usage in an advertisement, a price list, or a transaction document relating to a commodity and displaying or issuing the same, or describing the above-mentioned usage in information containing the same and providing the same by an electromagnetic method (the internet or the like); and the like.

On the other hand, the content of the indication is preferably an indication that permission such as administration is obtained (for example, an indication that permission is obtained based on various systems specified by administration and that permission is obtained). It is also preferable that such a label is attached to a package, a container, a catalog, a brochure, a promotional material on a sales site such as POP, or other documents.

Examples of the "label" include labels as health foods, functional foods, enteral nutritional foods, special-purpose foods, health functional foods, specific health foods, nutritional functional foods, functional label foods, and quasi drugs. Among these, mention may be made in particular of: examples of the indications permitted by the consumption hall include indications permitted by a system relating to a specific health food, a nutritional functional food, or a functional indication food, and indications permitted by a system similar to these. Specific examples thereof include: a specific health food label, a specific health food label as an additional condition, a label mainly affecting the structure and function of the body, a label for reducing the risk of diseases, a label based on the functionality of scientific grounds, and the like, and more specifically, typical examples thereof are: a label (in particular, a label for health care use) of a specific health food determined by a national condominium ( th condominium within thirty-one twenty years) as a license for a special use label prescribed in health promotion act, and the like, and a label similar thereto.

The intake amount of the food or beverage composition of the present invention can be suitably selected, and for example, the intake amount of bacteria belonging to the genus bifidobacterium per 1 day per 1kg body weight is preferably 1 × 10⁶～1×10¹²CFU/kg/day, more preferably 1 × 10⁷～1×10¹¹CFU/kg/day, more preferably 1 × 10⁸～1×10¹⁰CFU/kg/day. When the bacteria are dead, the CFU can be replaced with individual cells (cells).

The intake amount when using a culture of a bacterium belonging to the genus bifidobacterium or a treated cell of the bacterium is preferably the above intake amount in terms of the intake amount of the bacterium belonging to the genus bifidobacterium.

The content of the bacteria belonging to the genus bifidobacterium in the food or beverage composition of the present invention may be appropriately selected based on the intake amount, and may be, for example, 1 × 10⁶～1×10¹²CFU/g or 1 × 10⁶～1×10¹²CFU/mL, preferably 1 × 10⁷～1×10¹¹CFU/g or 1 × 10⁷～1×10¹¹CFU/mL, more preferably 1 × 10⁸～1×10¹⁰CFU/g or 1 × 10⁸～1×10¹⁰CFU/mL. When the bacteria are dead, the CFU can be replaced with individual cells (cells).

The content of the culture of bacteria belonging to the genus bifidobacterium or the treated product of the bacteria of the genus bifidobacterium is preferably the above content in terms of the content of the bacteria belonging to the genus bifidobacterium.

The food or beverage composition for opioid peptide degradation of the present invention can be used as a food or beverage composition for humans or animals. The food or beverage composition for opioid peptide degradation of the present invention is effective for the prevention and/or treatment of diseases related to opioid peptides, such as autism, asperger's syndrome, Rett syndrome, childhood disintegrative disorder, and sleep apnea syndrome. The food or beverage composition for opioid peptide degradation of the present invention contains a bacterium belonging to the genus bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium as an active ingredient, and therefore can be safely administered to infants and children. Therefore, the food or beverage composition for opioid peptide degradation of the present invention is also suitable for the prevention and/or treatment of diseases in infants and children.

Further, the present invention may adopt the following configuration. The mammals to be mentioned below include humans, cows, sheep, goats, pigs, dogs, cats, horses, and the like.

[1] Use of a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium and/or a treated product of the bacterium in the production of a composition for opioid peptide degradation or a drug for opioid peptide degradation.

[2] A bacterium belonging to the genus Bifidobacterium for decomposing opioid peptides, a culture of the bacterium, and/or a treated cell product of the bacterium.

[3] A bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell of the bacterium, for alleviating, preventing or treating a disease which can be alleviated, prevented or treated by the decomposition of an opioid peptide.

[4] Use of a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium and/or a treated product of the bacterium for decomposing opioid peptides.

[5] A method of decomposing an opioid peptide comprising the steps of: administering a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell of the bacterium to a mammal.

[6] A method of decomposing an opioid peptide comprising the steps of: an opioid peptide-degrading agent or an opioid peptide-degrading composition containing a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium as an active ingredient is administered to a mammal.

[7] A method of palliating, preventing or treating a disease that can be palliatively, prophylactically or therapeutically treated by the breakdown of opioid peptides comprising the steps of: administering a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell of the bacterium to a mammal.

[8] A method of palliating, preventing or treating a disease that can be palliatively, prophylactically or therapeutically treated by the breakdown of opioid peptides comprising the steps of: an opioid peptide-degrading agent or an opioid peptide-degrading composition containing a bacterium belonging to the genus Bifidobacterium, a culture of the bacterium, and/or a treated cell product of the bacterium as an active ingredient is administered to a mammal.