Ghrelin secretion-enhancing composition

Abstract

To provide a novel ghrelin secretion-enhancing composition.SOLUTION: The present invention provides a ghrelin secretion-enhancing composition containing a bacterium classified under Lactobacillus delbrueckii, and a composition used for one or more purposes selected from the group consisting of enhancement of growth hormone secretion, inhibition of brown adipose tissue function, enhancement of gastrointestinal motility, stimulation of gastric acid secretion, stimulation of pancreatic exocrine secretion, vasodilation, augmentation of myocardial contractility, and protection of cardiomyocytes.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition for promoting ghrelin secretion. [Background technology]

[0002] Ghrelin is a peptide hormone secreted primarily by endocrine cells in the stomach and has been reported to play a variety of roles, including regulating energy metabolism, promoting gastric acid secretion and gastrointestinal motility, dilating blood vessels, protecting the cardiovascular system, and having an antidiabetic effect.

[0003] Ghrelin has the effect of promoting growth hormone secretion. Growth hormone and ghrelin are peptide hormones, and if taken orally as is, they are broken down by digestive enzymes and cannot exert their effects. In addition, there are various issues with the injection of growth hormone, such as doping issues, safety, and side effects. Therefore, a method that is highly safe, can be easily taken, and can promote the endogenous secretion of growth hormone is desired.

[0004] Patent Documents 1 and 2 describe hormone secretion promoters containing skim milk or skim milk powder as active ingredients, and describe that the hormones referred to in these are ghrelin or growth hormone. Patent Document 3 describes a ghrelin secretion promoter containing whey protein hydrolysate and fermented milk protein as proteins; medium-chain fatty acids, milk phospholipids, and fish oil as lipids; and Palatinose (registered trademark) as carbohydrates. Patent Document 4 describes an anorexia improver containing Lactobacillus gasseri OLL2716 (FERM BP-6999) (reclassified as Lactobacillus paragasseri in 2020) as an active ingredient, and describes that this improvement in anorexia is due to the secretion-promoting effect of ghrelin. Patent Document 5 further describes peptides having a specific sequence of 4 to 12 amino acids derived from a subtilisin digest of purified rice endosperm protein, and describes that these peptides are effective in treating anorexia caused by a decrease in ghrelin secretion.

[0005] On the other hand, Lactobacillus delbrueckii is a kind of lactic acid bacteria. Patent Document 6 describes a lipid metabolism improver containing a lactic acid bacteria cell destruction product as an active ingredient, in which the average major axis of the cell destruction product is 0-70% of that before destruction, and lists Lactobacillus delbrueckii subsp. bulgaricus as an example of lactic acid bacteria. Patent Document 7 describes fermented milk for improving fatigue prepared using Lactobacillus lactic acid bacteria, and lists Lactobacillus delbrueckii subsp. bulgaricus OLL1073R-1 as the Lactobacillus lactic acid bacteria. Patent Document 8 describes fermented milk for improving autonomic nervous function, which contains at least one type of Lactobacillus lactic acid bacterium selected from the group consisting of Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus gasseri, Lactobacillus rhamnosus, and Lactobacillus acidophilus, and at least one of its metabolites, and describes Lactobacillus delbrueckii subsp. bulgaricus OLL1073R-1 (FERM BP-10741) as the Lactobacillus lactic acid bacterium. Patent Document 9 discloses a probiotic compound VSL that contains eight kinds of useful bacteria including Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus acidophilus, Streptococcus thermophilus, Bifidobacterium longum, Bifidobacterium breve, and Bifidobacterium infantis. # It has been shown that administration of 3 suppresses weight loss, reduces the disease activity index (DAI), and improves intestinal lesions in a mouse model of ulcerative colitis. Non-patent literature 1 describes that Lactobacillus delbrueckii LAB4 suppresses the excessive response of additional insulin secretion following the initial rise in blood glucose level immediately after sugar intake, thereby suppressing hypoglycemia. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Patent Publication No. 2015-205829 [Patent Document 2] Patent Publication No. 2015-205830 [Patent Document 3] Patent No. 6657084 (International Publication WO2015 / 199192) [Patent Document 4] Patent No. 6813974 (JP 2018-8901 A) [Patent Document 5] International Publication WO2020 / 218450 [Patent Document 6] Patent No. 5801802 (International Publication WO2011 / 063117) [Patent Document 7] International Publication WO2017 / 015407 [Patent Document 8] Patent Publication No. 2020-184906 [Patent Document 9] Special table 2021-524751 [Non-patent literature]

[0007] [Non-Patent Document 1] K. Nagata, C. Otsuki, S. Shiwa, M. Maekawa, T. Sugioka, Y. Ikeuchi, Hypothesis on the mechanism of action of Lactobacillus casei LAB4 and glucomannan-containing Lactobacillus casei LAB4 against blood glucose spikes, Comprehensive Medicine 20 (1) 3-12, 2021. Summary of the Invention [Problem to be solved by the invention]

[0008] An object of the present invention is to provide a novel composition for promoting ghrelin secretion. [Means for solving the problem]

[0009] The present invention relates to the following: <1> A composition for promoting ghrelin secretion, comprising bacteria belonging to Lactobacillus delbrueckii. <2> A composition comprising bacteria belonging to Lactobacillus delbrueckii for one or more effects selected from the group consisting of promotion of growth hormone secretion, inhibition of brown adipose tissue function, promotion of gastrointestinal motility, enhancement of gastric acid secretion, enhancement of pancreatic exocrine secretion, vasodilation, increase in cardiac contractility, and protection of myocardial cells. <3> At least one of the following effects is due to the promotion of ghrelin secretion: promotion of growth hormone secretion, inhibition of brown adipose tissue function, promotion of gastrointestinal motility, enhancement of gastric acid secretion, enhancement of pancreatic exocrine secretion, vasodilation, increase in cardiac contractility, and protection of myocardial cells. <2> The composition described in <4> The bacterium belonging to Lactobacillus delbrueckii belongs to any one of Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus delbrueckii ssp. delbrueckii, Lactobacillus delbrueckii ssp. indicus, Lactobacillus delbrueckii ssp. lactis, and Lactobacillus delbrueckii ssp. sunkii. <1> ~ <3> 13. The composition according to any one of claims 1 to 12. <5> A bacterium belonging to Lactobacillus delbrueckii has a 16S rRNA gene having 90% or more identity with a base sequence represented by any one of SEQ ID NOs: 1 to 5. <1> ~ <4> 13. The composition according to any one of claims 1 to 12. <6> Bacteria belonging to Lactobacillus delbrueckii ssp. bulgaricus were identified as Lactobacillus delbrueckii ssp. bulgaricus 2038, Lactobacillus delbrueckii ssp. bulgaricus OLL1255 (Accession Number: NITE BP-76), Lactobacillus delbrueckii ssp. bulgaricus OLL1171 (Accession Number: NITE BP-01569), Lactobacillus delbrueckii ssp. bulgaricus OLL1247 (Accession Number: NITE BP-01814), Lactobacillus delbrueckii ssp. bulgaricus OLL205013 (Accession Number: NITE BP-02411), and Lactobacillus delbrueckii OLL204989 (Accession Number: NITE BP-02412). BP-02874), Lactobacillus delbrueckii ssp. delbrueckii (JCM 1012T), Lactobacillus delbrueckii ssp. indicus (JCM 15610T), Lactobacillus delbrueckii ssp. lactis (JCM 1248T), and Lactobacillus delbrueckii ssp. sunkii (JCM 17838T), <1> ~ <5> 13. The composition according to any one of claims 1 to 12. <7> The bacteria belonging to Lactobacillus delbrueckii are viable bacteria. <1> ~ <6> 13. The composition according to any one of claims 1 to 12. <8> The composition is a fermented milk that does not contain bacteria belonging to Lactobacillus paragaceri. <1> ~ <7> 13. The composition according to any one of claims 1 to 12. <9> A bacterium belonging to Lactobacillus delbrueckii does not have a DNA having the sequence of SEQ ID NO: 6. <1> ~ <8> 13. The composition according to any one of claims 1 to 12. <10> A pharmaceutical composition comprising a bacterium belonging to the group Lactobacillus delbrueckii. <11> It is a hormone drug <10> The pharmaceutical composition described in

[0010] The present invention also relates to the following: [1] A composition for promoting ghrelin secretion, comprising bacteria belonging to the Lactobacillus delbrueckii species. [2] A composition comprising a bacterium belonging to Lactobacillus delbrueckii, for any one selected from the group consisting of promotion of growth hormone secretion, promotion of fat accumulation (suppression of weight loss), suppression of insulin secretion, suppression of brown adipose tissue function, promotion of gastrointestinal motility, enhancement of gastric acid secretion, enhancement of pancreatic exocrine secretion, vasodilation, increase in cardiac contractility, and protection of cardiomyocytes. [3] The composition described in [2], which promotes ghrelin secretion. [4] The composition according to any one of [1] to [3], wherein the bacterium belonging to Lactobacillus delbrueckii belongs to any one of Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus delbrueckii ssp. delbrueckii, Lactobacillus delbrueckii ssp. indicus, Lactobacillus delbrueckii ssp. lactis, and Lactobacillus delbrueckii ssp. sunkii. [5] Bacteria belonging to Lactobacillus delbrueckii ssp. bulgaricus include Lactobacillus delbrueckii ssp. bulgaricus 2038, Lactobacillus delbrueckii ssp. bulgaricus OLL1255 (Accession Number: NITE BP-76), Lactobacillus delbrueckii ssp. bulgaricus OLL1171 (Accession Number: NITE BP-01569), Lactobacillus delbrueckii ssp. bulgaricus OLL1247 (Accession Number: NITE BP-01814), Lactobacillus delbrueckii ssp. bulgaricus OLL205013 (Accession Number: NITE BP-02411), and Lactobacillus delbrueckii OLL204989 (Accession Number: NITE BP-02412). BP-02874), Lactobacillus delbrueckii ssp. delbrueckii (JCM 1012T), Lactobacillus delbrueckii ssp. indicus (JCM 15610T), Lactobacillus delbrueckii ssp. lactis (JCM 1248T), and Lactobacillus delbrueckii ssp. sunkii (JCM 17838T). [6] The composition according to any one of [1] to [5], wherein the bacterium belonging to Lactobacillus delbrueckii is a live bacterial cell. [7] The composition described in any one of [1] to [6], which is a fermented milk that does not contain bacteria belonging to Lactobacillus paragaceri. [8] The composition according to any one of [1] to [7], wherein the bacterium belonging to Lactobacillus delbrueckii does not have DNA consisting of the sequence of SEQ ID NO: 6.

[0011] The present invention also relates to the following: [9] A composition comprising bacteria belonging to Lactobacillus delbrueckii for use in a method for promoting ghrelin secretion. Use of bacteria belonging to Lactobacillus delbrueckii in the manufacture of a composition for promoting ghrelin secretion. A method or non-therapeutic method for promoting ghrelin secretion, comprising the step of administering to a subject a composition comprising bacteria belonging to Lactobacillus delbrueckii. Use or non-therapeutic use of a composition comprising bacteria belonging to Lactobacillus delbrueckii to promote ghrelin secretion.

[10] A composition containing a bacterium belonging to Lactobacillus delbrueckii for use in any method selected from the group consisting of promotion of growth hormone secretion, promotion of fat accumulation (suppression of weight loss), suppression of insulin secretion, inhibition of brown adipose tissue function, promotion of gastrointestinal motility, enhancement of gastric acid secretion, enhancement of exocrine pancreatic secretion, vasodilation, increased cardiac contractility, and protection of myocardial cells. Use of a bacterium belonging to Lactobacillus delbrueckii in the manufacture of a composition for any method selected from the group consisting of promotion of growth hormone secretion, promotion of fat accumulation (suppression of weight loss), suppression of insulin secretion, inhibition of brown adipose tissue function, promotion of gastrointestinal motility, enhancement of gastric acid secretion, enhancement of exocrine pancreatic secretion, vasodilation, increased cardiac contractility, and protection of myocardial cells. A method or non-therapeutic method for any one selected from the group consisting of promotion of growth hormone secretion, promotion of fat accumulation (suppression of weight loss), suppression of insulin secretion, suppression of brown adipose tissue function, promotion of gastrointestinal motility, enhancement of gastric acid secretion, enhancement of exocrine pancreatic secretion, vasodilation, increased cardiac contractility, and protection of myocardial cells, comprising a step of administering to a subject a composition containing bacteria belonging to Lactobacillus delbrueckii. Use or non-therapeutic use of a composition containing bacteria belonging to Lactobacillus delbrueckii for any one selected from the group consisting of promotion of growth hormone secretion, promotion of fat accumulation (suppression of weight loss), suppression of insulin secretion, suppression of brown adipose tissue function, promotion of gastrointestinal motility, enhancement of gastric acid secretion, enhancement of exocrine pancreatic secretion, vasodilation, increased cardiac contractility, and protection of myocardial cells.

[11] The composition, use, method or non-therapeutic method for manufacturing, or use or non-therapeutic use described in

[10] , wherein any of the effects selected from the group consisting of promotion of growth hormone secretion, promotion of fat accumulation (suppression of weight loss), suppression of insulin secretion, suppression of brown adipose tissue function, promotion of gastrointestinal motility, enhanced gastric acid secretion, enhanced pancreatic exocrine secretion, vasodilation, increased cardiac contractility, and protection of cardiomyocytes is due to promotion of ghrelin secretion.

[12] The composition, use, method or non-therapeutic method for production, or use or non-therapeutic use according to any one of [9] to

[11] , wherein the bacterium belonging to Lactobacillus delbrueckii belongs to any of Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus delbrueckii ssp. delbrueckii, Lactobacillus delbrueckii ssp. indicus, Lactobacillus delbrueckii ssp. lactis, and Lactobacillus delbrueckii ssp. sunkii.

[13] Bacteria belonging to Lactobacillus delbrueckii ssp. bulgaricus include Lactobacillus delbrueckii ssp. bulgaricus 2038, Lactobacillus delbrueckii ssp. bulgaricus OLL1255 (Accession Number: NITE BP-76), Lactobacillus delbrueckii ssp. bulgaricus OLL1171 (Accession Number: NITE BP-01569), Lactobacillus delbrueckii ssp. bulgaricus OLL1247 (Accession Number: NITE BP-01814), Lactobacillus delbrueckii ssp. bulgaricus OLL205013 (Accession Number: NITE BP-02411), and Lactobacillus delbrueckii OLL204989 (Accession Number: NITE BP-02412). BP-02874), Lactobacillus delbrueckii ssp. delbrueckii (JCM 1012T), Lactobacillus delbrueckii ssp. indicus (JCM 15610T), Lactobacillus delbrueckii ssp. lactis (JCM 1248T), and Lactobacillus delbrueckii ssp. sunkii (JCM 17838T).

[14] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of [9] to

[13] , wherein the bacterium belonging to Lactobacillus delbrueckii is a viable bacterium.

[15] The composition, use, method or non-therapeutic method, or use or non-therapeutic use described in any one of [9] to

[14] , wherein the composition is fermented milk not containing bacteria belonging to Lactobacillus paragaceri.

[16] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use described in any one of [1] to [7], wherein the bacterium belonging to Lactobacillus delbrueckii does not have DNA consisting of the sequence of SEQ ID NO: 6. Effect of the Invention

[0012] According to the present invention, it is possible to promote the secretion of ghrelin using highly safe ingredients. [Brief description of the drawings]

[0013] [Figure 1] Figure 1 shows the results of measuring the mRNA expression level of ghrelin in MKN45 cells incubated with each strain of Lactobacillus delbrueckii subsp. bulgaricus for 4 or 6 hours. Means ± SE, n=3, *p<0.05, **p<0.01 vs. control (by Dunnett test). [Diagram 2] Figure 2 shows the results of measuring the mRNA expression level of ghrelin when using Lactobacillus delbrueckii OLL204989, Lactobacillus delbrueckii ssp. delbrueckii JCM 1012T, Lactobacillus delbrueckii ssp. indicus JCM 15610T, Lactobacillus delbrueckii subsp. lactis JCM 1248T, or Lactobacillus delbrueckii ssp. sunkii JCM 17838T. Means ± SE, n=3, *p<0.05, **p<0.01 vs. control (by Dunnett test) DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The present invention will be described in detail below. In one aspect, the present invention relates to a composition comprising a specific bacterium. In one aspect, the present invention relates to a composition for promoting ghrelin secretion, etc., containing a specific bacterium as an active ingredient. Note that in the present invention, the term "any" means that the number and type are optional.

[0015] [Active ingredients] "Bacteria belonging to Lactobacillus delbrueckii" The composition of the present invention contains bacteria belonging to Lactobacillus delbrueckii (hereinafter, sometimes referred to as L. delbrueckii) as an active ingredient. In the composition, any bacteria belonging to L. delbrueckii can be used as the active ingredient, regardless of the specific subspecies.

[0016] Including an active ingredient means that the ingredient is used in the composition in an effective amount to exert the intended function, that the ingredient is specified in the label as contributing to the intended purpose, etc. In foods with functional claims, active ingredients are sometimes called functional ingredients (ingredients that contribute to specific health purposes (excluding those related to reducing the risk of disease)).

[0017] A strain taxonomically equivalent to a certain strain (hereinafter referred to as strain S) refers to, for example, any of the following: a strain belonging to the same genus as strain S, preferably belonging to the same species as strain S, the entire sequence or a characteristic part of the 16S rRNA gene of which (the V1 region, the V2 region, or the entire or part of the V1 and V2 regions, or a part including the V1 and V2 regions, etc.) has a sequence identity of 90% or more, preferably 95% or more, more preferably 98% or more, even more preferably 98.5% or more, even more preferably more than 98.7%, even more preferably 99% or more, even more preferably 100% with the sequence of strain S; A strain with the same mycological properties as strain S.

[0018] Regarding the criteria for determining the identity and difference of species based on the 16S rRNA gene sequence, those skilled in the art can refer to Stackebrandt E, Ebers J. Taxonomic parameters revisited: tarnished gold standards. Microbiol Today 2006;33:152-155.

[0019] (L. delbrueckii OLL204989) As an example of a strain belonging to L. delbrueckii, OLL204989 (Accession No.: NITE BP-02874) and strains taxonomically equivalent thereto are used. L. delbrueckii OLL204989 has been deposited internationally under the accession number NITE BP-02874 at the Patent Microorganisms Depositary, National Institute of Technology and Evaluation (2-5-8 Kazusa Kamatari, Kisarazu, Chiba, Japan) (Depositor: Meiji Co., Ltd., Date of deposit for international deposit based on the Budapest Treaty: February 5, 2019).

[0020] <Bacteria belonging to Lactobacillus delbrueckii ssp. bulgaricus> In one preferred embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention are bacteria belonging to Lactobacillus delbrueckii subsp. bulgaricus (hereinafter sometimes referred to as L. delbrueckii ssp. bulgaricus). L. delbrueckii ssp. bulgaricus is a lactic acid bacterium suitable for fermented milk, and the use thereof facilitates the production of fermented milk. In the composition, as long as it is a bacterium belonging to L. delbrueckii ssp. bulgaricus, various strains can be used as an active ingredient without being limited to a specific strain.

[0021] Examples of strains belonging to L. delbrueckii ssp. bulgaricus include 2038, OLL1255 (Accession No.: NITE BP-76), OLL1171 (Accession No.: NITE BP-01569), OLL1247 (Accession No.: NITE BP-01814), OLL205013 (Accession No.: NITE BP-02411), and taxonomically equivalent strains.

[0022] (L. delbrueckii ssp. bulgaricus 2038) L. delbrueckii ssp. bulgaricus 2038 can be isolated from Meiji Bulgaria Yogurt LB81 (Meiji Co., Ltd.) and is commercially available. It is also stored at Meiji Co., Ltd. (Meiji Innovation Center, 1-29-1 Shichikuni, Hachioji-shi, Tokyo). The 2038 strain can also be isolated, for example, by spreading a diluted solution of Meiji Bulgaria Yogurt LB81 (manufactured by Meiji Co., Ltd.) on a BCP-added agar medium, culturing it at 37°C for 48 hours, and then picking out rough colonies.

[0023] The mycological properties of 2038 are shown below. Morphology: Bacillus; does not produce gas from glucose; GC content 49.7%; gram positive; catalase negative; lactic acid optical rotation: D type; does not grow at 15°C; produces acid by assimilating glucose, lactose, etc.; does not form spores; non-motile.

[0024] The sequence of the 16S rRNA gene of 2038 (corresponding to the 27th to 520th positions in the full-length sequence of the rRNA gene) is shown in SEQ ID NO:1 in the sequence listing. 16S rRNA gene, Lactobacillus delbrueckii ssp.bulgaricus 2038(SEQ ID NO:1) gctggcggcgtgcctaatacatgcaagtcgagcgagctgaattcaaagatyccttcgggrtgatttgttggacgctagcggcggatgggtgagtaacacgtgggcaatctgccctaaagactggg ataccacttggaaacaggtgctaataccggataacaacatgaatcgcatgattcaagtttgaaaggcggcgyaagctgtcactttaggatgagcccgcggcgcattagctagttggtggggtaaag gcctaccaaggcaatgatgcgtagccgagttgagagactgatcggccacattgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatggacgcaagtctgat ggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaagctctgttgttggtgaagaaggatagaggcagtaactggtctttatttgacggtaatcaaccagaaagtcacggctaactacgt

[0025] (L. delbrueckii ssp. bulgaricus OLL1255) L. delbrueckii ssp. bulgaricus OLL1255 (accession number: NITE BP-76) has been internationally deposited at the National Institute of Technology and Evaluation, Patent Organism Depositary Center (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture) under the accession number NITE BP-76 (depositor: Meiji Co., Ltd., date of domestic deposit: February 10, 2005, date of receipt of request for transfer to international deposit under the Budapest Treaty: April 1, 2009).

[0026] The mycological properties of OLL1255 are shown below. Morphology: rod-shaped, no gas production, gram positive, catalase negative, facultative anaerobic, no nitrate reduction, no indole production, no hydrogen sulfide production, lactic acid optical rotation: D type, no growth at 15°C, no spore formation, no motility. Glycolytic: Arabinose -, Galactose -, Glucose +, Salicin -, Sucrose -, Cellobios -, Starch -, Maltose -, Mannitol -, Mannose +, Lactose +, Raffinose -, Ribose -.

[0027] The sequence of the 16S rRNA gene of OLL1255 (corresponding to positions 27 to 520 in the full-length sequence of the rRNA gene) is shown in SEQ ID NO:2 in the sequence listing. 16S rRNA gene, Lactobacillus delbrueckii ssp.bulgaricus OLL1255(SEQ ID NO:2) gctggcggcgtgcctaatacatgcaagtcgagcgagctgaattcaaagatyccttcgggrtgatttgttggacgctagcggcggatgggtgagtaacacgtgggcaatctgccctaaagactggg ataccacttggaaacaggtgctaataccggataacaacatgaatcgcatgattcaagtttgaaaggcggcgyaagctgtcactttaggatgagcccgcggcgcattagctagttggtggggtaaag gcctaccaaggcaatgatgcgtagccgagttgagagactgatcggccacattgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatggacgcaagtctgat ggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaagctctgttgttggtgaagaaggatagaggcagtaactggtctttatttgacggtaatcaaccagaaagtcacggctaactacgt

[0028] (L. delbrueckii ssp. bulgaricus OLL1171) L. delbrueckii ssp. bulgaricus OLL1171 (Accession Number: NITE BP-01569) has been internationally deposited at the National Institute of Technology and Evaluation, Patent Organism Depositary Center (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture) under the accession number NITE BP-01569 (Depositor: Meiji Co., Ltd.; Date of international deposit under the Budapest Treaty: March 13, 2013).

[0029] The sequence of the 16S rRNA gene of OLL1171 (corresponding to positions 27 to 520 in the full-length sequence of the rRNA gene) is shown in SEQ ID NO:3 in the sequence listing. 16S rRNA gene, Lactobacillus delbrueckii ssp.bulgaricusOLL1171(SEQ ID NO:3) gctggcggcgtgcctaatacatgcaagtcgagcgagctgaattcaaagatyccttcgggrtgatttgttggacgctagcggcggatgggtgagtaacacgtgggcaatctgccctaaagactggg ataccacttggaaacaggtgctaataccggataacaacatgaatcgcatgattcaagtttgaaaggcggcgyaagctgtcactttaggatgagcccgcggcgcattagctagttggtggggtaaag gcctaccaaggcaatgatgcgtagccgagttgagagactgatcggccacattgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatggacgcaagtctgat ggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaagctctgttgttggtgaagaaggatagaggcagtaactggtctttatttgacggtaatcaaccagaaagtcacggctaactacgt

[0030] (L. delbrueckii ssp. bulgaricus OLL1247) L. delbrueckii ssp. bulgaricus OLL1247 (Accession Number: NITE BP-01814) has been internationally deposited at the National Institute of Technology and Evaluation, Patent Organism Depositary Center (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture) under the accession number NITE BP-01814 (Depositor: Meiji Co., Ltd., Date of international deposit under the Budapest Treaty: March 6, 2014).

[0031] The sequence of the 16S rRNA gene of OLL1247 (corresponding to positions 27 to 520 in the full-length sequence of the rRNA gene) is shown in SEQ ID NO:4 in the sequence listing. 16S rRNA gene, Lactobacillus delbrueckii ssp.bulgaricusOLL1247(SEQ ID NO:4) gctggcggcgtgcctaatacatgcaagtcgagcgagctgaattcaaagatyccttcgggrtgatttgttggacgctagcggcggatgggtgagtaacacgtgggcaatctgccctaaagactggg ataccacttggaaacaggtgctaataccggataacaacatgaatcgcatgattcaagtttgaaaggcggcgyaagctgtcactttaggatgagcccgcggcgcattagctagttggtggggtaaag gcctaccaaggcaatgatgcgtagccgagttgagagactgatcggccacattgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatggacgcaagtctgat ggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaagctctgttgttggtgaagaaggatagaggcagtaactggtctttatttgacggtaatcaaccagaaagtcacggctaactacgt

[0032] (L. delbrueckii ssp. bulgaricus OLL205013) L. delbrueckii ssp. bulgaricus OLL205013 (Deposit No.: NITE BP-02411) is internationally deposited under the deposit number NITE BP-02411 at the Patent Biological Depositary, National Institute of Technology and Evaluation (2-5-8 Kazusa-Kamatari, Kisarazu, Chiba, Japan) (Depositor: Meiji Co., Ltd.; Date of international deposit based on the Budapest Treaty: February 13, 2017).

[0033] The sequence of the 16S rRNA gene of OLL205013 (corresponding to positions 27 to 520 in the full-length sequence of the rRNA gene) is shown as SEQ ID NO:5 in the Sequence Listing. 16S rRNA gene, Lactobacillus delbrueckii ssp.bulgaricus OLL205013(SEQ ID NO:5) gctggcggcgtgcctaatacatgcaagtcgagcgagctgaattcaaagatyccttcgggrtgatttgttggacgctagcggcggatgggtgagtaacacgtgggcaatctgccctaaagactgggataccacttggaaacaggtgctaataccggataacaacatgaatcgcatgattcaagtttgaaaggcggcgyaagctgtcactttaggatgagcccgcggcgcattagctagttggtggggtaaaggcctaccaaggcaatgatgcgtagccgagttgagagactgatcggccacattgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatggacgcaagtctgatggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaagctctgttgttggtgaagaaggatagaggcagtaactggtctttatttgacggtaatcaaccagaaagtcacggctaactacgt

[0034] <Bacteria belonging to Lactobacillus delbrueckii ssp. delbrueckii> In one preferred embodiment, the bacterium belonging to L. delbrueckii in the composition of the present invention is a bacterium belonging to Lactobacillus delbrueckii ssp. delbrueckii (hereinafter, may also be referred to as L. delbrueckii ssp. delbrueckii). In the composition, as long as it is a bacterium belonging to L. delbrueckii ssp. delbrueckii, various ones can be used as active ingredients without being limited to a specific strain.

[0035] (L. delbrueckii ssp. delbrueckii JCM 1012 T ) Examples of strains belonging to L. delbrueckii ssp. delbrueckii include JCM 1012 T and strains taxonomically equivalent thereto are used. L. delbrueckii ssp. delbrueckii JCM 1012 T can be obtained from the Microbial Material Development Unit of the RIKEN BioResource Center (RIKEN BRC) (Japan Collection of Microorganisms) (RIKEN BRC-JCM, Japan) under the JCM number JCM 1012 T .

[0036] <Bacterium belonging to Lactobacillus delbrueckii ssp. indicus> In one preferred embodiment, the bacterium belonging to L. delbrueckii in the composition of the present invention is a bacterium belonging to Lactobacillus delbrueckii ssp. indicus (hereinafter, may also be referred to as L. delbrueckii ssp. indicus). In the composition, as long as it is a bacterium belonging to L. delbrueckii ssp. indicus, various ones can be used as active ingredients without being limited to a specific strain.

[0037] (L. delbrueckii ssp. indicus JCM 15610 T ) As an example of a strain belonging to L. delbrueckii ssp. indicus, JCM 15610 T and strains taxonomically equivalent thereto are used. L. delbrueckii ssp. indicus JCM 15610 T is available from the Microbial Materials Development Unit, RIKEN BioResource Center (Japan Collection of Microorganisms) (RIKEN BRC-JCM, Japan) under the JCM number JCM 15610 T of.

[0038] <Bacteria belonging to Lactobacillus delbrueckii ssp. lactis> In one preferred embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention are bacteria belonging to Lactobacillus delbrueckii subsp. lactis (hereinafter sometimes referred to as L. delbrueckii ssp. lactis). In the composition, any bacteria belonging to L. delbrueckii ssp. lactis can be used as an active ingredient without being limited to a specific strain.

[0039] (L. delbrueckii ssp. lactis JCM 1248 T ) As an example of a strain belonging to L. delbrueckii ssp. lactis, JCM 1248 T and strains taxonomically equivalent thereto are used. L. delbrueckii ssp. lactis JCM 1248 Tis available from the RIKEN BioResource Center (RIKEN BRC), Japan Collection of Microorganisms (RIKEN BRC-JCM, Japan) under JCM accession number JCM 1248 T and can be obtained under

[0040] <Bacteria belonging to Lactobacillus delbrueckii ssp. sunkii> In one preferred embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention are bacteria belonging to Lactobacillus delbrueckii subsp. sunkii (hereinafter sometimes referred to as L. delbrueckii ssp. sunkii). In the composition, as long as the bacteria belong to L. delbrueckii ssp. sunkii, various strains can be used as active ingredients without being limited to a specific strain.

[0041] (L. delbrueckii ssp. sunkii JCM 17838 T ) Examples of strains belonging to L. delbrueckii ssp. sunkii include JCM 17838 T and strains taxonomically equivalent thereto are used. L. delbrueckii ssp. sunkii JCM 17838 T is available from the RIKEN BioResource Center (RIKEN BRC), Japan Collection of Microorganisms (RIKEN BRC-JCM, Japan) under JCM accession number JCM 17838 T and can be obtained under

[0042] As described above, in one preferred embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention belong to any one of L. delbrueckii ssp., L. delbrueckii ssp. bulgaricus, L. delbrueckii ssp. delbrueckii, L. delbrueckii ssp. indicus, L. delbrueckii ssp. lactis, and L. delbrueckii ssp. sunkii. In another embodiment, the bacteria belonging to L. delbrueckii do not include bacteria belonging to Lactobacillus delbrueckii ssp. bulgaricus. In yet another embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention belong to any one of L. delbrueckii ssp. delbrueckii, L. delbrueckii ssp. indicus, L. delbrueckii ssp. lactis, and L. delbrueckii ssp. sunkii.

[0043] In addition, the bacteria belonging to L. delbrueckii in the composition of the present invention include L. delbrueckii ssp. bulgaricus 2038, L. delbrueckii ssp. bulgaricus OLL1255 (Accession Number: NITE BP-76), L. delbrueckii ssp. bulgaricus OLL1171 (Accession Number: NITE BP-01569), L. delbrueckii ssp. bulgaricus OLL1247 (Accession Number: NITE BP-01814), L. delbrueckii ssp. bulgaricus OLL205013 (Accession Number: NITE BP-02411), L. delbrueckii OLL204989 (Accession Number: NITE BP-02874), L. delbrueckii ssp. delbrueckii (JCM 1012T), L. In another embodiment, the bacterium belonging to L. delbrueckii in the composition of the present invention does not include L. delbrueckii LAB4. In yet another embodiment, the bacterium belonging to L. delbrueckii in the composition of the present invention is any one or more selected from the group consisting of L. delbrueckii ssp. delbrueckii (JCM 1012T), L. delbrueckii ssp. indicus (JCM 15610T), L. delbrueckii ssp. lactis (JCM 1248T), and L. delbrueckii ssp. sunkii (JCM 17838T).

[0044] In another aspect, the bacterium belonging to L. delbrueckii in the composition of the present invention has a 16S rRNA gene having an identity of 90% or more, preferably 95% or more, more preferably 98% or more, still more preferably 98.5% or more, still more preferably 98.7% or more, particularly preferably 99% or more, preferably 99.5% or more with the nucleotide sequence represented by any one of SEQ ID NOs: 1 to 5.

[0045] A composition containing any one of the above strains as an active ingredient is preferable in that the intended effect is particularly excellent.

[0046] <DNA encoding hypothetical protein gene> According to the study by the present inventors, as one of the common characteristics of the strains showing a ghrelin secretion promoting action under the conditions of this example, it was found that they do not have the DNA of the sequence described below as SEQ ID NO: 6 (DNA encoding hypothetical protein gene). The amino acid sequence of the protein (hypothetical protein) corresponding to this DNA is described as SEQ ID NO: 7 in the sequence listing. Therefore, in one preferred aspect, the composition may contain, as an active ingredient, any bacterium belonging to Lactobacillus delbrueckii that does not have the DNA of the following (i). In other words, in this aspect, the bacterium belonging to Lactobacillus delbrueckii contained in the composition does not have the DNA of (i) or does not contain a strain having the DNA of (i). In another aspect, the composition may contain, as an active ingredient, any bacterium belonging to Lactobacillus delbrueckii that does not have the DNA of the following (ii). In still another aspect, the composition may contain, as an active ingredient, any bacterium belonging to Lactobacillus delbrueckii that does not have the DNA of the following (iii). (i) DNA consisting of the sequence of SEQ ID NO: 6 (ii) DNA consisting of a sequence having at least 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more identity to SEQ ID NO: 6 (iii) DNA consisting of a sequence in which 1, 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 1 to 11, 1 to 12, 1 to 13, 1 to 14, 1 to 15, 1 to 16, 1 to 17, 1 to 18, 1 to 19, 1 to 20, or 1 to 21 bases are substituted, deleted, or added in SEQ ID NO: 6.

[0047] In yet another preferred embodiment, the composition may contain, as an active ingredient, any bacterium belonging to Lactobacillus delbrueckii that does not have the protein (iv) below. In other words, in this embodiment, the bacterium belonging to Lactobacillus delbrueckii contained in the composition does not have the protein (iv) or does not include a strain having the protein (iv). In another embodiment, the composition may contain, as an active ingredient, any bacterium belonging to Lactobacillus delbrueckii that does not have the protein (v) below. In yet another embodiment, the composition may contain, as an active ingredient, any bacterium belonging to Lactobacillus delbrueckii that does not have the protein (vi) below. (iv) a protein consisting of the sequence of SEQ ID NO: 7 (v) A protein encoded by the DNA of (ii) above. (vi) A protein encoded by the DNA of (iii) above.

[0048] In yet another embodiment, the composition may contain any bacterium that does not have DNA (which may include DNA of SEQ ID NO: 6) encoding a protein consisting of the sequence of SEQ ID NO: 7. In other words, in this embodiment, the bacterium belonging to Lactobacillus delbrueckii contained in the composition does not have DNA encoding a protein consisting of the sequence of SEQ ID NO: 7, or does not include a strain having DNA encoding a protein consisting of the sequence of SEQ ID NO: 7. In another preferred embodiment, the composition may contain any bacterium belonging to Lactobacillus delbrueckii ssp. bulgaricus as an active ingredient, which does not have DNA consisting of the sequence of SEQ ID NO: 6. In another preferred embodiment, the composition may contain any bacterium belonging to Lactobacillus delbrueckii ssp. bulgaricus as an active ingredient, which does not have a protein consisting of the sequence of SEQ ID NO: 7.

[0049] DNA encoding hypothetical protein gene (SEQ ID NO:6) atggttgatgagatgtaccctgaagccgagaagatcattcttgtcatagacaatcttaatacgcacaagccatcctcactatataaggctttcaaagcacgtcgaatc atcaagaagcttgagattcactatactcccaagcacggcagctggttagatatcgctgaaattgatgatgaccagacagtgtctttctcgaagaatcccggatattga

[0050] hypothetical protein (SEQ ID NO:7) MVDEMYPEAEKIILVIDNLNTHKPSSLYKAFKARRIIKKLEIHYTPKHGSWLDIAEIDDDQTVSFSKNPGY

[0051] The composition may contain lactic acid bacteria other than bacteria belonging to L. delbrueckii, so long as the intended function is not impaired. The composition may also contain microorganisms such as yeast other than the intended lactic acid bacteria. In a preferred embodiment, the composition contains one or more types of lactic acid bacteria, but does not contain other microorganisms, such as yeast. In another preferred embodiment, the composition does not contain either or both of Lactobacillus gasseri and Lactobacillus paragasseri. In another preferred embodiment, the composition does not contain Streptococcus thermophilus. In another preferred embodiment, the composition does not contain L. delbrueckii ssp. bulgaricus OLL1073R-1. In another preferred embodiment, the composition does not contain lactic acid bacteria other than bacteria belonging to L. delbrueckii. In another preferred embodiment, the composition does not contain lactic acid bacteria other than bacteria belonging to L. delbrueckii ssp. bulgaricus. In another preferred embodiment, the composition does not contain any component that promotes ghrelin secretion other than bacteria belonging to L. delbrueckii.In another preferred embodiment, the composition does not contain any component that promotes ghrelin secretion other than bacteria belonging to L. delbrueckii ssp.

[0052] The lactic acid bacteria (effective bacteria) that are the active ingredients contained in the composition can be produced by culturing. The culture conditions are not particularly limited as long as the desired effect is achieved.

[0053] In the composition of the present invention, the effective bacteria may be contained in any state as long as it can exert the intended effect. For example, the effective bacteria may be the effective bacteria itself, or may be a culture of the effective bacteria (consisting of the effective bacteria and culture supernatant). The bacteria may be in a living state (living bacteria) or in a dead state (dead bacteria) as long as it can exert the intended effect.

[0054] The effective bacteria may be contained in the composition as killed bacteria. The killed bacteria can be obtained by sterilizing the effective bacteria. The sterilization is not particularly limited as long as it can exert the intended effect, and can be performed by heating, a germicidal lamp (UV), ozone, a drug, high osmotic pressure, etc.

[0055] The killed bacteria are preferably heat-killed bacteria obtained by heat-treating live bacteria. The heat treatment for obtaining heat-killed bacteria is not particularly limited as long as the desired effect is achieved, and is carried out at a temperature and time sufficient to kill the effective bacteria used. Although such conditions depend on the effective bacteria used, the heat treatment temperature is, for example, 55°C or higher, preferably 60°C or higher, more preferably 65°C or higher, and even more preferably 70°C or higher, and may be 80°C or higher, or may be 90°C or higher. The upper limit of the heat treatment temperature can be appropriately set, for example, 121°C or lower, 100°C or lower, 90°C or lower, or 80°C or lower. Depending on the heat treatment temperature, the heat treatment time can be 1 minute or more, 3 minutes or more, 10 minutes or more, 15 minutes or more, 30 minutes or more, or 45 minutes or more. The upper limit of the heat treatment time can be, for example, 120 minutes or less, 100 minutes or less, 90 minutes or less, or 80 minutes or less.

[0056] The live bacterial cells may be obtained by culturing the live bacterial cells by a known method, and may be in the form of a culture solution, a concentrate of the culture solution, a frozen product, or a dried product. Alternatively, the culture solution may be subjected to a process such as centrifugation to recover only the bacterial cells, or the recovered bacterial cells may be suspended in a buffer. Alternatively, the live bacterial cells may be added (inoculated) to a food material such as milk (animal milk) that is the raw material for fermented foods, to increase the amount of the live bacterial cells inherent in the composition.

[0057] The effective bacteria to be contained in the composition, regardless of whether they are live or dead bacteria, can be prepared in the form of a dry substance, a suspension, a paste, a gel, or the like.

[0058] [Application] (function) In one aspect, the composition can be used to promote ghrelin secretion. Ghrelin is a peptide hormone secreted primarily in the stomach. The composition can also be used to aid, support, improve, or maintain ghrelin secretion. The composition can also be used to alleviate reduced ghrelin secretion, reduce the risk of reduced ghrelin secretion, or help improve ghrelin secretion.

[0059] Ghrelin is known to have actions such as growth hormone secretion promotion (Patent Documents 1 and 2), brown adipose tissue function inhibition, gastrointestinal motility promotion, gastric acid secretion promotion, pancreatic exocrine secretion promotion, vasodilation, cardiac contractile force increase, and cardiomyocyte protection (Patent Document 3). Therefore, in one embodiment, the composition can be used for any one or more selected from the group consisting of growth hormone secretion promotion, brown adipose tissue function inhibition, gastrointestinal motility promotion (or to assist digestion), gastric acid secretion promotion, pancreatic exocrine secretion promotion, vasodilation, cardiac contractile force increase, and cardiomyocyte protection. Alternatively, the composition can be used to regulate or help regulate any one or more selected from the group consisting of growth hormone secretion promotion, brown adipose tissue function inhibition, gastrointestinal motility promotion, gastric acid secretion promotion, pancreatic exocrine secretion promotion, vasodilation, cardiac contractile force increase, and cardiomyocyte protection.

[0060] In addition to the above-mentioned actions, ghrelin is known to have the actions of promoting fat accumulation (suppressing weight loss) and suppressing insulin secretion (Patent Document 3). Therefore, in one embodiment, the composition can be used for any one or more selected from the group consisting of promoting growth hormone secretion, promoting fat accumulation (suppressing weight loss), suppressing insulin secretion, suppressing the function of brown adipose tissue, promoting gastrointestinal motility (or aiding digestion), enhancing gastric acid secretion, enhancing exocrine pancreatic secretion, vasodilation, increasing cardiac contractility, and protecting myocardial cells. Alternatively, the composition can be used to regulate or help regulate any one or more selected from the group consisting of promoting growth hormone secretion, promoting fat accumulation (suppressing weight loss), suppressing insulin secretion, suppressing the function of brown adipose tissue, promoting gastrointestinal motility, enhancing gastric acid secretion, enhancing exocrine pancreatic secretion, vasodilation, increasing cardiac contractility, and protecting myocardial cells.

[0061] In addition, the uses of the composition of the present invention may exclude the following: autonomic nerve regulation, sympathetic nerve suppression, amelioration of sensitivity to chills, amelioration of stiffness, amelioration of gastrointestinal disorders, amelioration of worsening depression, amelioration of worsening dizziness, amelioration of increased heart rate, amelioration of increased blood pressure, amelioration of worsening chronic pain, amelioration of fatigue, amelioration of generalized lethargy, amelioration of generalized malaise, amelioration of subjective stress, amelioration of daytime sleepiness, amelioration of irritability, amelioration of loss of appetite, amelioration of decreased thinking ability, amelioration of thirst, amelioration of loss of refreshment, promotion of fat accumulation (inhibition of weight loss), inhibition of weight loss associated with ulcerative colitis, inhibition of insulin secretion, and inhibition of excessive insulin secretion after sugar ingestion.

[0062] Those skilled in the art can confirm whether a certain component promotes ghrelin secretion by various methods. For example, the component can be added to human stomach cells such as MKN45 cells (a cell line derived from human stomach cancer) and the expression level of the ghrelin gene in the cells after an appropriate period of time has elapsed, thereby making an evaluation. Specifically, the expression level of the ghrelin gene can be measured as the expression level of mRNA. It is preferable to correct for the expression level of a constitutively expressed gene, such as β-actin.

[0063] (subject) In one embodiment, the composition is suitable for administration to a subject in need of or desirably promoting secretion of ghrelin. In one embodiment, the composition is suitable for administration to a subject with reduced secretion of ghrelin. In another embodiment, the composition is suitable for administration to a subject in need of or desirably performing any one or more selected from the group consisting of promotion of growth hormone secretion, promotion of fat accumulation (suppression of weight loss), suppression of insulin secretion, suppression of brown adipose tissue function, promotion of gastrointestinal motility, enhancement of gastric acid secretion, enhancement of exocrine pancreatic secretion, vasodilation, increase in cardiac contractility, and protection of myocardial cells. The judgment of the disease or condition in the subject includes the judgment of a doctor, nurse, pharmacist, etc., lifestyle habits, eating habits, output results of a questionnaire on subjective symptoms, and the subject's own judgment based on subjective symptoms. The subject may be healthy (e.g., a person with a healthy stomach, no disease to be treated, and no administration of a drug for treatment, such as a chemotherapy agent). In addition, the following people may be excluded from the subjects who are to take the composition of the present invention: patients with autonomic nervous system disorders, people with poor circulation, people with stiffness, people with depression, people with dizziness, people with symptoms of summer fatigue, and patients with ulcerative colitis.

[0064] Such subjects include adults (15 years of age or older), middle-aged and elderly people, the elderly (65 years of age or older (as defined by the World Health Organization (WHO))), those undergoing or recovering from illness, pregnant women, postpartum women, infants, and children. Furthermore, since the active ingredient of the composition of the present invention is a bacterium that has a long history of consumption, it is suitable for long-term ingestion, and therefore is particularly suitable for subjects who wish to promote the secretion of ghrelin in their daily lives.

[0065] [Composition] (Food compositions, pharmaceutical compositions) The composition of the present invention can be a food composition or a pharmaceutical composition. The food and pharmaceutical products include not only those for humans but also those for animals other than humans, unless otherwise specified. The food includes general foods, functional foods, and nutritional compositions, as well as therapeutic foods (those that serve the purpose of treatment. Foods are prepared based on a menu prepared by a nutritionist or the like according to a doctor's dietary prescription), dietary therapy foods, ingredient-adjusted foods, nursing care foods, foods for medical treatment, and precision nutrition (individualized nutrition, appropriate meals (nutrition) proposed according to an individual's constitution). The food includes not only solid foods but also liquid foods, such as beverages, energy drinks, liquid foods, and soups, unless otherwise specified. Functional foods refer to foods that can provide a certain functionality to the living body, and include, for example, foods for specified health uses (including conditionally designated foods for specified health uses), foods with functional claims, health functional foods including foods with nutritional functions, foods for special dietary uses, dietary supplements, health supplements, dietary supplements, food supplements, medical foods (following the definition of the U.S. Food and Drug Administration (FDA)), supplements (for example, tablets, coated tablets, sugar-coated tablets, capsules, liquids, and other dosage forms), beauty foods (for example, diet foods), and other health foods in general. In addition, in the present invention, "functional foods" includes health foods to which a health claim based on the food standards of Codex (the Joint FAO / WHO Food Standards Commission) is applied. Food supplements are foods that supplement normal meals and are concentrated with nutrients or other substances that have nutritional or physiological effects, either alone or in combination, and are labeled as "Food Supplement" on the product. Dietary supplements are products (other than tobacco) intended to supplement the diet, that contain one or more of the targeted ingredients, and that are labeled as a dietary supplement.

[0066] Unless otherwise specified, pharmaceuticals include those that fall under the category of medical drugs, prescription drugs, over-the-counter drugs, and quasi-drugs as defined in the Act on Ensuring Quality, Efficacy, and Safety of Pharmaceuticals, Medical Devices, etc.

[0067] The compositions of the present invention may exclude: An agent containing a protein hydrolysate and a fermented milk protein as proteins; a medium-chain fatty acid as lipids; and a carbohydrate, wherein the fermented milk protein is derived from fermented milk obtained by fermenting skim milk using Lactobacillus Bulgaricus, Streptococcus Thermophilus, or a combination thereof; ·A composition comprising Lactobacillus casei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Bifidobacterium longum, Bifidobacterium breve, and Bifidobacterium infantis.

[0068] (Route of Administration) The composition of the present invention may be administered parenterally, for example, via a tube (gastrostomy, enterostomy), intranasally, or orally, but is preferably administered orally.

[0069] (Active ingredient content / dosage) The content of the effective bacteria in the composition of the present invention may be any amount that exerts the desired effect. The content of the effective bacteria in the composition may be appropriately determined taking into consideration various factors such as the age, weight, and symptoms of the subject to be ingested or administered. One unit of the composition may be the amount to be ingested or administered per day or once (which may be administered or taken once a day, or multiple times a day, for example, three times a day).

[0070] In one embodiment, the content of effective bacteria per unit of the composition is, for example, 1×10 5 Can be 1×10 or more 6 Can be 1×10 or more 7Can be 1×10 or more 8 More than 5×10 8 pcs or more, 5.7×10 8 pcs or more, 6×10 8 7×10 pieces or more 8 pcs or more, 8×10 8 pcs or more, 9×10 8 pcs or more, 1×10 9 More than 5×10 9 It can be more than 1×10 10 It is preferable to have more than 1×10 11 More preferably, 2×10 11 The upper limit can be set appropriately, and whatever the lower limit, it is preferably set to, for example, 1×10 14 It may be less than 1×10 13 It may be less than 1×10 12 It may be less than one.

[0071] In one embodiment, the effective bacterial intake is 1×10 5 Can be more than 1×10 pieces / day 6 Can be more than 1×10 pieces / day 7 Can be more than 1×10 pieces / day 8 pcs / day or more, 5×10 8 pcs / day or more, 5.7×10 8 pcs / day or more, 6×10 8 pcs / day or more, 7×10 8 pcs / day or more, 8×10 8 pcs / day or more, 9×10 8 pcs / day or more, 1×10 9 pcs / day or more, 5×10 9 1 x 10 pieces / day or more is acceptable 10 It is preferable to have more than 1.3 × 10 10 More preferably, it should be 2×10 10 pcs / day or more, 1×10 11 pcs / day or more, 2×10 11 It is more preferable that the number of cells / day is 1×10 or more. The upper limit can be set appropriately. Whatever the lower limit, the number of cells / day is 1×10 or more. 14It is acceptable to set the number of pieces per day or less, and 1×10 13 It is acceptable to set the number of pieces per day or less, and 1×10 12 It may be less than one per day.

[0072] In addition, commercially available yogurt contains 1×10 7 / g or more of L. delbrueckii ssp. bulgaricus, with each product containing 1 × 10 9 It is calculated that the specimens contain at least one L. delbrueckii ssp. bulgaricus.

[0073] In addition, when the content of effective bacteria is as described above, in any case, one unit can be, for example, 0.5g or more, 1g or more, 5g or more, 10g or more, 20g or more, or 30g or more. When the composition is in the form of a food such as fermented milk, one unit can be an amount that is easy to take in one time as a food, for example, 50g or more, 60g or more, 70g or more, 80g or more, 90g or more, or 100g or more. The upper limit can be appropriately set, and in any case, the lower limit can be 500g or less, 400g or less, 300g or less, 200g or less, 150g or less, or 125g or less.

[0074] The composition contains effective bacteria with a long history of consumption as an active ingredient, and therefore the composition of the present invention may be taken repeatedly or for a long period of time, for example, for 3 days or more, preferably 1 week or more, more preferably 15 days or more, further preferably 3 weeks or more, 4 weeks or more, or 1 month or more.

[0075] (Other ingredients, additives) The composition of the present invention may contain other active ingredients or nutritional ingredients that are acceptable as foods or pharmaceuticals. Examples of such ingredients include amino acids (e.g., lysine, arginine, glycine, alanine, glutamic acid, leucine, isoleucine, valine), carbohydrates (glucose, sucrose, fructose, maltose, trehalose, erythritol, maltitol, palatinose, xylitol, dextrin), electrolytes (e.g., sodium, potassium, calcium, magnesium), vitamins (e.g., vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, biotin, folic acid, pantothenic acid, and nicotinic acids), minerals (e.g., copper, zinc, iron, cobalt, manganese), antibiotics, dietary fiber, proteins, lipids, etc.

[0076] The composition may further contain additives that are acceptable for use as food or medicine. Examples of such additives include inert carriers (solid or liquid carriers), excipients, surfactants, binders, disintegrants, lubricants, solubilizers, suspending agents, coating agents, colorants, preservatives, buffers, pH adjusters, emulsifiers, stabilizers, sweeteners, antioxidants, flavors, acidulants, and natural products. More specifically, these include water, other aqueous solvents, pharma- ceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymers, sodium alginate, water-soluble dextran, water-soluble dextrin, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerin, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, sucralose, stevia, aspartame, acesulfame potassium, citric acid, lactic acid, malic acid, tartaric acid, phosphoric acid, acetic acid, fruit juice, vegetable juice, and the like.

[0077] (Dosage form / shape) The food composition of the present invention may be prepared in any form, such as solid, liquid, mixture, suspension, powder, granule, paste, jelly, gel, capsule, etc. The food composition of the present invention may be in any form, such as dairy products, supplements, confectionery, beverages, drinks, seasonings, processed foods, side dishes, soups, etc. More specifically, the composition of the present invention may be in the form of fermented milk, lactic acid bacteria beverage, dairy beverage, milk drink, soft drink, ice cream, tablet, cheese, bread, biscuit, cracker, pizza crust, modified milk powder, liquid food, food for sick people, nutritional food, frozen food, processed food, etc., and may be in the form of granule, powder, paste, thick liquid, etc., to be mixed with beverages or foods for ingestion. Fermented milk is fermented milk and lactic acid bacteria beverage defined in the "Ministerial Ordinance on the Compositional Standards of Milk and Dairy Products (hereinafter abbreviated as "Milk, etc. Ordinance"). Fermented milk, as defined in the Milk and Milk Products Ordinance, is milk or milk containing an equivalent or greater amount of non-fat milk solids that has been fermented with lactic acid bacteria or yeast to form a paste or liquid, or these products that have been frozen.

[0078] The pharmaceutical composition of the present invention can be in any dosage form suitable for oral administration, including solid preparations such as tablets, granules, powders, pills, and capsules; liquid preparations such as solutions, suspensions, and syrups; gels, and aerosols.

[0079] In one embodiment, the pharmaceutical composition of the present invention is a hormone agent. In another embodiment, the pharmaceutical composition of the present invention is a hormone secretagogue.

[0080] (others) In the production of the composition of the present invention, the stage of blending the effective bacteria can be appropriately selected. The stage of blending is not particularly limited as long as the properties of the effective bacteria are not significantly impaired. For example, the composition can be produced as fermented milk containing a sufficient amount of effective bacteria by mixing milk, a dairy product, and water, homogenizing as necessary, sterilizing, inoculating with the effective bacteria, and fermenting. The milk and dairy products used as raw materials can be appropriately selected according to the effective bacteria used for fermentation. In a preferred embodiment, the milk and dairy products used as raw materials include those derived from animals. In addition, the milk and dairy products used as raw materials do not have to include plant-based milk, such as soy milk, almond milk, oat milk, coconut milk, rice milk, and hemp milk.

[0081] The composition may be labeled with the intended use (use), and may also be labeled with a recommendation for ingestion to a specific target. In addition, the beginning of each statement may be appropriately labeled with a period such as "temporary" or "long-term". The specific target may be, for example, a target described in the above (target) section. Labeling may be direct or indirect. Examples of direct labeling include descriptions on tangible objects such as the product itself, packaging, containers, labels, and tags, and examples of indirect labeling include advertising and promotional activities by place or means such as notifications by apps in AI diagnosis, websites, stores, pamphlets, exhibitions, seminars such as media seminars, books, newspapers, magazines, television, radio, mail, email, and voice.

[0082] Examples of uses and functions that can be indicated for a food composition include promotion of ghrelin secretion, the ability to treat diseases or conditions that are improved by promotion of ghrelin secretion, such as promotion of growth hormone secretion, promotion of fat accumulation (suppression of weight loss), suppression of insulin secretion, suppression of brown adipose tissue function, promotion of gastrointestinal motility, enhancement of gastric acid secretion, enhancement of pancreatic exocrine secretion, vasodilation, increase in cardiac contractility, and protection of cardiomyocytes, as well as the ability to regulate, support, maintain, and use as a probiotic.

[0083] In one embodiment, the recommendation to take the composition is displayed personally. Such a display can be made using a document (whether written or electronic) addressed to the subject, the subject's tablet terminal, smartphone, personal computer, the subject's SNS, etc. Also, such a display can be made together with the display of the results of any test or analysis, such as a blood test or a blood metabolome test, for the subject.

[0084] In one aspect, a method for promoting ghrelin secretion in a subject is provided, comprising a step of having a subject ingest a composition containing a bacterium belonging to Lactobacillus delbrueckii based on the amount of ghrelin in the subject's blood (blood concentration of ghrelin). In another aspect, a method for promoting fat accumulation (suppressing weight loss), suppressing insulin secretion, suppressing brown adipose tissue function, promoting gastrointestinal motility, enhancing gastric acid secretion, enhancing pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility, or protecting cardiomyocytes in a subject is provided, comprising a step of having a subject ingest a composition containing a bacterium belonging to Lactobacillus delbrueckii based on the amount of ghrelin in the subject's blood (blood concentration of ghrelin). In yet another aspect, a method for suggesting to a subject to ingest a composition, a method for supporting a subject's diet, or a method for supporting a subject's health is provided, comprising a step of suggesting to a subject to ingest a composition containing a bacterium belonging to Lactobacillus delbrueckii based on the amount of ghrelin in the subject's blood (blood concentration of ghrelin). In these methods, the subject's blood is preferably collected in a fasting state. The blood may be separated plasma. In these methods, the amount of ghrelin may be either or both of acylated ghrelin (active form) and non-acylated ghrelin. The amount of ghrelin can be measured by quantifying ghrelin in blood, performing metabolomic analysis of blood, etc. The amount of ghrelin can be quantified by any method known to those skilled in the art, such as ELISA or HPLC. In these methods, the subject may be, for example, a person whose fasting blood ghrelin concentration is 5 pg / mL or less. These methods may, for example, further comprise the steps of: A step of collecting blood from a subject; a step of measuring the amount of ghrelin in the blood; a step of displaying the measurement results; a step of determining whether it is desirable or necessary to promote ghrelin secretion in the subject based on the measurement results; a step of displaying the determination results; and a step of displaying a product which is a composition containing bacteria belonging to Lactobacillus delbrueckii. EXAMPLES

[0085] [Example 1. Examination of the ghrelin secretion promoting effect of Lactobacillus delbrueckii ssp. bulgaricus species] MKN45 cells were cultured in antibiotic-free RPMI-1640 medium on a 24-well plate for 24 hours. The Lactobacillus delbrueckii ssp. bulgaricus strains shown in Table 1 below were subjected to activation culture and main culture for 20 hours in MRS medium.

[0086] The cultured cells were washed with PBS and then cultured at OD 660= 0.5, and live bacteria were added to the plate. After 4 and 6 hours of incubation, the cells were washed three times with PBS and the Maxwell RSC simplyRNA Cells Kit (Promega) reagents T / E solution 200 μL and Lysis Buffer 200 μL were added to recover the cells. RNA was then extracted and cDNA was made, and the expression level of the ghrelin gene was measured by qPCR (SYBR Green intercalation method). Normalization was performed with β-actin, and analysis was performed using the ΔΔCt method, a comparative quantitative method.

[0087] [Table 1]

[0088] The results are shown in Figure 1. Compared to the control group to which no live bacteria was added, ghrelin mRNA expression levels were significantly increased in the 2038 and OLL205013 groups 4 hours after addition, and were also significantly increased in the P230250, OLL1255, OLL1171, and OLL1247 groups 6 hours after addition.

[0089] [Example 2. Examination of the ghrelin secretion promoting effect of various bacteria belonging to Lactobacillus delbrueckii] The ghrelin secretion ability was examined in the same manner as in Example 1 using the strains of Lactobacillus delbrueckii, Lactobacillus delbrueckii ssp. delbrueckii, Lactobacillus delbrueckii ssp. indicus, Lactobacillus delbrueckii ssp. lactis, or Lactobacillus delbrueckii ssp. sunkii shown in Table 2 below.

[0090] [Table 2]

[0091] The results are shown in Figure 2. Compared to the control group to which no live bacteria was added, ghrelin mRNA expression levels were significantly increased in all groups 4 hours after addition, and significantly increased in all groups except the OLL204989 group 6 hours after addition.

[0092] As described above, since the strain belonging to L. delbrueckii promoted the expression of the ghrelin gene, it can be said that the bacteria belonging to L. delbrueckii can be used to promote the secretion of ghrelin. In addition, ghrelin is known to have actions such as promoting growth hormone secretion (Patent Documents 1 and 2), promoting fat accumulation (suppressing weight loss), suppressing insulin secretion, suppressing the function of brown adipose tissue, promoting gastrointestinal motility, enhancing gastric acid secretion, enhancing exocrine pancreatic secretion, vasodilation, increasing cardiac contractility, and protecting myocardial cells (Patent Document 3). Therefore, it can be said that the bacteria belonging to L. delbrueckii can be used for any one or more selected from the group consisting of promoting growth hormone secretion, promoting fat accumulation (suppressing weight loss), suppressing insulin secretion, suppressing the function of brown adipose tissue, promoting gastrointestinal motility, enhancing gastric acid secretion, enhancing exocrine pancreatic secretion, vasodilation, increasing cardiac contractility, and protecting myocardial cells. [Industrial Applicability]

[0093] The present invention provides a ghrelin secretion promoter derived from a highly safe food ingredient or a material whose safety has been confirmed, thereby supporting the maintenance and improvement of health and work efficiency in people who are not yet ill. Furthermore, the present invention can provide a food composition and a method for producing a food that are useful for promoting ghrelin secretion or that are useful for supporting the maintenance and improvement of health and work efficiency in people who are not yet ill.

Claims

1. A composition for promoting ghrelin secretion, comprising bacteria belonging to the genus Lactobacillus delbrueckii.

2. A composition comprising a bacterium belonging to Lactobacillus delbrueckii for one or more effects selected from the group consisting of promotion of growth hormone secretion, inhibition of brown adipose tissue function, promotion of gastrointestinal motility, enhancement of gastric acid secretion, enhancement of pancreatic exocrine secretion, vasodilation, increase in cardiac contractility, and protection of myocardial cells.

3. The composition of claim 2, wherein any one or more of the effects selected from the group consisting of promotion of growth hormone secretion, inhibition of brown adipose tissue function, promotion of gastrointestinal motility, increased gastric acid secretion, increased pancreatic exocrine secretion, vasodilation, increased cardiac contractility, and protection of cardiomyocytes are due to promotion of ghrelin secretion.

4. The composition according to any one of claims 1 to 3, wherein the bacterium belonging to Lactobacillus delbrueckii belongs to any one of Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus delbrueckii ssp. delbrueckii, Lactobacillus delbrueckii ssp. indicus, Lactobacillus delbrueckii ssp. lactis, and Lactobacillus delbrueckii ssp. sunkii.

5. The composition according to any one of claims 1 to 3, wherein the bacterium belonging to Lactobacillus delbrueckii has a 16S rRNA gene having 90% or more identity to a base sequence represented by any one of SEQ ID NOs: 1 to 5.

6. Bacteria belonging to the genus Lactobacillus delbrueckii ssp. bulgaricus were identified as Lactobacillus delbrueckii ssp. bulgaricus 2038, Lactobacillus delbrueckii ssp. bulgaricus OLL1255 (Accession Number: NITE BP-76), Lactobacillus delbrueckii ssp. bulgaricus OLL1171 (Accession Number: NITE BP-01569), Lactobacillus delbrueckii ssp. bulgaricus OLL1247 (Accession Number: NITE BP-01814), Lactobacillus delbrueckii ssp. bulgaricus OLL205013 (Accession Number: NITE BP-02411), and Lactobacillus delbrueckii OLL204989 (Accession Number: NITE BP-02412). BP-02874), Lactobacillus delbrueckii ssp. delbrueckii (JCM 1012T), Lactobacillus delbrueckii ssp. indicus (JCM 15610T), Lactobacillus delbrueckii ssp. lactis (JCM 1248T), and Lactobacillus delbrueckii ssp. sunkii (JCM 17838T). The composition according to any one of claims 1 to 3,

7. The composition according to any one of claims 1 to 3, wherein the bacterium belonging to Lactobacillus delbrueckii is a live cell.

8. The composition according to any one of claims 1 to 3, which is a fermented milk that does not contain bacteria belonging to Lactobacillus paragaceri.

9. The composition according to any one of claims 1 to 3, wherein the bacterium belonging to Lactobacillus delbrueckii does not have a DNA consisting of the sequence of SEQ ID NO:

6.

10. A pharmaceutical composition comprising a bacterium belonging to the group Lactobacillus delbrueckii.

11. The pharmaceutical composition of claim 10 which is a hormonal agent.

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