CN118900639A - Composition for controlling proliferation of Collinsella bacteria and its application - Google Patents

Abstract

A means for controlling the proliferation of Collinsella bacteria in the intestine is provided. The subject is to provide a new material capable of controlling the proliferation of Collinsella bacteria. A composition for controlling the proliferation of Collinsella bacteria in the intestinal flora is provided, which comprises any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides.

Description

Composition for controlling proliferation of Collinsella bacteria and its application

Technical Field

The present invention relates to a composition for controlling the growth of bacteria belonging to the genus Collinsella and its application.

Background Art

It is known that Collinsella bacteria are intestinal bacteria present in the human intestine. Among them, Collinsella aerofaciens (C. aerofaciens) is a common intestinal anti-PD-1 antibody responder in cancer immunotherapy (non-patent document 1), and has key genes related to the effectiveness of fecal transplantation therapy for recurrent Clostridium difficile infection (non-patent document 2). In addition, it was recently reported that a machine learning model (generalized linear model; GLM) was created to predict the COVID-19 mortality rate in various countries using intestinal flora, and the results of the analysis using GLM showed that the lower the ratio of Collinsella in the intestinal flora, the higher the COVID-19 mortality rate (non-patent document 3).

Regarding Collinsella bacteria, whole fiber and inulin are known to be materials that can make Collinsella bacteria proliferate in the intestine (Non-Patent Document 4). In addition, Patent Document 1 describes a bile acid adsorption method, characterized in that intestinal bacteria having the property of taking bile acid into the bacteria in the presence of a sugar source are used, and Collinsella bacteria are recorded as one of the intestinal bacteria used. In addition, Patent Document 2 describes a composition for use in a method for treating intrahepatic cholestasis of a subject or reducing the risk of intrahepatic cholestasis, the composition comprising at least one bacterial strain having bile acid deconjugation activity and/or 7-diastereomerization activity, and Collinsella aerofaciens ATCC25986 is recorded as one of the bacterial strains contained in the composition. In addition, Patent Document 3 describes a method for producing L-ornithine and equol, which comprises: (1) a step of culturing anaerobic microorganisms in a culture medium containing isoflavones and dextrin; and (2) a step of recovering L-ornithine and equol obtained in step (1), wherein one of the anaerobic microorganisms is a bacterium of the genus Collinsella. Patent Document 4 describes a composition that can effectively treat and prevent inflammation, particularly ulcerative colitis, characterized in that it contains Megamonas funiformis and/or its metabolites and contains Anaerofustis stercorihominis and/or its metabolites, and the composition further contains Collinsella shenzhenensis and/or its metabolites, and preferably the above-mentioned Collinsella shenzhenensis is Collinsella shenzhenensis TF06-26 with a deposit number of GDMCC60090. Patent document 5 describes a human intestinal flora model for evaluating the effect of promoting the increase of short-chain fatty acids and/or the increase of diversity in the human intestine of a target food composition. The human intestinal flora model is composed of an evaluation bacterial culture containing at least 9 bacterial species, which does not contain bacteria belonging to the Proteobacteria or Lactobacillus genus and contains bacteria belonging to the Bifidobacterium genus. Collinsella bacteria are recorded as one of the bacteria that can be included in the model.

On the other hand, human milk oligosaccharides (HMO) are the third most abundant component in breast milk solids after lactose and lipids. It is known that if 2'-fucosyllactose, 3-fucosyllactose, and lactose-difucotetraose, which are human milk oligosaccharides, are provided, bifidobacteria will proliferate (Non-Patent Document 5).

Prior art literature

Patent Literature

Patent Document 1: Japanese Patent Application Publication No. 2006-314219

Patent Document 2: WO2017/102816 (Japanese Patent Publication No. 2018-537484)

Patent Document 3: Japanese Patent Application Publication No. 2014-54234

Patent Document 4: WO2019/227418 (Japanese Patent Publication No. 2021-524751)

Patent document 5: WO2021/206106

Non-patent literature

Non-patent literature 1: Matson V, Fessler J, Bao R, et al. The common microbiomes associated with anti-PD-1 efficacy in metastatic melanoma patients. Science. 2018; 359(6371): 104-108. doi: 10.1126/science.aao3290

Non-patent literature 2: Mullish BH, McDonald JAK, Pechlivanis A, et al. Microbialbile salt hydrolases mediate the efficacy of faecal microbiota transplant in the treatment of recurrent Clostridioides difficile infection. Gut.2019;68(10):1791-1800.doi: 10.1136/gutjnl-2018-317842

Non-patent document 3: Hirayama1 M, Nishiwaki H, et al.Intestinal Collinsella maymitigate infection and exacerbation of COVID-19 by producingursodeoxycholate.PLoS One.2021Nov 23;16(11):e0260451.doi:10.1371/journal.pone.0260451.eCollection 2021.

Non-patent document 4: Carlson JL, Erickson JM, Hess JM, Gould TJ, SlavinJL. Prebiotic Dietary Fiber and Gut Health: Comparing the in VitroFermentations of Beta-Glucan, Inulin and Xylooligosaccharide. Nutrients. 2017; 9(12): 1361. Published 2017Dec 15.doi:10.3390/nu9121361

Non-patent document 5: Yu ZT, Chen C, Newburg DS. Utilization of major fucosylated and sialylated human milk oligosaccharides by isolated human gutmicrobes. Glycobiology. 2013; 23(11):1281-1292.doi:10.1093/glycob/cwt065

Summary of the invention

Problem that the invention aims to solve

Collinsella bacteria are considered to be one of the useful intestinal bacteria. New materials that can promote the growth of Collinsella bacteria in the intestine are desired.

Solutions for solving problems

The present invention provides the following:

[1] A composition for suppressing the growth of Collinsella bacteria in intestinal flora, the composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides.

[2] The composition according to item 1, comprising any one selected from the group consisting of fucosylated human milk oligosaccharides, sialylated human milk oligosaccharides, human milk oligosaccharides containing lactose as a constituent sugar, human milk oligosaccharides containing sialic acid as a constituent sugar, lactose and sialic acid.

[3] The composition according to 1 or 2, comprising any one selected from the group consisting of 2'-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, lactose, and N-acetylneuraminic acid.

[4] The composition according to any one of 1 to 3, which is used as a prebiotic or synbiotic.

[5] A composition for treating a disease or condition that is improved by controlling the growth of Collinsella bacteria in the intestine, comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides.

[6] The composition according to 5, wherein the disease or condition improved by controlling the proliferation of Collinsella bacteria in the intestine is a respiratory infection.

[7] A composition for treating a respiratory infection, comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides.

[8] The composition according to 6 or 7, wherein the respiratory infection is a coronavirus infection.

[9] The composition according to 6 or 7, wherein the respiratory infection is novel coronavirus infection (COVID-19).

[10] The composition according to any one of 1 to 5, wherein the proliferation control is proliferation promotion.

[11] A composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides, for use in a method for controlling the proliferation of Collinsella bacteria in the intestinal flora. Use of any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides in the manufacture of a composition for controlling the proliferation of Collinsella bacteria in the intestinal flora. A method or non-therapeutic method for controlling the proliferation of Collinsella bacteria in the intestinal flora, comprising the following steps: administering a composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides to a subject. Use or non-therapeutic use of a composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides for controlling the proliferation of Collinsella bacteria in the intestinal flora.

[12] The composition according to item 11, comprising any one selected from the group consisting of fucosylated human milk oligosaccharides, sialylated human milk oligosaccharides, human milk oligosaccharides with lactose as a constituent sugar, human milk oligosaccharides with sialic acid as a constituent sugar, lactose and sialic acid; use of any one selected from the group consisting of fucosylated human milk oligosaccharides, sialylated human milk oligosaccharides, human milk oligosaccharides with lactose as a constituent sugar, human milk oligosaccharides with sialic acid as a constituent sugar, lactose and sialic acid in the manufacture according to item 11; the method or non-therapeutic method according to item 11, comprising the following step: administering the composition comprising any one selected from the group consisting of fucosylated human milk oligosaccharides, sialylated human milk oligosaccharides, human milk oligosaccharides with lactose as a constituent sugar, human milk oligosaccharides with sialic acid as a constituent sugar, lactose and sialic acid to a subject; or the use or non-therapeutic use of such a composition according to item 11.

[13] The composition according to 7 or 8, comprising any one selected from the group consisting of 2'-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, lactose and N-acetylneuraminic acid; or, use of any one selected from the group consisting of 2'-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, lactose and N-acetylneuraminic acid in the manufacture according to 11 or 12; the method or non-therapeutic method according to 11 or 12, comprising the following step: administering a composition comprising any one selected from the group consisting of 2'-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, lactose and N-acetylneuraminic acid to a subject; or, the use or non-therapeutic use of such a composition according to 11 or 12.

[14] The composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use according to any one of 11 to 13, wherein the composition is used as a prebiotic or synbiotic.

[15] A composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides, for use in a method for treating a disease or condition that is improved by controlling the growth of Collinsella bacteria in the intestine. Use of any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides in the manufacture of a composition for treating a disease or condition that is improved by controlling the growth of Collinsella bacteria in the intestine. A method or non-therapeutic method for treating a disease or condition that is improved by controlling the growth of Collinsella bacteria in the intestine, comprising the following steps: administering a composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides to a subject. Use or non-therapeutic use of a composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides for treating a disease or condition that is improved by controlling the growth of Collinsella bacteria in the intestine.

[16] The composition, the use, method or non-therapeutic method for the manufacture of the composition, or the use or non-therapeutic use according to item 15, wherein the disease or condition improved by controlling the proliferation of Collinsella bacteria in the intestine is a respiratory infection.

[17] A composition for treating a respiratory infection, comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides. A composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides, for use in a method for treating a respiratory infection. A composition for treating a respiratory infection, comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides. Use in the manufacture of a composition for treating a respiratory infection. A method or non-therapeutic method for treating a respiratory infection, comprising the step of administering a composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides to a subject. Use or non-therapeutic use of a composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides for treating a respiratory infection.

[18] The composition, the use, method or non-therapeutic method for the manufacture of the composition, or the use or non-therapeutic use according to item 16 or 17, wherein the respiratory infection is a coronavirus infection.

[19] The composition, the use, method or non-therapeutic method in the manufacture of the composition, or the use or non-therapeutic use according to any one of 16 to 18, wherein the respiratory infection is the novel coronavirus infection (COVID-19).

[20] The composition, the use, method or non-therapeutic method for the manufacture of the composition, or the use or non-therapeutic use according to any one of 11 to 16, wherein the proliferation control is proliferation promotion.

Effects of the Invention

The present invention can control the flora in the intestine, and in particular, can control the increase of Collinsella bacteria in the flora in the intestine.

By controlling the increase of Collinsella bacteria in the intestinal flora, it is expected that various diseases or conditions that can be improved by this, such as respiratory infections such as COVID-19, can be treated.

By adding the active ingredient of the present invention, functional foods in various forms can be provided.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows the changes in the occupancy of Collinsella caused by various sugars in the human feces culture system. Based on paired t-test.

DETAILED DESCRIPTION

The present invention is described in detail below.

The present invention relates to a composition for suppressing the growth of Collinsella bacteria in intestinal flora, comprising as an active ingredient any one selected from the group consisting of human milk oligosaccharides (also referred to as HMO) and constituent sugars of human milk oligosaccharides.

[Active ingredients]

The composition of the present invention comprises any one selected from the group consisting of human milk oligosaccharides (sometimes also referred to as HMO) and constituent sugars of human milk oligosaccharides as an active ingredient. Human milk oligosaccharides are oligosaccharides contained in human milk and are the third most abundant solid component after lactose and lipids. Human milk oligosaccharides, with a few exceptions, have the following chemical structure: a lactose unit is present at the reducing end, and fucose, sialic acid, galactose, etc. are added thereto.

It should be noted that, in the present invention, the abbreviations and structures of milk oligosaccharides follow Nutr Rev. 2017 Nov 1; 75(11): 920-933. doi: 10.1093/nutrit/nux044. and Nutrients 2021, 13(8), 2737; https://doi.org/10.3390/nu13082737. If there is a disagreement between these documents, the latter shall be followed.

The human milk oligosaccharides and the constituent sugars of human milk oligosaccharides used in the composition are not particularly limited as long as they have the intended effect. In addition, the human milk oligosaccharides and the constituent sugars of human milk oligosaccharides used in the composition may be one type or a combination of two or more types.

Examples of the human milk oligosaccharide contained in the composition include the following neutral oligosaccharides and acidic oligosaccharides.

Neutral oligosaccharides: 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), difucosyllactose (Lacto-difucotetraose) (DFL, LDFT), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), lactose-N-fucopentaose I (LNFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-difucohexaose I (LNDFHI), lactose-N-difucohexaose II (LNDFH II), lactose-N-hexaose (LNH), para-lactose-N-hexaose (pLNH), lactose-N-neohexose (LNnH), para-lactose-N-neohexose (pLNnH), fucosyllactose-N-hexaose I (F-LNH I), fucosyl-para-lactose-N-hexaose I (F-para-LNH I), fucosyllactose-N-hexaose II (F-LNH II), fucosyl-lactose-N-hexaose III (F-LNH III), difucosyl-lactose-N-hexaose I (DF-LNH I), difucosyl-lactose-N-hexaose II (DF-LNH II), difucosyl-lactose-N-hexaose III (DF-LNH III), difucosyl-p-lactose-N-neohexose (DF-pLNnH), difucosyl-p-lactose-N-hexaose (DF-para-LNH), difucosyl-p-lactose-N-neohexose (DF-para-LNnH), trifucosyl-lactose-N-hexaose (TF-LNH).

Acidic oligosaccharides: 3'-sialyllactose (3'-SL), 6'-sialyllactose (6'-SL), 6'-sialyllactose-N-acetyllactosamine (6'-SLN), LS-tetrasaccharide a (sialyllactose-N-tetrasaccharide a) (LST a), LS-tetrasaccharide b (sialyllactose-N-tetrasaccharide b) (LST b), LS-tetrasaccharide c (sialyllactose-N-tetrasaccharide c) (LST c), disialyllactose-N-tetrasaccharide (DS-LNT), fucosyl-sialyllactose-N-tetrasaccharide a (F-LST a), fucosyl-sialyllactose-N-tetrasaccharide b (F-LST b), fucosyl-sialyllactose-N-hexasaccharide (FS-LNH), fucosyl-sialyllactose-N-neohexacharide (FS-LNNH I), fucosyl-disialyl lactose-N-hexaose (FDS-LNH), disialyl lactose-N-hexaose (DS-LNH).

Preferred examples of the human milk oligosaccharide contained in the composition include the following neutral oligosaccharides and acidic oligosaccharides.

Neutral oligosaccharides: 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), difucosyllactose (lactose-difucotetraose) (DFL, LDFT), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), lactose-N-fucopentaose I (LNFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-difucohexaose I (LNDFH I), lactose-N-difucohexaose II (LNDFH II), lactose-N-hexaose (LNH), lactose-N-neohexose (LNnH), para-lactose-N-neohexose (pLNnH), fucosyllactose-N-hexaose I (F-LNH I), fucosyl-para-lactose-N-hexaose I (F-para-LNHI), fucosyllactose-N-hexaose II (F-LNH II), fucosyllactose-N-hexaose III (F-LNH III), difucosyllactose-N-hexaose I (DF-LNH I), difucosyllactose-N-hexaose II (DF-LNH II), difucosyllactose-N-hexaose III (DF-LNH III), difucosyllactose-para-lactose-N-neohexose (DF-pLNnH), trifucosyllactose-N-hexaose (TF-LNH).

Acidic oligosaccharides: 3’-sialyllactose (3’-SL), 6’-sialyllactose (6’-SL), LS-tetrasaccharide a (sialyllactose-N-tetrasaccharide a) (LST a), LS-tetrasaccharide b (sialyllactose-N-tetrasaccharide b) (LST b), LS-tetrasaccharide c (sialyllactose-N-tetrasaccharide c) (LST c), disialyllactose-N-tetrasaccharide (DS-LNT), fucosyl-sialyllactose-N-neohexaose (FS-LNnH I), disialyllactose-N-hexaose (DS-LNH).

More preferable examples of the human milk oligosaccharide contained in the composition include the following neutral oligosaccharides and acidic oligosaccharides.

Neutral oligosaccharides: 2'-fucosyllactose (2'-FL), difucosyllactose (lactose-difucotetraose) (DFL, LDFT), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT)

Acidic oligosaccharides: 3'-sialyllactose (3'-SL), 6'-sialyllactose (6'-SL)

In the present invention, when referring to the constituent sugars of human milk oligosaccharides, it means: sugars constituting human milk oligosaccharides (sugars produced by the decomposition of human milk oligosaccharides) excluding glucose and galactose. Examples of the constituent sugars of human milk oligosaccharides contained in the composition include lactose, fucose, sialic acid, and N-acetylglucosamine. Sialic acid is a general term for modified forms of neuraminic acid. As an example of sialic acid, N-acetylneuraminic acid (Neu5Ac) formed by modifying neuraminic acid with an acetyl group can be listed.

According to the research of the inventors, it is confirmed that lactose, Neu5Ac, 2'-FL, 3'-SL, and 6'-SL have excellent Collinsella bacterial proliferation promoting effects. Here, lactose may be generated in the human digestive tract after ingesting human milk oligosaccharides. In addition, after ingesting human milk oligosaccharides containing human milk oligosaccharides containing Neu5Ac as a constituent sugar, Neu5Ac may be generated in the human digestive tract (refer to non-patent documents 6 to 8 described later). Furthermore, the target effect was confirmed for each of 2'-FL, 3'-SL, and 6'-SL, so if it is fucosylated human milk oligosaccharides and sialylated human milk oligosaccharides, the target effect can be fully obtained.

From this viewpoint, the human milk oligosaccharides used in the composition and the preferred examples of the constituent sugars of the human milk oligosaccharides are any selected from the group consisting of fucosylated human milk oligosaccharides, sialylated human milk oligosaccharides, human milk oligosaccharides with lactose as a constituent sugar, human milk oligosaccharides with sialic acid as a constituent sugar, lactose and sialic acid. Human milk oligosaccharides with sialic acid as a constituent sugar include sialylated human milk oligosaccharides (e.g., 3'-SL, 6'-SL) and sialic acid-containing human milk oligosaccharides (e.g., LST a, DS-LNT).

Specific examples of human milk oligosaccharides and constituent sugars of human milk oligosaccharides used in the composition are any selected from the group consisting of 2'-FL, 3'-SL, 6'-SL, lactose, and Neu5Ac.

[use]

The composition of the present invention can be used to control the proliferation of Collinsella bacteria in the intestinal flora. Proliferation control includes proliferation promotion and proliferation inhibition. The preferred composition is used for proliferation promotion.

Examples of bacteria contained in the intestinal flora include the following:

Bacteria of the genus Faecalibacterium, for example, Faecalibacterium prausnitzii;

Bacteria of the genus Akkermansia, for example Akkermansia muciniphila;

Bacteria of the genus Blautia, for example Blautia coccoides, Blautia intestinalis, Blautia faecicola;

Bacteria of the genus Subdoligranulum, for example Subdoligranulumvariabile;

Bacteria of the genus Bilophila, for example, Bilophila wadsworthia;

Bacteria of the genus Collinsella, for example, Collinsella aerofaciens and Collinsella intestinalis;

Bacteria of the genus Holdemania, for example, Holdemania filiformis;

Bacteria of the genus Parasutterella, for example, Parasutterella excrementihominis, Parasutterella secunda;

Oscillibacter bacteria, such as Oscillibacter valericigenes;

Bacteria of the genus Eggerthella, for example Eggerthella lenta;

Bacteria of the genus Sutterella, for example, Sutterella parvirubra, Sutterella wadsworthensis;

Bacteria of the genus Bifidobacterium (bifidobacteria), for example, Bifidobacterium adolescentis (B. adolescentis), Bifidobacterium bifidum (B. bifidum), Bifidobacterium breve (B. breve), Bifidobacterium catenulatum (B. catenulatum), Bifidobacterium infantis (B. infantis), Bifidobacterium lactis (B. lactis), Bifidobacterium longum (B. longum), Bifidobacterium pseudocatenulatum (B. pseudocatenulatum).

In the present invention, when referring to Collinsella bacteria, it refers to bacteria identified as Collinsella by molecular phylogenetic analysis based on 16S rRNA gene. The standard for judging the genus by molecular phylogenetic analysis based on 16S rRNA gene is well known to those skilled in the art (Stackebrandt E, Ebers J. Taxonomic parameters revisited: tarnished gold standards. Microbiol Today 2006; 33: 152-155.).

In the present invention, controlling the growth of specific bacteria in the bacterial flora means controlling the ratio (occupancy rate) of the specific bacteria in the bacterial flora.

Whether a certain component controls the proliferation of specific bacteria in the intestinal flora can be evaluated as follows. Add feces provided by a healthy person to a suitable culture medium, culture for a certain period of time as needed, add the component to be evaluated, culture under suitable conditions (for example, culture for 48 hours at 37°C and anaerobic conditions similar to the conditions in the intestine), and measure the occupancy rate of specific bacteria in the flora in the culture. Then, compare the measurement results with the measurement results of a culture cultured under the same conditions that differs only in that a control (such as sterilized water) is added instead of the component to be evaluated. When the occupancy rate is higher than the control, it can be judged that the proliferation is promoted, and when the occupancy rate is lower than the control, it can be judged that the proliferation is inhibited.

The occupancy rate of specific bacteria contained in the flora can be obtained by a known method. One of the preferred methods is to perform 16S metagenomic analysis (sequence analysis of 16S rRNA gene amplicon) on DNA extracted from the culture. In the case of implementing 16S metagenomic analysis, DNA extraction can be performed using a commercially available kit. The genomic region analyzed is not particularly limited as long as the bacteria can be determined, but the V3-V4 region of the 16S rRNA gene can be used. Methods such as primers, amplification conditions, and purification of amplicon for bacterial analysis can also use methods well known to those skilled in the art. The interpretation of the sequence is preferably performed with a next-generation sequencer with higher performance. The data analysis obtained can utilize a new generation of microbiome bioinformatics platforms such as QIIME 2 ^TM . Regarding 16S metagenomic analysis, those skilled in the art may refer to Sanschagrin S, Yergeau E. Next-generation sequencing of 16S ribosomal RNA geneamplicons. J Vis Exp. 2014; (90): 51709. Published 2014 Aug 29. doi: 10.3791/51709 and other information.

The Collinsella bacteria involved in the present invention are intestinal bacteria present in the human intestine, among which Collinsella aerogenes is a common intestinal anti-PD-1 antibody responder in cancer immunotherapy (Non-Patent Document 1) and has a key gene related to the effectiveness of fecal transplantation therapy for recurrent Clostridium difficile infection (Non-Patent Document 2), which is well known. Therefore, the composition can be expected to be used for the treatment of such diseases or conditions.

In addition, the composition can be used for treating diseases or conditions that can be improved by controlling the growth of Collinsella bacteria in the intestine. Such diseases or conditions may be various diseases or conditions.

Recently, it has been reported that the lower the ratio of Collinsella in the intestinal flora, the higher the mortality rate of COVID-19 (non-patent document 3). And it is known that Collinsella has genes related to the production of ursodeoxycholic acid as a secondary bile acid (non-patent document 9), and ursodeoxycholic acid is known to prevent the binding of the first receptor angiotensin-converting enzyme 2 (ACE2) when infected with SARS-CoV-2 (non-patent documents 10, 11). It is also known that ursodeoxycholic acid inhibits inflammation-induced cytokines (non-patent documents 12, 13), has antioxidant/anti-apoptotic effects (non-patent documents 14, 15), and improves alveolar fluid clearance in the case of acute respiratory syndrome (ARDS) (non-patent document 16). Therefore, a composition that controls the proliferation of Collinsella in the intestine can be used for the treatment of respiratory infections such as COVID-19 infection.

Non-patent literature 9. Liu L, Aigner A, Schmid RD. Identification, cloning, heterologous expression, and characterization of a NADPH-dependent7β-hydroxysteroid dehydrogenase from Collinsella aerofaciens. Appl MicrobiolBiotechnol. 2011; 90(1):127-135.doi :10.1007/s00253-010-3052-y

Non-patent literature 10. Poochi SP, Easwaran M, Balasubramanian B, Anbuselvam M, Meyyazhagan A, Park S, et al. Employing bioactive compounds derived from Ipomoeaobscura(L.) to evaluate potential inhibitor for SARSCoV-2main protease andACE2 protein.Food Front .2020.Epub 2020/08/25.https://doi.org/10.1002/fft2.29PMID:32838301;PubMed Central PMCID:PMC7361879.

Non-patent literature 11. Carino A, Moraca F, Fiorillo B, Marchiano S, Sepe V, Biagioli M, et al. Hijacking SARS-CoV-2/ACE2 Receptor Interaction by Natural and Semi-synthetic Steroidal Agents Acting on Functional Pockets on the ReceptorBinding Domain.Front Chem.2020;8:572885.Epub 2020/11/17.doi:10.3389/fchem.2020.572885;PubMed Central PMCID:PMC7645072.

Non-patent literature 12.Ko WK, Lee SH, Kim SJ, Jo MJ, Kumar H, Han IB, et al. Anti-inflammatory effects of ursodeoxycholic acid by lipopolysaccharide-stimulated inflammatory responses in RAW 264.7macrophages.PLoS One.2017; 12( 6):e0180673.Epub 2017/07/01.doi:10.1371/journal.pone.0180673; PubMed Central PMCID: PMC5493427.

Non-patent literature 13. Ko WK, Kim SJ, Jo MJ, Choi H, Lee D, Kwon IK, etal. Ursodeoxycholic Acid Inhibits Inflammatory Responses and PromotesFunctional Recovery After Spinal Cord Injury in Rats. Mol Neurobiol. 2019; 56(1): 267–77.Epub 2018/04/25.doi:10.1007/s12035-018-0994-z.

Non-patent literature 14. Lapenna D, Ciofani G, Festi D, Neri M, Pierdomenico SD, Giamberardino MA, et al. Antioxidant properties of ursodeoxycholic acid. BiochemPharmacol. 2002; 64(11): 1661–7. Epub 2002/11/ 14.doi:10.1016/s0006-2952(02)01391-6.

Non-patent literature 15. Kim YJ, Jeong SH, Kim EK, Kim EJ, Cho JH. Ursodeoxycholicacid suppresses epithelial-mesenchymal transition and cancer stem cellformation by reducing the levels of peroxiredoxin II and reactive oxygen species in pancreatic cancer cells. Oncol Rep. 2017; 38(6):3632–8.Epub2017/11/14.doi:10.3892/or.2017.6045.

Non-patent literature 16. Niu F, Xu ):20057–65.Epub 2019/04/12.doi:10.1002/jcp.28602.

With regard to the present invention, when referring to respiratory infections, it includes infections caused by any virus selected from the group consisting of coronavirus, influenza virus, rhinovirus, RS virus, parainfluenza virus and adenovirus. In one embodiment, the composition can be used for coronavirus infections in respiratory infections. Coronavirus infections include infections caused by any virus selected from the group consisting of HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV) and new coronavirus (SARS-CoV-2). New coronavirus infections are also referred to as COVID-19. In one preferred embodiment, the composition is used for the treatment of new coronavirus infections (COVID-19).

With regard to the present invention, when referring to the treatment of a disease or condition, it refers to the reduction of the risk of disease, the delay of disease onset, prevention, treatment, cessation of disease progression, and delay. Treatment includes radical treatment (treatment to eliminate the cause of the disease) and symptomatic treatment (treatment to improve symptoms). The behavior used to improve or treat includes medical behavior performed by doctors or nurses, midwives, etc. who receive instructions from doctors, and non-therapeutic behavior performed by personnel other than doctors, such as pharmacists, nutritionists (including management nutritionists, sports nutritionists), health workers, midwives, nurses, clinical examination technicians, sports trainers, drug manufacturers, drug sellers, food manufacturers, food sellers, etc. Furthermore, preventing or reducing the risk of disease includes recommending the intake of specific foods and nutritional guidance (including nutritional guidance required for the recuperation of the wounded and sick, and nutritional guidance for maintaining and improving health).

[Composition]

(Food composition, etc.)

The composition of the present invention can be made into a food composition or a pharmaceutical composition. Unless otherwise specified, food and medicine include not only food and medicine for human use, but also food and medicine for animals other than humans. Unless otherwise specified, food includes general food, functional food, nutritional composition, and also includes therapeutic food (food for therapeutic purposes. Food cooked based on a diet recipe prescribed by a doctor and a menu prepared by a nutritionist, etc.), dietetic food, formula food, nursing food, and therapeutic auxiliary food. Unless otherwise specified, food includes not only solid food, but also liquid food, such as beverages, health drinks, liquid food and soup. Functional food refers to food that can give a specified functionality to an organism, such as specific health food (including special protection [specific health food] with additional conditions), functional labeled food, health functional food including nutritional functional food, special purpose food, nutritional supplement food, health supplement food, nutritional supplement (such as tablets, coated tablets, sugar-coated tablets, capsules, liquid preparations and other various dosage forms of food), beauty food (such as weight loss food) and other all health foods. In addition, the "functional food" in the present invention includes health foods with health claims based on applicable food standards of the Codex (FAO/WHO Joint Food Standards Commission).

(Target)

The composition of the present invention is suitable for administration to the following subjects: subjects who wish or need to control the proliferation of Collins bacteria in the intestine; and subjects in a disease or state that can be improved by controlling the proliferation of Collins bacteria in the intestine; those at risk of contracting pathogenic viruses of respiratory infections, those at risk of contracting coronaviruses such as SARS-CoV-2, patients with respiratory infections, and patients with coronavirus infections such as COVID-19. Subjects who wish or need to control the proliferation of Collins bacteria in the intestine include those with a small number of Collins bacteria in the intestine. It should be noted that, with respect to the present invention, administration is used in the sense of administering a drug to a subject, and is also used in the sense of causing a subject to ingest food other than a drug.

The age of the subjects is not particularly limited, and the subjects may be, for example, newborns (within 28 days of birth); infants (under 1 year old); toddlers (1 to 6 years old); children (over 7 years old and under 15 years old); adults (over 15 years old); and people over 65 years old.

(Given by, etc.)

The composition of the present invention may be administered orally, parenterally, for example, via a tube (gastrostomy, enterostomy), or nasally, but is preferably administered orally.

The composition can be administered to the subject repeatedly or continuously for a long period of time. The duration is not particularly limited, but in order to fully observe the effect, it can be administered continuously for a relatively long period of time, such as more than 3 days, more than 1 week, more than 2 weeks, more than 1 month, more than 3 months, more than 6 months, or more than 1 year.

The composition can be administered daily, in advance, or when needed, etc. The composition can be administered as a meal, before, after, or between meals, or when the disease or condition to be improved by the composition occurs.

(Dosage, content)

The dosage of the composition of the present invention may be any dosage that can produce the desired effect. The dosage may be appropriately set in consideration of various factors such as the age, weight, and symptoms of the subject.

The daily dose of the composition can be such that the amount of the active ingredient is more than 0.1 mg, preferably more than 0.3 mg, more preferably more than 0.6 mg, and more preferably more than 1 mg. The upper limit of the active ingredient of the daily dose can be set to less than 10 g, less than 5 g, less than 1 g, less than 100 mg, less than 60 mg, less than 30 mg, or less than 15 mg in any case where the lower limit is any. It should be noted that when a plurality of active ingredients are included in the composition, the amount of the active ingredient refers to the total amount of the active ingredient contained.

Administration may be once a day or multiple times a day, for example 2 to 10 times. The dose of the active ingredient each time may be, for example, 0.01 mg or more, preferably 0.03 mg or more, more preferably 0.06 mg or more, and further preferably 0.1 mg or more. The upper limit of the active ingredient each time may be 3 g or less, 1 g or less, 100 mg or less, 33 mg or less, 20 mg or less, 10 mg or less, or 5 mg or less, when the lower limit is any.

The content of the active ingredient in the composition can be appropriately set according to the form of the composition. For example, relative to the solid content of the composition, the content of the active ingredient can be set to 0.1% or more, preferably 0.3% or more, more preferably 0.6% or more, and further preferably 1% or more. The upper limit value of the active ingredient relative to the solid content can be set to 50% or less, 30% or less, 20% or less, 10% or less, or 5% or less, when the lower limit value is arbitrary. It should be noted that, with respect to the present invention, unless otherwise specified, % refers to mass %.

When the composition is in liquid form, the content of the active ingredient can be, for example, 0.01% or more, preferably 0.03% or more, more preferably 0.06% or more, and further preferably 0.1% or more. When the composition is in liquid form, the upper limit of the content of the active ingredient can be 5% or less, 3% or less, 2% or less, 1% or less, or 0.5% or less, with the lower limit being any.

(Other ingredients, additives)

In the present invention, the composition may contain other effective ingredients and nutritional ingredients that are permitted as food or medicine. Examples of such ingredients are lipids (e.g., milk fat, vegetable oils and fats, oils and fats containing medium-chain fatty acids), proteins (e.g., milk protein, milk protein concentrate (MPC), whey protein concentrate (WPC), whey protein isolate (WPI), α-lactalbumin (α-La), β-lactoglobulin (β-Lg), heat-denatured whey protein, and enzyme-treated whey protein), amino acids (e.g., lysine, arginine, glycine, alanine, glutamic acid, leucine, isoleucine, valine), and oligosaccharides other than human milk. Sugars or sugars other than the constituent sugars of human milk oligosaccharides (glucose, sucrose, fructose, maltose, trehalose, erythritol, maltitol, palatinose, xylitol, dextrin), electrolytes (such as sodium, potassium, calcium, magnesium), vitamins (such as vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, biotin, folic acid, pantothenic acid and niacin), minerals (such as copper, zinc, iron, cobalt, manganese), antibiotics, dietary fiber, etc.

In the present invention, the composition may contain prebiotics other than human milk oligosaccharides. Prebiotics can be said to be a kind of indigestible food ingredient that has a beneficial effect on the host and improves the health of the host by selectively changing the proliferation and activity of specific bacteria in the large intestine. As prebiotics other than human milk oligosaccharides in the composition, one or more prebiotics can be used.

Prebiotics other than human milk oligosaccharides are not particularly limited as long as they do not interfere with the effects of the active ingredients contained in the composition. Examples of prebiotics other than human milk oligosaccharides include galacto-oligosaccharides, fructo-oligosaccharides, xylo-oligosaccharides, isomaltooligosaccharides, raffinose, lactulose, lactofructooligosaccharides, soybean oligosaccharides, coffee oligosaccharides, dietary fiber, and gluconic acid.

In addition, the composition may also contain additives that are permitted as food or medicine. Examples of such additives include inactive carriers (solid, liquid carriers), excipients, surfactants, binders, disintegrants, lubricants, solubilizers, suspending agents, coating agents, colorants, preservatives, buffers, pH regulators, emulsifiers, stabilizers, sweeteners, antioxidants, flavors, acidulants, and natural products. More specifically, water, other aqueous solvents, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinyl pyrrolidone, 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, vaseline, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, sucralose, stevioside, aspartame, acesulfame potassium, citric acid, lactic acid, malic acid, tartaric acid, phosphoric acid, acetic acid, fruit juice, vegetable juice, etc.

(Dosage form/form)

The food composition of the present invention can be made into any form such as solid, liquid, mixture, suspension, powder, granule, paste, jelly, gel, capsule, etc. In addition, the food composition of the present invention can be made into any form such as dairy products, nutritional supplements, snacks, beverages, health beverages, flavorings, processed foods, side dishes, soups, etc. More specifically, the composition of the present invention can be made into liquid food, semi-liquid food, jelly, gel, powder, formula milk powder, formula liquid milk, milk powder/liquid milk for pregnant women/lactating women, fermented milk, stick, mousse, chocolate, biscuit, ice cream, fermented milk, lactobacillus beverage, milk beverage, milk beverage, refreshing beverage, sheet, cheese, bread, biscuit, cracker, pizza crust, patient food, nutritious food, frozen food, processed food, etc., and can also be made into forms such as granules, powders, pastes, concentrates, etc. for mixing in beverages and foods. Granules and powders can be made into cubes or sticks (small packages for single doses). In relation to the present invention, formula milk powder refers to a powdered product made by processing fresh milk, milk, special milk or fresh buffalo milk or foods made with them as raw materials or as the main raw materials, and adding nutrients required by infants and young children, as defined in Japan's "Ministerial Order on the Standards of Composition of Milk and Dairy Products (hereinafter referred to as "Ministerial Order on Milk, etc."). In relation to the present invention, formula liquid milk refers to a liquid product made by processing fresh milk, milk, special milk or fresh buffalo milk or foods made with them as raw materials or as the main raw materials, and adding nutrients required by infants and young children, as defined in the Provincial Order on Milk, etc.

Formula milk powder or formula liquid milk (sometimes they are collectively referred to as formula milk powder. In addition, in the description of formula milk powder, formula milk powder is sometimes used as an example, but this description is also applicable to formula liquid milk) can be for infants, older infants (for weaning and complementary food supplementation), low birth weight infants, children, adults, pregnant women, breastfeeding women, the elderly, allergic people, lactose intolerant people, and people with congenital metabolic disorders. Preferred examples of formula milk powder are milk powder for infants, older infants, low birth weight infants, and children.

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

(other)

In the manufacture of the composition of the present invention, the stage of compounding the active ingredient can be appropriately selected. As long as the characteristics of the active ingredient are not significantly damaged, the stage of compounding is not particularly limited. For example, the active ingredient can be mixed with the raw materials and compounded. Alternatively, the composition containing the active ingredient can be manufactured by adding the active ingredient in the final stage of the manufacture.

The composition of the present invention may be labeled with the intended use (application) and may also be labeled with content that recommends administration to a specific subject.

The composition of the present invention may be labeled as follows: the composition or active ingredient can control the proliferation of Collins bacteria in the intestine, can promote the proliferation of Collins bacteria in the intestine, can treat diseases or conditions that can be improved by controlling the proliferation of Collins bacteria in the intestine (preferably promoting proliferation), can be used as a prebiotic, can be used as a synbiotic (a combination of probiotics and prebiotics), or can treat related diseases, or can treat respiratory infections such as coronavirus infections. Labeling may be direct or indirect, and examples of direct labeling include recordings on physical objects such as the product itself, packaging, containers, labels, and signs, and examples of indirect labeling include advertising/promotional activities conducted through websites, stores, brochures, exhibitions, media seminars, books, newspapers, magazines, television, radio, mailings, e-mails, audio, and other venues or means.

The present invention is further described in detail below using examples, but the protection scope of the present invention is not limited to these examples.

[Example]

[Sugar solution preparation]

2’-FL (Jennewein Biotechnologie GmbH.), 3’-SL (Carbosynth), 6’-SL (Tokyo Chemical Industry #S0886), lactose (Fujifilm Wako Pure Chemical Industries, Ltd., #128-00095) and Neu5Ac (Fujifilm Wako Pure Chemical Industries, Ltd., #011-26173) were dissolved in sterile water to a concentration of 10% by mass and then sterilized using a 0.22 μm filter (Merck Millipore, #0369).

[Effects of human milk oligosaccharides on intestinal flora in human fecal culture system]

The fecal frozen solution provided by 8 healthy adults was thawed in a 37°C water bath. 70 μL of each fecal thaw solution was added to 7 mL of semi-fluid medium (Nissui #05460) for GAM sugar decomposition and cultured at 37°C under anaerobic conditions for 24 hours. 900 μL of each fecal culture solution was dispensed into 6 wells of a 96-well deepwell plate. 100 μL of each sugar solution (5 kinds) or sterilized water was added to the 6 wells (the final concentration of the sugar solution was 1% by mass). After culturing for 24 hours at 37°C under anaerobic conditions, DNA was extracted using Maxwell (company name: Promega) and amplicon sequencing of the V3-V4 region was performed using MiSeq (company name: Illumina). The obtained .fastq file was analyzed using QIIME2 (https://qiime2.org/) to calculate the occupancy of each intestinal bacteria.

The results are shown in Figure 1. In the groups given 2'-FL, 3'-SL, 6'-SL, lactose, and Neu5Ac, the occupancy rate of Collinsella bacteria increased significantly or tended to increase compared to the sterile water group as a control. This result indicates that 2'-FL, 3'-SL, 6'-SL, lactose, and Neu5Ac can promote the proliferation of Collinsella bacteria in the human intestine. It should be noted that Bifidobacterium bacteria were detected in all fecal thaw solutions.

In this example, lactose was confirmed to have a growth promoting effect on bacteria of the genus Collins. It is believed that HMO contains lactose as a constituent sugar, and that lactose is generated in the human digestive tract after HMO is ingested (Non-Patent Document 6). Therefore, it is believed that HMO can achieve a growth promoting effect on bacteria of the genus Collins.

In this example, Neu5Ac was confirmed to have a Collinsella bacteria growth promoting effect. It is believed that HMO containing Neu5Ac as a constituent sugar generates Neu5Ac in the human digestive tract after ingestion (Non-patent Documents 6 to 8). Therefore, it is believed that HMO (sialylated HMO, HMO containing sialic acid) containing Neu5Ac as a constituent sugar can obtain a Collinsella bacteria growth promoting effect.

In this example, the growth promoting effect on bacteria of the genus Collinsia was confirmed for 2'-FL, 3'-SL, and 6'-SL. From this, it is considered that the growth promoting effect on bacteria of the genus Collinsia can be obtained if it is fucosylated HMO or sialylated HMO.

[Papers cited in Examples]

Non-patent literature 6: Masi AC, Stewart CJ. Untangling human milk oligosaccharides and infant gut microbiome. iScience. 2021; 25(1): 103542. Published 2021Dec 1. doi: 10.1016/j.isci.2021.103542

Non-patent document 7: Nishiyama K, Yamamoto Y, Sugiyama M, et al. Bifidobacteriumbifidum Extracellular Sialidase Enhances Adhesion to the Mucosal Surface andSupports Carbohydrate Assimilation.mBio.2017;8(5):e00928-17.Published 2017Oct3.doi:10.1128/ mBio.00928-17

Non-patent literature 8: Kiyohara M, Tanigawa K, Chaiwangsri T, Katayama T, Ashida H, Yamamoto K. An exo-alpha-sialidase from bifidobacteria involved in the degradation of sialyloligosaccharides in human milk and intestinal glycoconjugates. Glycobiology. 2011; 21(4 ):437-447.doi:10.1093/glycob/cwq175

Claims (10)

1. A composition for suppressing the growth of bacteria belonging to the genus Collinsella in intestinal flora, the composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides. 2. The composition according to claim 1, comprising any one selected from the group consisting of fucosylated human milk oligosaccharides, sialylated human milk oligosaccharides, human milk oligosaccharides containing lactose as a constituent sugar, human milk oligosaccharides containing sialic acid as a constituent sugar, lactose and sialic acid. 3. The composition according to claim 2, comprising any one selected from the group consisting of 2'-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, lactose and N-acetylneuraminic acid. 4. The composition according to any one of claims 1 to 3, for use as a prebiotic or synbiotic. 5. A composition for treating a disease or condition improved by controlling the growth of Collins bacteria in the intestine, the composition comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides. 6. The composition according to claim 5, wherein the disease or condition improved by controlling the growth of Collins bacteria in the intestinal tract is a respiratory infection. 7. A composition for treating a respiratory infection, comprising any one selected from the group consisting of human milk oligosaccharides and constituent sugars of human milk oligosaccharides. 8. The composition according to claim 6 or 7, wherein the respiratory infection is a coronavirus infection. 9. The composition according to claim 6 or 7, wherein the respiratory infection is a novel coronavirus infection, i.e., COVID-19. 10 . The composition according to claim 1 , wherein the proliferation control is proliferation promotion.