JP7534870B2 - Composition for promoting intestinal development, and composition for preventing, treating, or alleviating symptoms of intestinal diseases - Google Patents

Description

NPMD NITE BP-02623

The present invention relates to a composition for promoting intestinal development and a composition for preventing, treating, or alleviating symptoms of intestinal diseases.

The digestive tract is a long tube that runs from the mouth to the anus, and is divided into the oral cavity, pharynx, esophagus, stomach, small intestine (duodenum, jejunum, ileum), and large intestine (cecum, appendix, ascending colon, transverse colon, descending colon, sigmoid colon, rectum). The intestinal tract, including the small intestine and large intestine, has the function of digesting food and absorbing nutrients. Normal development of the intestinal tract is important for the growth and maintenance of health of an individual.

It is known that some bifidobacteria and lactic acid bacteria improve the intestinal environment when ingested, thereby contributing to human health. For example, Patent Document 1 (JP Patent Publication No. 2014-111668) discloses that an oral composition containing Lactobacillus brevis and lactoferrin as active ingredients improves the frequency and amount of bowel movements.

JP 2014-111668 A

The present invention aims to provide a new composition that promotes intestinal development and a new composition that prevents, treats, or alleviates symptoms of intestinal diseases.

The present invention relates to the following examples [1] to [14].
[1] A composition for promoting intestinal development, comprising a bacterium belonging to Bifidobacterium longum subsp. infantis.
[2] The composition described in [1], which promotes intestinal fold formation.
[3] A composition for preventing, treating, or alleviating symptoms of an intestinal disease, comprising a bacterium belonging to Bifidobacterium longum subsp. infantis.
[4] The composition according to [3], wherein the intestinal disease is a disease caused by intestinal underdevelopment or intestinal dysfunction.
[5] The composition according to any one of [1] to [4], which is a food or drink composition.
[6] The composition according to any one of [1] to [5], wherein the bacterium is Bifidobacterium longum subsp. infantis M-63 (NITE BP-02623).
[7] The composition according to any one of [1] to [6], wherein the bacterium is a live bacterium.
[8] The composition according to any one of [1] to [7], further comprising human milk oligosaccharides.
[9] The composition according to any one of [1] to [8], which is a powdered milk preparation or a liquid milk preparation.
[10] An agent for promoting intestinal development, or an agent for preventing, treating, or alleviating symptoms of intestinal diseases, comprising a bacterium belonging to Bifidobacterium longum subsp. infantis.
[11] Use of a bacterium belonging to Bifidobacterium longum subsp. infantis for promoting intestinal development, or for preventing, treating, or alleviating symptoms of intestinal diseases.
[12] A bacterium belonging to Bifidobacterium longum subsp. infantis, used for promoting intestinal development, or for preventing, treating, or alleviating symptoms of intestinal diseases.
[13] Use of a bacterium belonging to Bifidobacterium longum subsp. infantis in the manufacture of a composition for promoting intestinal development, or a composition for preventing, treating, or alleviating symptoms of an intestinal disease.
[14] A method for promoting intestinal development, or a method for preventing, treating, or alleviating a symptom of an intestinal disease, comprising the step of administering to a mammal a bacterium belonging to Bifidobacterium longum subsp. infantis.

The present invention provides a new composition that promotes intestinal development and a new composition that prevents, treats, or alleviates symptoms of intestinal diseases.

1 is a photograph of the inner surface of the intestinal lumen of a pig in an M-63 administration group in an example. 1 is a photograph of the intestinal lumen surface of a pig in a control group in an example.

<Intestinal development promoter and composition for promoting intestinal development>
The intestinal development promoter according to the present invention includes a bacterium belonging to Bifidobacterium longum subsp. infantis. The intestinal development promoter according to the present invention can promote the development of the intestinal tract of a subject to which it is ingested or administered.

A composition according to one embodiment of the present invention is a composition for promoting intestinal development, and contains bacteria belonging to Bifidobacterium longum subsp. infantis. The composition for promoting intestinal development according to the present invention may contain the intestinal development promoter according to the present invention, or may be the intestinal development promoter according to the present invention itself. The composition for promoting intestinal development according to the present invention can promote intestinal development in a subject to which it is ingested or administered.

In this specification, the intestinal tract includes the small intestine and the large intestine. The intestinal tract is an organ responsible for food digestion and nutrient absorption. The small intestine includes the duodenum, jejunum, and ileum. Digestive enzymes are secreted from the mucous membrane layer of the small intestine, and food digested in the stomach is digested into final decomposition products such as amino acids, glucose, glycerides, and fatty acids. The small intestine repeatedly contracts and relaxes, moving the mixture of food and digestive juice while absorbing it. This movement helps to mix the digestive juice and food, and also increases the contact between the food and the mucosa, improving the absorption efficiency. The small intestine is composed of four layers from the inside: the mucosa, submucosa, muscularis, and serosal membrane. In the small intestine lumen, numerous folds (ring-shaped folds) are provided so as to surround the lumen, and countless villi exist on the surface of the folds, which contribute to increasing the surface area of the mucosa of the small intestine. The ring-shaped folds are composed of the mucosa and submucosa. The small intestine also functions as an immune organ, with aggregated lymphoid nodules (Peyer's patches) scattered on the inner surface of the small intestine. Major immune cells such as dendritic cells, T cells, and B cells are concentrated in Peyer's patches. These intestinal immune cell groups produce immunoglobulin A in response to invading pathogenic bacterial antigens, preventing the pathogenic bacteria from crossing the intestinal wall and entering the body.

The large intestine includes the cecum, appendix, ascending colon, transverse colon, descending colon, sigmoid colon and rectum. The large intestine absorbs water and salts, and moves the contents toward the rectum by peristalsis. The large intestine secretes mucus, and is composed of four layers from the inside: the mucosa, submucosa, muscularis and serosa. The large intestine has three taenia coli (taenia omenta, taenia mesocolon and free taenia) that run from the cecum to the sigmoid colon, and are composed of external longitudinal muscle. Contraction of the taenia coli forms folds (semi-lunar folds) in the lumen of the large intestine. A large number of intestinal bacteria live in the large intestine, which has a low oxygen concentration, and form the intestinal flora. It is known that the intestinal flora has a significant impact on the health of the host. Symptoms and diseases that are affected by the intestinal flora include constipation, diarrhea, irritable bowel syndrome, inflammatory bowel disease, metabolic diseases, infections, food poisoning, allergies, and malignant tumors such as colon cancer.

The development of the intestine means that the intestine has characteristics that allow it to better perform the intestinal functions described above. The development of the intestine includes the intestine having structural characteristics that allow it to better perform the intestinal functions of digesting food and absorbing nutrients. Specifically, the development of the intestine includes the formation of intestinal folds (especially the formation of circular folds in the small intestine), the formation of villi, the secretion of mucus, the development of smooth muscles (longitudinal muscles and circular muscles), the regulation of peristalsis, the secretion of gastrointestinal hormones, the development of the intrinsic nervous system that regulates peristalsis and hormone secretion, the secretion of digestive juices, the formation of intestinal flora, etc. These can affect each other. For example, when the formation of intestinal folds is insufficient, the surface area of the intestine decreases, and the number of cells present on the lumen side of the intestine also decreases, which can result in a decrease and underdevelopment of, for example, intestinal epithelial cells that absorb nutrients, goblet cells that secrete mucus, immune cells, intestinal bacteria, etc. Promoting intestinal development contributes to individual growth, weight gain, maintaining good nutritional status, promoting health, improving bowel movement frequency and volume, strengthening the barrier function of the intestinal wall, improving intestinal immune function, and preventing allergies, and may also have an effect on the brain, such as brain-gut correlation (reducing anxiety, improving cognitive function/suppressing decline).

In mammals, the nutritional supply system shifts from transplacental to enteral after birth. Newborns begin to ingest breast milk soon after birth, ingest nutrients from breast milk until weaning, and then switch to solid food after weaning. As this change in nutritional intake occurs, the morphology of the intestinal tract and intestinal mucosa, as well as the mechanism of nutrient digestion and absorption, change. Because infants' bodies are still immature, promoting intestinal development during infancy is important for their growth. Intestinal development is also important for infants whose immune systems are not yet fully functional, as it also helps improve immunity.

On the other hand, one of the causes of insufficient intestinal development is intestinal inflammation. In particular, when intestinal inflammation occurs in infants, it is thought that the formation of intestinal folds and villi is likely to be underdeveloped.

<Agent for preventing, treating or alleviating symptoms of intestinal disease, and composition for preventing, treating or alleviating symptoms of intestinal disease>
The agent for preventing, treating, or alleviating symptoms of intestinal diseases according to the present invention contains a bacterium belonging to Bifidobacterium longum subsp. infantis. The agent for preventing, treating, or alleviating symptoms of intestinal diseases according to the present invention can prevent, treat, or alleviate symptoms of intestinal diseases in a subject to which the agent is ingested or administered.

The composition according to one embodiment of the present invention is a composition for preventing, treating or alleviating symptoms of intestinal disease, and contains a bacterium belonging to Bifidobacterium longum subsp. infantis. The composition according to the present invention for preventing, treating or alleviating symptoms of intestinal disease may contain the agent for preventing, treating or alleviating symptoms of intestinal disease according to the present invention, or may be the agent for preventing, treating or alleviating symptoms of intestinal disease itself. The composition according to the present invention for preventing, treating or alleviating symptoms of intestinal disease can prevent, treat or alleviate symptoms of intestinal disease in a subject to which it is ingested or administered. Alleviating symptoms includes alleviating symptoms associated with the disease, delaying the progression of the disease, and shortening the time until recovery from the condition.

In this specification, intestinal diseases refer to diseases in the small intestine and large intestine. Examples of small intestinal diseases include, but are not limited to, small intestinal tumors, gastrointestinal bleeding, Crohn's disease, intussusception, gastrointestinal stromal tumors, malabsorption syndrome, Behcet's disease, etc. Examples of large intestinal diseases include, but are not limited to, colon cancer, polyps, colonic diverticulitis, acute appendicitis, irritable bowel syndrome, ulcerative colitis, ischemic colon, Crohn's disease, etc.

Bowel diseases are, for example, diseases caused by intestinal underdevelopment or decreased intestinal function, and include diseases that occur when the development or function of the intestine is insufficient as described above. Such bowel diseases include irritable bowel syndrome, inflammatory bowel disease, metabolic diseases, infectious diseases, food poisoning, allergies, malignant tumors such as colon cancer, and the like. Inflammatory bowel diseases include inflammation caused by external factors, such as microbial infections such as bacteria, trauma, physical stimuli such as heat, cold, radiation, and drugs, and chemical substances, as well as Crohn's disease, ulcerative colitis, necrotizing enterocolitis, Behçet's disease, and the like.

Preventing, treating or alleviating symptoms of intestinal diseases is expected to result in improved nutrient absorption in the intestinal tract, slower intestinal transit speed, increased appetite, weight gain, suppression of weight loss, prevention of allergic diseases and alleviation of symptoms, reduction of anxiety in the brain, and improvement/suppression of decline in cognitive function.

<Bacteria belonging to Bifidobacterium longum subsp. infantis>
Bacteria belonging to Bifidobacterium longum subsp. infantis are gram-positive obligately anaerobic rods. As the bacteria belonging to Bifidobacterium longum subsp. infantis, one or more types of bacteria belonging to known or unknown Bifidobacterium longum subsp. infantis can be arbitrarily selected as long as the effect of the present invention is not impaired. Bifidobacterium longum subsp. infantis may also be simply referred to as Bifidobacterium infantis. Examples of bacteria belonging to Bifidobacterium longum subsp. infantis include Bifidobacterium longum subsp. infantis BCCM LMG23728 and Bifidobacterium longum subsp. infantis M-63 (NITE BP-02623).

Bifidobacterium longum subsp. infantis BCCM LMG23728 is available from the Belgian Coordinating Collections of Microorganisms (BCCM), a Belgian collection, at 8, Scionstraat (Wetenskapstrasse), B-1000 Brussels, Belgium.

Bifidobacterium longum subsp. infantis M-63 (NITE BP-02623) was internationally deposited on January 26, 2018 at the National Institute of Technology and Evaluation Patent Microorganisms Deposit Center (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818) under the accession number NITE BP-02623 based on the Budapest Treaty. Bifidobacterium longum subsp. infantis BCCM LMG23728 and Bifidobacterium longum subsp. infantis M-63 (NITE BP-02623) are substantially the same bacterium.

Bifidobacterium longum subsp. infantis M-63 has a rod-shaped or branched pleomorphic shape, does not form spores, and is not motile. When the bacterium is spread on a BL agar medium plate and cultured at 37°C for 48 hours under anaerobic conditions, it forms opaque, circular, hemispherical, glossy colonies. Bifidobacterium longum subsp. infantis M-63 has the ability to ferment arabinose, xylose, glucose, fructose, galactose, sucrose, maltose, lactose, mevilose, raffinose, and turanose.

The bacteria belonging to Bifidobacterium longum subsp. infantis contained in the composition or agent according to the present invention may include bacteria substantially identical to the above-mentioned deposited strain. Examples of bacteria substantially identical to the deposited strain include derived strains having the deposited strain as a parent strain. Derived strains include strains bred from the deposited strain and strains that have naturally arisen from the deposited strain.

As used herein, "substantially the same bacterium" refers to a bacterium of the same genus or species, whose 16S rRNA gene base sequence has 98% or more, preferably 99% or more, and more preferably 100% identity with the 16S rRNA gene base sequence of the bacterium, and which preferably has the same bacteriological properties as the bacterium. Substantially identical bacteria are (1) bacteria determined to be the same strain by the RAPD (Randomly Amplified Polymorphic DNA) method or the PFGE (Pulsed-field gel electrophoresis) method (described in Probiotics in food/Health and nutritional properties and guidelines for evaluation 85, page 43), (2) bacteria that only have genes derived from the bacterial strain, have no genes of foreign origin, and have a DNA identity of 95% or more, or (3) strains bred from the bacterial strain (including genetic engineering modifications, mutations, and natural mutations, and may be bacteria with the same characteristics.

The bacteria belonging to Bifidobacterium longum subsp. infantis contained in the composition or agent according to the present invention may be live or dead, or may be both live and dead, but are preferably live. Microorganisms that are ingested or administered live and have a beneficial effect on the host are also called probiotics. The dead bacteria may be crushed. Additional procedures such as heating or freeze-drying may be performed after the culture, as long as they do not impair the effects of the present invention. It is preferable that the additional procedures are such that the survival of live bacteria is high.

The bacteria belonging to Bifidobacterium longum subsp. infantis contained in the composition or agent according to the present invention can be cultured by appropriately modifying the method usually used for culturing bacteria of the genus Bifidobacterium (bifidobacteria). The culture temperature may be, for example, 25 to 50°C, and preferably 30 to 40°C. The culture is preferably carried out under anaerobic conditions, and for example, the culture can be carried out while aerating anaerobic gas such as carbon dioxide. The pH of the medium is not particularly limited as long as it is within a range in which the bacteria can grow, and is, for example, pH 6.0 to 8.0.

The medium for culturing the bacteria belonging to Bifidobacterium longum subsp. infantis contained in the composition or agent according to the present invention is not particularly limited, and a medium usually used for culturing the bacteria can be used with appropriate modification as necessary. As the carbon source of the medium, for example, sugars such as galactose, glucose, fructose, mannose, cellobiose, maltose, lactose, sucrose, trehalose, starch, starch hydrolysate, and blackstrap molasses can be used depending on the assimilation ability. As the nitrogen source of the medium, for example, ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, and ammonium nitrate, and nitrates can be used. As the inorganic salt of the medium, for example, sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, and ferrous sulfate can be used. Organic components such as peptone, soybean flour, defatted soybean meal, meat extract, and yeast extract can also be used in the medium. As the prepared medium, for example, MRS medium can be preferably used.

Bacteria belonging to Bifidobacterium longum subsp. infantis can be used to promote intestinal development, or to prevent, treat, or alleviate symptoms of intestinal diseases. Bacteria belonging to Bifidobacterium longum subsp. infantis have been used in foods and medicines for many years, and exist as good bacteria in the intestines of animals, so they are expected to be highly safe for living organisms. In addition, they are thought to be unlikely to cause side effects or dependency, so they can be taken continuously for a long period of time, and can be taken easily by the target subjects.

The composition or agent of the present invention may contain bacteria other than those belonging to Bifidobacterium longum subsp. infantis, so long as it does not interfere with the intestinal development promoting effect or the intestinal disease prevention, treatment or symptom alleviation effect. Examples of bacteria that may be contained in the composition or agent of the present invention include other Bifidobacterium bacteria and lactic acid bacteria. Examples of other Bifidobacterium bacteria include Bifidobacterium bifidum, Bifidobacterium longum subsp. longum, Bifidobacterium adolescentis, Bifidobacterium pseudocatenulatum, Bifidobacterium animalis subsp. lactis, Bifidobacterium breve, Bifidobacterium pseudolonga, etc. Examples of lactic acid bacteria include Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus gasseri.

The composition or agent according to the present invention may contain a culture of bacteria belonging to Bifidobacterium longum subsp. infantis. The culture may be diluted or concentrated. The bacterial culture may contain secretions and metabolites of the bacteria.

<Food and drink composition>
The composition according to the present invention can be widely used as medicine, food and drink, and feed. The composition is preferably taken orally or through a feeding tube, more preferably taken orally. The composition can be taken daily in the form of medicine or food and drink. The composition according to the present invention is preferably a food and drink composition. The food and drink composition according to the present invention can safely and easily promote the development of the intestinal tract, or prevent, treat, or alleviate symptoms of intestinal diseases.

The food and drink composition according to the present invention may be in any form, such as liquid, paste, solid, or powder, and may also be in the form of tablet confectionery, liquid food, etc. Examples of the food and drink composition according to the present invention include flour products, instant foods, processed agricultural products, processed marine products, processed livestock products, milk and dairy products, oils and fats, basic seasonings, complex seasonings and foods, frozen foods, confectioneries, beverages, and other commercially available products.

Examples of flour products include bread, macaroni, spaghetti, noodles, cake mix, fried chicken flour, breadcrumbs, etc. Examples of instant foods include instant noodles, cup noodles, retort/prepared foods, pre-cooked foods, microwave foods, instant soups/stews, instant miso soup/cleaning soups, canned soups, freeze-dried foods, and other instant foods. Examples of processed agricultural products include canned agricultural products, canned fruit, jams/marmalades, pickles, boiled beans, dried agricultural products, and cereals (processed grain products). Examples of processed marine products include canned marine products, fish ham/sausages, marine paste products, marine delicacies, and tsukudani (food cooked in soy sauce). Examples of processed livestock products include canned livestock products/pastes, livestock ham/sausages, etc. Examples of milk and dairy products include fermented milk, processed milk, milk drinks, yogurts, lactic acid bacteria drinks, cheese, ice cream, modified milk powder, cream, and other dairy products. Examples of fats and oils include butter, margarines, and vegetable oils. Examples of basic seasonings include soy sauce, miso, sauces, tomato processed seasonings, mirin, and vinegars. Examples of complex seasonings and foods include cooking mixes, curry bases, sauces, dressings, noodle soups, spices, and other complex seasonings. Examples of frozen foods include ingredient frozen foods, semi-cooked frozen foods, and cooked frozen foods. Examples of confectioneries include caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice confectioneries, bean confectioneries, dessert confectioneries, and other confectioneries. Examples of beverages include carbonated beverages, natural fruit juices, fruit juice beverages, soft drinks with fruit juice, fruit pulp beverages, fruit beverages with fruit particles, vegetable beverages, soy milk, soy milk beverages, coffee beverages, tea beverages, powdered beverages, concentrated beverages, sports beverages, nutritional beverages, alcoholic beverages, and other beverages. Examples of commercially available foods other than those mentioned above include baby food, furikake, and ochazuke nori seaweed.

The content of the bacteria belonging to Bifidobacterium longum subsp. infantis in the food and drink composition is not particularly limited and can be appropriately selected based on the daily intake. The content of the bacteria belonging to Bifidobacterium longum subsp. infantis in the food and drink composition is, for example, preferably 1×10 ⁶ to 1×10 ¹² cfu/g or 1×10 ⁶ to 1×10 ¹² cfu/mL, more preferably 1×10 ⁷ to 1×10 ¹¹ cfu/g or 1×10 ⁷ to 1×10 ¹¹ cfu/mL, and even more preferably 1×10 ⁸ to 1×10 ¹⁰ cfu/g or 1×10 ⁸ to 1×10 ¹⁰ cfu/mL. The unit cfu is an abbreviation for colony forming units, and is a colony forming unit. If the bacteria are killed, cfu/g or cfu/mL can be replaced with cells/g or cells/mL.

The content of bacteria belonging to Bifidobacterium longum subsp. infantis in the food and beverage composition can be set at any amount as long as it does not impair the effects of the present invention. The content of bacteria belonging to Bifidobacterium longum subsp. infantis in the food and beverage composition can be set to 0.3% by mass to 5.0% by mass based on the final composition of the food and beverage composition.

The intake amount of the food or drink composition is preferably 1×10 to 1×10 cfu/day, more preferably 1× ¹⁰ to 1× ¹⁰ cfu/day, and even more preferably 1× ¹⁰ to 1× ¹⁰ cfu/day, as the amount of bacteria belonging to Bifidobacterium longum subsp. ^infantis per ^{kg of} body weight of the subject to be administered or ingested. When the bacteria are killed, cfu/day can be replaced with individual cells/day.

The food and drink composition may be a formula or liquid milk. Examples of formula include formula or liquid milk for infants, formula or liquid milk for follow-up, formula or liquid milk for low birth weight infants, formula or liquid milk for children, formula or liquid milk for adults, formula or liquid milk for the elderly, formula or liquid milk for allergies, formula or liquid milk for lactose intolerance, formula or liquid milk for congenital metabolic disorders, etc. It is particularly preferable that the food and drink composition is a formula or liquid milk for infants, a formula or liquid milk for follow-up, formula or liquid milk for low birth weight infants, or formula or liquid milk for children.

The composition may contain various additives depending on the type. The composition preferably contains human milk oligosaccharides. Human milk oligosaccharides is a general term for trisaccharide or higher oligosaccharides contained in human milk, excluding lactose. Mammalian milk has 12 core skeletons, such as lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-hexaose (LNH), and lacto-N-neohexaose (LNnH). By adding Lewis a, b, x, and α2-3/2-6N-acetylneuraminic acid to these skeletons, there are as many as 100 variations in glycans. Human milk oligosaccharides are contained in breast milk at 10 to 20 g/L, and may contain more than 130 types of oligosaccharides.

The human milk oligosaccharides contained in the composition may be one or more known or unknown human milk oligosaccharides that can be utilized by bacteria belonging to Bifidobacterium longum subsp. infantis, and may be selected from these oligosaccharides as desired, as long as they do not impair the effects of the present invention. Examples of human milk oligosaccharides include L-fucose, Lacto-N-fucopentaose I, Lacto-N-fucopentaose II, 2'-Fucosyllactose, 3'-Fucosyllactose, Lacto-N-fucopentaose III, 3'-Fucosyllacto-N-hexaose, Lacto-N-difucohexaose I, Lacto-difucotetraose, Lacto-difucohexaose I, and Lactodifucohexaose. II, Lacto-N-tetraose, Difucosyl syalyllacto-N-hexaose, Syalyllacto-N-tetraose, Fucosylsyalyl lacto-N-hexaose I, 3'-Fucosylsialy l lacto-N-tetraose (Fuc 1,2 Gal, Fuc 1,4 Gal NAC), Lacto-N-neotetraose, 3'-Sialyl-3-fucosyllactose, Disialomonofucosyllacto-N-neohexaose, Monofucosylmonosialyl lacto-N-oct aose, Sialylacto-N-fucohexaose II, Disialylacto-N-fucopentaose II, Monofucosyldisialyllacto-N-tetraose, 2'-Sialyllactose, 2'-Sialyllactosamine, 3'-Sialyllactose, 3'-Sialyllactosamine, 6'-Sia lyllactose, 6'-Sialylactosamine, Sialylacto-N-neotetraose c, Monosialylacto-N-hexaose, Disialylacto-N-hexaose I, Monosialylla cto-N-neohexaose I, Monosialyllacto-N-neohexaose II, Disialylacto-N-neohexaose, Disialylacto-N-tetraose, Disialylacto-N-hexaose II, Sialyllac to-N-tetraose a, Disialylacto-N-hexaose I, Sialylacto-N-tetraose b, 2'-Fucosyllacto-N-hexaose, 3'-Fucosyllacto-N-neohexaose, 2'-Fucosyl-N-acetylglucosamine, Lacto-N-fucopentaose V, Lacto-N-hex aose, Para-lacto-N-hexaose, Lacto-N-neohexaose, Para-lacto-N-neohexaose, Monofucosyllacto-N-hexaose II, Isomeric fucosylated lacto-N-hexaose (1), Isomeric fucosylated lacto-N-hexaose (3), Isomeric fucosylated lacto-N-hexaose (2), Difucosyl-para-lacto-N- neohexaose, Difucosyl-para-lacto-N-hexaose, 2' 3'-Difucosyllacto-N-hexaose, Lacto-N-neocataose, Para-lacto-N-octanose, Iso-lacto-N-octaose, Lacto-N-octaose, Monofucosyllact o-neoocataose, Monofucosyllacto-N-octaose, Difucosyllacto-N-octaose I, Difucosyllacto-N-octaose II, Difucosyllacto-N-octaose e II, Difucosyllacto-N-neooctaose I, Lacto-N-decaose, Trifucosyllacto-N-neooctaose, Trifucosyllacto-N-octaose, Trifucosyl-iso- Lacto-N-octaose, Lacto-N-difuco-hexaose II, Sialyl-lacto-N-tetraose a, Sialyl-lacto-N-tetraose b, Sialyl-lacto-N-tetraose c, Sialyl-fucosyl-lacto-N-tetraose I, Sialyl-fucosyl-lacto-N-tetraose II, Disialyl-lacto-N-tetraose, Lacto-N-neotetraose, etc., and combinations thereof. The composition of the present invention preferably contains at least one selected from 2'-fucosyllactose (hereinafter also referred to as "2'-FL"), lacto-N-fucopentaose I (hereinafter also referred to as "LNFPI"), lacto-N-difucohexaose I (hereinafter also referred to as "LNDFP I"), and lacto-N-tetraose (hereinafter also referred to as "LNnT").

2'-FL and LNnT are known to have bifidobacterial growth activity and act as bifidobacterial growth factors by being selectively utilized by bifidobacteria, and are important for the formation of a bifidobacterial-dominated intestinal flora. Human milk oligosaccharides can also be synthesized and used industrially.

The method of adding human milk oligosaccharides is not particularly limited as long as it does not impair the effects of the present invention. For example, human milk oligosaccharides can be added in liquid or powder form and mixed to produce the composition of the present invention.

The content of human milk oligosaccharides can be set at any amount as long as it does not impair the effects of the present invention. In the present invention, the content of human milk oligosaccharides in the composition can be set to 0.3% to 5.0% by mass based on the final composition.

The composition may further contain prebiotics. Prebiotics are indigestible food ingredients that have a beneficial effect on the host and improve the health of the host by selectively changing the growth and activity of specific bacteria in the large intestine. There are no particular limitations on the prebiotics as long as they do not impair the effects of the present invention. For example, lactulose, raffinose, galactooligosaccharides, fructooligosaccharides, soybean oligosaccharides, lactosaccharose, xylooligosaccharides, isomaltulose, coffee bean mannooligosaccharides, gluconic acid, polydextrose, inulin, etc. are preferred, and lactulose, raffinose, galactooligosaccharides, etc. are more preferred, with lactulose, raffinose and galactooligosaccharides being even more preferred.

The food and drink composition of the present invention can be produced by a method known to those skilled in the art, and may be produced by adding bacteria belonging to Bifidobacterium longum subsp. infantis to known foods and drinks, or by mixing the bacteria into the raw materials of foods and drinks to produce new food and drink compositions. The food composition can be produced by appropriately combining a molding process, a sterilization process, a fermentation process, a baking process, a drying process, a cooling process, a granulation process, a packaging process, and the like, in addition to the process of adding and mixing the bacteria belonging to Bifidobacterium longum subsp. infantis. The bacteria belonging to Bifidobacterium longum subsp. infantis may be used as a starter for producing fermented milk.

The content of the bacteria belonging to Bifidobacterium longum subsp. infantis in the production of the composition is not particularly limited, but is preferably 1× ¹⁰ to 1× ¹⁰ cfu/g or 1× ¹⁰ to 1× ¹⁰ cfu/mL, more preferably 1× ¹⁰ to 1× ¹⁰ cfu/g or 1× ¹⁰ to 1× ¹⁰ cfu/mL, and even more preferably ¹ ×10 to 1× ¹⁰ cfu/g or 1× ¹⁰ to 1× ¹⁰ cfu/mL. When the bacteria are killed, cfu/g or cfu/mL can be replaced with individual cells/g or individual cells/mL.

The food and drink composition for promoting intestinal development or the food and drink composition for preventing, treating or alleviating symptoms of intestinal disease can be provided and sold as a food and drink labeled with health uses such as intestinal development, prevention, treatment or alleviation of symptoms of intestinal disease. Such labels are not particularly limited, but examples include "promoting intestinal development," "improving nutrient absorption," "aiding digestion," "increasing weight," "promoting growth," "suppressing intestinal inflammation," "preventing intestinal disease," "preventing and treating infectious diseases," "preventing and treating food poisoning," "preventing and treating food allergies," "preventing and treating atopic dermatitis," "preventing and treating allergic rhinitis," "preventing and treating asthma," "preventing and treating anxiety symptoms," and "improving/suppressing decline in cognitive function." The act of "labeling" includes all acts for informing consumers of the above uses, and as long as the expression is capable of recalling or inferring the above uses, it all falls under the act of "labeling" of the present invention, regardless of the purpose of the labeling, the content of the labeling, the object or medium on which it is displayed, etc.

It is preferable that the content of the labeling be approved by the government (for example, a labeling approved based on various systems established by the government and made in a manner based on such approval). It is also preferable that such content of the labeling be affixed to promotional materials at the point of sale such as packaging, containers, catalogs, pamphlets, and POP, as well as other documents, etc.

"Labeling" also includes labeling as health food, functional food, enteral nutritional food, special purpose food, health functional food, food for specified health uses, nutrient functional food, functional food, medical drug, etc. Among these, labeling approved by the Consumer Affairs Agency, such as labeling approved under systems related to foods for specified health uses, nutrient functional foods, or foods with functional labels, or similar systems, can be mentioned. Specific examples include labeling as a food for specified health uses, labeling as a food for conditional specified health uses, labeling as a food with functional labels, labeling that indicates an effect on the structure or function of the body, labeling that reduces the risk of disease, and labeling of functionality based on scientific evidence.

<Pharmaceutical Composition>
The pharmaceutical composition is not particularly limited as long as it contains the bacteria belonging to Bifidobacterium longum subsp. infantis. The pharmaceutical composition may be produced by using the bacteria belonging to Bifidobacterium longum subsp. infantis as it is, or by mixing the bacteria with a physiologically acceptable liquid or pharmaceutical carrier.

The dosage form of the pharmaceutical composition is not particularly limited, and it can be formulated into solid preparations such as powders, granules, tablets, and capsules; liquid preparations such as solutions, syrups, suspensions, and emulsions; suppositories, ointments, and the like. When formulating the pharmaceutical composition, pharmaceutical carriers used in normal formulations can be used. The pharmaceutical composition can also contain a component that is known or will be discovered in the future to have an effect of promoting intestinal development or an effect of preventing, treating, or alleviating symptoms of intestinal diseases.

The content of the bacteria belonging to Bifidobacterium longum subsp. infantis in the pharmaceutical composition is not particularly limited, and can be appropriately selected based on the daily intake or dosage according to the dosage form. The content of the bacteria belonging to Bifidobacterium longum subsp. infantis in the pharmaceutical composition is, for example, preferably 1×10 ⁶ to 1×10 ¹² cfu/g or 1×10 ⁶ to 1×10 ¹² cfu/mL, more preferably 1×10 ⁷ to 1×10 ¹¹ cfu/g or 1×10 ⁷ to 1×10 ¹¹ cfu/mL, and even more preferably 1×10 ⁸ to 1×10 ¹⁰ cfu/g or 1×10 ⁸ to 1×10 ¹⁰ cfu/mL. If the bacteria are killed, cfu/g or cfu/mL can be replaced with cells/g or cells/mL.

The dosage of the pharmaceutical composition is preferably 1×10 ⁶ to 1×10 ¹² cfu/day, more preferably 1×10 ⁷ to 1×10 ¹¹ cfu/day, and even more preferably 1×10 ⁸ to 1×10 ¹⁰ cfu/day, as the amount of bacteria belonging to Bifidobacterium longum subsp. infantis per kg of body weight of the subject. When the bacteria are killed, cfu/day can be replaced with individual cells/day. The pharmaceutical composition may be administered orally or parenterally. The pharmaceutical composition may be administered once a day or in multiple doses.

As pharmaceutical carriers, various organic or inorganic carriers can be used depending on the dosage form. In the case of solid preparations, examples of carriers include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents, etc.

Examples of excipients include sugar derivatives such as lactose, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α-starch, dextrin, and carboxymethyl starch; cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, carboxymethyl cellulose, and calcium carboxymethyl cellulose; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; and sulfate derivatives such as calcium sulfate.

Examples of binders include the above excipients as well as gelatin, polyvinylpyrrolidone, and macrogol.

Disintegrants include, for example, the above-mentioned excipients as well as chemically modified starch or cellulose derivatives such as croscarmellose sodium, sodium carboxymethyl starch, and cross-linked polyvinylpyrrolidone.

Lubricants include, for example, talc; stearic acid; metal stearates such as calcium stearate and magnesium stearate; colloidal silica; waxes such as pea gum and galangal; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid; sodium carboxylates such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as silicic anhydride and silicic acid hydrate; and starch derivatives.

Examples of stabilizers include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; and sorbic acid.

Examples of flavoring agents include sweeteners, acidulants, and fragrances. In addition, examples of carriers used in liquid preparations for oral administration include solvents such as water, flavoring agents, etc.

The pharmaceutical composition may be administered alone or in combination with other medicines, such as other medicines for promoting intestinal development, or medicines for preventing, treating or alleviating symptoms of intestinal diseases, intestinal regulators, gastrointestinal agents, medicines for other intestinal diseases, painkillers, etc.

<Feed composition>
Examples of the feed composition according to the present invention include pet food, livestock feed, and fish feed. The feed composition can be produced by mixing bacteria belonging to Bifidobacterium longum subsp. infantis with general feed or its raw materials, such as grains, lees, bran, fish meal, bone meal, oils and fats, skim milk powder, whey, mineral feed, and yeast. For example, the feed composition may be produced through a fermentation process using bacteria belonging to Bifidobacterium longum subsp. infantis added, as in silage. The produced feed composition can be orally administered to general mammals, livestock, farmed fish, and pet animals.

The content of bacteria belonging to Bifidobacterium longum subsp. infantis in the feed composition is appropriately set depending on the form of the feed composition and the subject of administration, but is preferably 1× ¹⁰ to 1× ^{10 cfu/g or 1×10 to} 1× ¹⁰ cfu/mL, more preferably 1× ¹⁰ to 1× ¹⁰ cfu/g or 1× ¹⁰ to 1× ¹⁰ cfu/mL, and even more preferably 1× ¹⁰ to 1× ¹⁰ cfu/g or 1× ¹⁰ to 1× ¹⁰ cfu/mL. When the bacteria are killed, cfu/g or cfu/mL can be replaced with individual cells/g or individual cells/mL.

Methods for promoting intestinal development or preventing, treating, or alleviating symptoms of intestinal diseases
A method for promoting intestinal development or preventing, treating or alleviating symptoms of intestinal disease comprises administering to a mammal a bacterium belonging to Bifidobacterium longum subsp. infantis. The bacterium belonging to Bifidobacterium longum subsp. infantis may be administered in the form of a composition containing the bacterium belonging to Bifidobacterium longum subsp. infantis.

Mammals include humans, cows, sheep, goats, pigs, dogs, cats, horses, etc. The subjects of administration may be infants or human infants. Infants include infants and toddlers, more specifically infants, toddlers and newborns, more specifically infants, toddlers, newborns, premature infants, premature infants and low birth weight infants. Administration may be continued after infancy. In infants, administration of bacteria belonging to Bifidobacterium longum subsp. infantis facilitates proliferation of the bacteria in the intestine, and facilitates the promotion of intestinal development and the prevention, treatment or symptom relief of intestinal diseases.

In this specification, an infant refers to a child in infancy, and in the case of a human subject, infancy refers to the period from birth to less than one year of age (the day before the child turns one year old). Infancy refers to the period when milk, such as breast milk, is the main source of nutrition.

In this specification, the term "infant" refers to a child in the toddler stage, and in the case of a human subject, the toddler stage refers to the period from the day the child turns one year old until the day the child enters school age. The toddler stage preferably refers to the period from the day the child turns one year old until the day the child turns six years old or the day the child starts elementary school, whichever comes first.

When the subject is a human, the composition or agent of the present invention is preferably taken or administered from birth until before entering school age. When the subject is a human, the composition or agent of the present invention is more preferably taken or administered from birth until before the child reaches three years of age (until the day before the child reaches three years of age), even more preferably during infancy, and even more preferably from three months to less than one year of age (until the day before the child reaches one year of age).

When the subject is a mammal other than a human, the period may be converted into an equivalent period.
When the subject is a human, the "infancy period" corresponds to 20 days from birth in rats. Similarly, it corresponds to the day before the day when the subject becomes 10 weeks old from birth in cattle. It corresponds to the day before the day when the subject becomes 10 weeks old from birth in goats. It corresponds to the day before the day when the subject becomes 12 weeks old from birth in sheep. It corresponds to the day before the day when the subject becomes 8 weeks old from birth in pigs. It corresponds to the day before the day when the subject becomes 14 weeks old from birth in monkeys. It corresponds to the day before the day when the subject becomes 6 weeks old from birth in dogs. It corresponds to the day before the day when the subject becomes 6 weeks old from birth in cats. It corresponds to the day before the day when the subject becomes 20 days old from birth in mice. It corresponds to the day before the day when the subject becomes 20 days old from birth in hamsters. It corresponds to the day before the day when the subject becomes 20 days old from birth in guinea pigs.

When the subject is a human, the "infancy period" corresponds to the day before the day when the rat is 21 days old to 5 weeks old. Similarly, for cattle, it corresponds to the day before the day when the rat is 10 weeks old to 1 year old. For goats, it corresponds to the day before the day when the rat is 10 weeks old to 1 year old. For sheep, it corresponds to the day before the day when the rat is 12 weeks old to 1 year old. For pigs, it corresponds to the day before the day when the rat is 8 weeks old to 4 months old. For monkeys, it corresponds to the day before the day when the rat is 14 weeks old to 1.5 years old. For dogs, it corresponds to the day before the day when the rat is 6 weeks old to 6 months old. For cats, it corresponds to the day before the day when the rat is 6 weeks old to 6 months old. For mice, it corresponds to the day before the day when the rat is 21 days old to 5 weeks old. For hamsters, it corresponds to the day before the day when the rat is 21 days old to 5 weeks old. For guinea pigs, it corresponds to the day before the day when the rat is 21 days old to 5 weeks old.

When the subject is a human, "3 years of age" is equivalent to 4 weeks of age for rats. Similarly, for cows, it is equivalent to 6 months of age. For goats, it is equivalent to 6 months of age. For sheep, it is equivalent to 6 months of age. For pigs, it is equivalent to 3 months of age. For monkeys, it is equivalent to 9 months of age. For dogs, it is equivalent to 3 months of age. For cats, it is equivalent to 3 months of age. For mice, it is equivalent to 4 weeks of age. For hamsters, it is equivalent to 4 weeks of age. For guinea pigs, it is equivalent to 4 weeks of age.

When the subject is a human, "three full months" is equivalent to five full days of age for rats. Similarly, for cows, it is equivalent to 2.5 full weeks of age. For goats, it is equivalent to 2.5 full weeks of age. For sheep, it is equivalent to three full weeks of age. For pigs, it is equivalent to two full weeks of age. For monkeys, it is equivalent to three full weeks of age. For dogs, it is equivalent to 1.5 full weeks of age. For cats, it is equivalent to 1.5 full weeks of age. For mice, it is equivalent to five full days of age. For hamsters, it is equivalent to five full days of age. For guinea pigs, it is equivalent to five full days of age.

When the subject is a human, "one year of age" is equivalent to 21 days of age for rats. Similarly, for cows, it is equivalent to 10 weeks of age. For goats, it is equivalent to 10 weeks of age. For sheep, it is equivalent to 12 weeks of age. For pigs, it is equivalent to 8 weeks of age. For monkeys, it is equivalent to 14 weeks of age. For dogs, it is equivalent to 6 weeks of age. For cats, it is equivalent to 6 weeks of age. For mice, it is equivalent to 21 days of age. For hamsters, it is equivalent to 21 days of age. For guinea pigs, it is equivalent to 21 days of age.

The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these.

Five castrated male pigs of the LWD strain were used for each experimental group and were raised in a pig house at the Faculty of Food Industry, Miyagi University. The pigs were introduced at 4 weeks of age, acclimatized for 1 week, and then the test was started at 5 weeks of age. As the basal feed without added antibacterial agents, "HP Challenge High Milk One R", "Hachimantai A Mash", "Directly Produced Pig Fattening B" or "Directly Produced Pig Fattening C" from JA Zennoh Kitanihon Kumiai Feed Co., Ltd. were used according to the growth period. The pigs in the M-63 administration group were orally administered a suspension of Bifidobacterium longum subsp. infantis M-63 powder (hereinafter referred to as "M-63") in sterile PBS (-) once a day at 5 to 16 weeks of age. The dose of M-63 was 1.0 × 10 ¹¹ cfu/60 kg/day, and was increased stepwise every week according to the increase in body weight. The pigs in the control group were administered a suspension of tapioca starch in sterile PBS(-) in an amount equal to that of M-63 powder instead of M-63 powder.

At 19 weeks of age, the pigs were dissected and the digestive tracts were removed. Photographs of the intestinal lumen surface of the pigs in the M-63 administration group and the control group are shown in Figures 1 and 2, respectively. The small and large intestines of the pigs in the M-63 administration group had an increased number of folds compared to the pigs in the control group, and intestinal development was promoted. It is believed that intestinal diseases, particularly diseases caused by intestinal underdevelopment or reduced function, are prevented, treated, or symptoms are alleviated in the pigs in the M-63 administration group compared to the pigs in the control group.

The number of apexes of the circular folds per 10 cm of cross section in the longitudinal direction of the small intestine was counted to determine the average number of circular folds/cm. The number of apexes of the circular folds was counted at three locations per pig. The results are shown in Table 1. The number of circular folds was significantly increased in the pigs in the M-63 administration group compared to the pigs in the control group.

The average body weights of pigs in the M-63-administered and control groups at 15 weeks of age are shown in Table 2. Pigs in the M-63-administered group tended to gain weight better than pigs in the control group.

Claims (6)

A composition for promoting intestinal development , comprising Bifidobacterium longum subsp. infantis M-63 (NITE BP-02623) , wherein the intestinal development is intestinal fold formation . The composition according to claim 1 , which is a food or beverage composition. The composition according to claim 1 or 2 , wherein the Bifidobacterium longum subsp. infantis M-63 (NITE BP-02623) is a live bacterium. The composition according to any one of claims 1 to 3 , further comprising human milk oligosaccharides. The composition according to any one of claims 1 to 4 , which is a powdered milk formula or a liquid milk formula. The composition according to any one of claims 1 to 5, wherein the subject to take the composition is an infant.