TW201936204A - Composition for preventing or relieving respiratory disease - Google Patents

Abstract

Provided is a highly safe composition which is to be administered to pregnant and/or breastfeeding women so as to prevent a respiratory disease in infants born to the women. The composition for preventing a respiratory disease comprises a bacterium belonging to the genus Bifidobacterium as an active ingredient. The composition is to be administered to pregnant and/or breastfeeding women so as to prevent a respiratory disease in infants born to the women. This composition is particularly suitable for lowering the risk of the respiratory disease in the infants when the women are exposed to air pollution and/or passive smoking and thus the aforesaid risk is increased.

Description

Composition for prevention or alleviation of respiratory diseases

This technology relates to a composition for the prevention or alleviation of respiratory diseases.

It is known that the living environment of pregnant women may have an impact on the risk of respiratory diseases in children after birth. For example, it has been reported that a pregnant mother's exposure to air pollution causes an increase in asthma of a child born (Non-Patent Document 1). In addition, it has been reported that a child born to a mother who has second-hand smoke in pregnancy is significantly more likely to have asthma than a child born to a mother who does not have second-hand smoke (Non-Patent Document 2). For this reason, pregnant women should preferably live in an air-clean environment as much as possible, taking into account the risk of future fetal respiratory diseases.

In addition, means have been developed to correspond to the risk of disease after birth in the stage of pregnant women. For example, Patent Document 1 listed below discloses a prophylactic agent for allergic diseases in infants and young children, characterized in that Bifidobacterium bacterium is administered to a pregnant woman and a child thereof.
[Previous Technical Literature]
[Patent Literature]

Patent Document 1: Japanese Laid-Open Patent Publication No. 2014-65669.
[Non-patent literature]

Non-Patent Document 1: K. Hamada et al., J. Toxicol. Environ. Health A., 2007, Vol. 70(8), pp. 688-695.
Non-Patent Document 2: A. Zacharasiewicz, ERJ Open Res., 2016, Vol.2(3).

[Problems to be solved by the invention]

As mentioned above, although it is better for pregnant women to live in an air-clean environment as much as possible, it is often not easy to change the living environment. In particular, environmental factors such as exposure to air pollution and exposure to secondhand smoke are often difficult.
Therefore, the purpose of the present technology is to provide a new means for responding to the risk of respiratory diseases in children.
[Means for solving the problem]

As a result of research by the present inventors, it has been found that by administering Bifidobacterium bacteria to the mother during pregnancy and/or lactation, the risk of respiratory diseases in infants and young children can be met. That is, the present technology provides the following.

[1] A composition for preventing or slowing respiratory disease, comprising Bifidobacterium bacteria as an active ingredient.
The composition is administered to a mother during pregnancy and/or lactation and is used to prevent or slow the respiratory disease of an infant born to the mother.
[2] The composition according to the above [1], wherein the respiratory disease is a respiratory disease caused by a type 2 congenital lymphocyte.
[3] The composition according to the above [1] or [2] wherein the disease is caused or worsened by the presence of the parent in an air polluted environment.
[4] The composition according to any one of [1] to [3] wherein the respiratory disease is asthma.
[5] The composition according to any one of [1] to [4] wherein the respiratory disease is steroid-resistant asthma.
[6] The composition according to any one of [1] to [5] wherein the Bifidobacterium bacterium is Bifidobacterium breve.
[7] The composition according to any one of [1] to [6] wherein the Bifidobacterium bacterium is Bifidobacterium breve M-16V (NITE BP-02622).
[8] The composition according to any one of [1] to [7] wherein the composition is a drink or a food composition.
[9] The composition according to any one of [1] to [7] wherein the composition is a pharmaceutical composition.
[Effects of invention]

By the present technique, a Bifidobacterium bacterium is administered to a mother during pregnancy and/or lactation to prevent or slow down respiratory diseases of an infant born to the mother.
In addition, the effects of the present technology are not limited to the effects described in the present specification, and any of the effects described in the present specification may also be used.

Further preferred embodiments of the present technology will be described. However, the present technology is not limited to the following preferred embodiments, and can be freely changed within the scope of the present technology. In addition, the percentages of the present specification are expressed as mass unless otherwise specified.

<Composition for prevention or alleviation of respiratory diseases>
The composition for preventing or slowing down the respiratory disease of the present technology contains Bacillus bacteria as an active ingredient. The composition is administered to the mother during pregnancy and/or lactation and is used to prevent or slow the respiratory disease of the infant born to the mother.

According to the respiratory disease prevention composition of the present technology, respiratory diseases of infants and young children born to the mother can be prevented. The prevalence of respiratory diseases in children born to the mother who is unable to avoid living in an air polluted environment and who cannot avoid exposure to secondhand smoke, especially during pregnancy and/or breastfeeding, can be effectively reduced.
Further, the composition for preventing or slowing down the respiratory disease of the present technology can exert the preventive or slowing action as long as it is administered to the mother during pregnancy and/or lactation. Therefore, the composition for preventing or slowing down the respiratory disease of the present technology may not be administered to the child born to the mother. The administration of the composition of infants and young children is usually laborious. Since the composition of the present technology can be administered to a child born to the mother, it can be easily used.
The efficacy of the present technology is believed to be caused by the mother's ingestion of the Bifidobacterium bacteria during pregnancy and/or lactation, which causes changes in the mother's intestinal flora and/or breast milk, and the change in the constitution of the child born to the child. Come to influence and play.
The effects of the mother's intestinal flora and/or breast milk on the physical condition of the child born, such as the following report. Intestinal bacteria reported in pregnant mothers are associated with methylation of the child's diabetes-related genes (Tachibana K et al. J Diabetes Investig. 2017; 8(4): 550-553.). This is thought to be due to the fact that metabolites produced by the mother's intestinal bacteria pass through the placenta and affect epigenetic control during fetal development. It is also known that when the vagina is born, part of the mother's intestinal flora is taken over by the child. The intestinal flora of the mother who affects the intestinal flora of the infant is known to affect the child's future obesity, diabetes, nonalcoholic fatty liver disease, allergic diseases, etc. (Mulligan CM, Friedman JE, J Endocrinol. 2017;235(1): R1-R12.). In addition, the mother's breast milk is known to contain ingredients that prevent allergic diseases in children. By ingesting probiotics, lactating women change the concentration of ingredients related to allergy prevention in breast milk, suggesting that the probiotic intake of lactating mothers affects the child's constitution through breast milk (Munblit D et al. Clin Exp Allergy. 2015; 45(3): 583-601.). As mentioned above, the mother's intestinal flora and/or breast milk have an impact on the physical condition of the child born. The present technology also recognizes that the mother's intestinal flora and/or breast milk affects the physical condition of the child born, and as a result, prevents or slows down the respiratory disease.

The term "prevention" in the present technology includes preventing the onset of a disease, reducing the risk of the disease, reducing the likelihood of the onset of the disease, preventing the occurrence of at least one symptom of the disease, reducing the risk of occurrence of at least one symptom of the disease, and Reduce the likelihood of symptoms of at least one of the diseases.

The "mitigation" in the present technology includes the following: reducing the degree of the disease, delaying the progress of the disease, preventing the progress of the disease, alleviating the symptoms of at least one of the diseases, delaying the progress of the symptoms of at least one of the diseases, and preventing at least one of the diseases. Symptoms. In the case of respiratory diseases which are alleviated by the present technology, infants born from the mother or infants born from the mother may not be afflicted at the time of administration of the composition of the present technology to the mother. The composition of the present technology can alleviate the respiratory disease by the ingestion of the mother during pregnancy and/or lactation, in the case of a child born from the mother or a child born from the mother in the future. That is, the composition of the present technology can also be used to alleviate such respiratory diseases in the future.

In the present technology, the respiratory disease is preferably a respiratory disease caused by a Group 2 innate lymphoid cell (hereinafter referred to as "ILC2"). The compositions of the present technology are particularly useful for preventing or slowing down the respiratory disease. The respiratory disease of the present technology may be, for example, asthma, bronchitis, or pneumonia caused by type 2 congenital lymphocytes, and particularly may be asthma caused by type 2 congenital lymphocytes. As the asthma caused by the type 2 congenital lymphocytes, for example, steroid-resistant asthma can be cited.

The respiratory disease of the present technology may also be a disease accompanied by, for example, hyperplasia of airway epithelial goblet cells and/or increased eosinophilia in the bronchi. The compositions of the present technology are particularly useful for preventing or slowing down such diseases.
Examples of the respiratory diseases in the present technology include asthma, bronchitis, environmental lung disease, chronic bronchitis, COPD (Chronic Obstructive Pulmonary Disease), hypersensitivity pneumonitis, and eosinophilic pneumonia. However, it is not limited to this.

Further, in the present technology, the aforementioned diseases may be caused by the onset or deterioration of the aforementioned mother body in an air polluted environment. That is, it may be due to the above-mentioned maternal body in an air polluted environment, which may be the cause or deterioration factor of the aforementioned disease, or may be due to the presence of the parent in an air polluted environment, and other factors (such as hereditary factors and physical factors). And dietary factors, etc.) are the pathogenic factors or worsening factors of the aforementioned diseases.

In the present technology, the aforementioned precursor may also be in an air polluted environment. That is, the composition of the present technology can prevent and/or alleviate respiratory diseases of infants and young children born to the mother by administering the mother in an air-contaminated environment. In the present technology, the parent body is in an air polluted environment, and the mother body is stored in an air polluted environment. The aforementioned parent may also be in an air pollution environment during any period or all of the gestation period, during any period or all of the lactation period, or during the whole period of pregnancy and lactation, or for pregnancy Living in an air polluted environment before the period and not living in an air polluted environment during pregnancy and lactation. Further, the composition may be administered in a state where the precursor is exposed to air pollution, or may be administered in a clean space (for example, a clean room or the like) in an air polluted environment.

<Bifidobacterium bacteria>
The Bifidobacterium bacterium used in the present technology is not particularly limited as long as it does not impair the efficacy of the present technology, and a known Bifidobacterium bacterium can be used.
Examples of the Bifidobacterium bacterium include Bifidobacterium breve, Bifidobacterium longum subsp. longum, Bifidobacterium bifidum, and Bifidobacterium bifidum. Bifidobacterium adolescentis, Bifidobacterium angulatum, Bifidobacterium dentium, Bifidobacterium pseudocatenulatum, Bifidobacterium animalis subsp. lactis, Bifidobacterium pseudolongum subsp. globosum, Bifidobacterium pseudolongum subsp. pseudolongum, and Bifidobacterium thermophilum. The Bifidobacterium bacterium which is used in the present invention may be one or two or more selected from the above group, and one type may be used alone or two or more types may be used. In addition, the Rhizoctonia solani subsp. Rhizopus sp. is sometimes also abbreviated as Bifidobacterium breve.

Further, among the above Bifidobacterium bacteria, Bifidobacterium breve is preferred, and Bifidobacterium breve M-16V (NITE BP-02622) is more preferred.
Bifidobacterium breve M-16V (NITE BP-02622) was issued on January 26, 2018 in the Licensed Microbiology Depository Center of the Independent Administrative Corporation Product Evaluation Technology Base Agency (postal code: 292-0818, address: 122 , 2-5-8 Kazusa Kamatari Kisarazu-shi, Chiba JAPAN) International deposits under the Budapest Treaty, available from the deposit center.

As the Bifidobacterium bacterium used in the present technology, a mutant of the above Bifidobacterium bacterium can be used as long as it has the property of achieving the purpose of cost technology. Such a mutant can also be constructed by introducing a non-human mutation into the above Bifidobacterium bacteria. Further, it may be constructed by introducing a mutation into the above-mentioned bacteria by treatment using a mutant such as UV, or may be constructed by introducing a mutation into the above-described bacteria by various genetic manipulation methods.
In the case where the above mutant is used as a Bifidobacterium bacterium in the art, the mutant preferably has the same bacteriological properties as the Bifidobacterium bacterium described above, and has the same or higher than the Bifidobacterium bacterium described above. The inhibition of the production of ILC2. Whether or not a mutant has an effect of inhibiting the production of ILC2 by the Bifidobacterium bacterium is equivalent or less, and can be confirmed, for example, by measuring the ratio of ILC2 in the lung tissue by the method described in the test examples described later.

The Bifidobacterium bacterium used in the present technique is easily obtained by culturing a Bifidobacterium bacterium by a conventional method. Further, the Bifidobacterium bacterium of the present technology may use the aforementioned bacteria itself (bacteria), a culture of the aforementioned bacteria, a purified product derived from a culture of the aforementioned bacteria (for example, a culture supernatant), or a bacterium of the foregoing bacteria. The morphology of the body treatment.
That is, it may be a Bifidobacterium bacterium itself, or a culture obtained by culturing a Bifidobacterium bacterium may be used as it is. In addition, the Bifidobacterium bacteria used in the present technology may be either a live fungus or a dead fungus, or may contain both a living bacteria and a dead bacteria.

Examples of the bacterial cell-treated material of the above-mentioned bacteria include cell walls of the cells and cell disruptions in which the cell membrane is partially or completely broken by a conventional method such as ultrasonic treatment or homogenizer treatment. Further, the cell disruption may be a block in which the whole block after the rupture may be a part, the supernatant after the rupture is centrifuged, and the supernatant is partially purified by ammonium sulfate treatment, Or concentrating the above supernatant may also be used.

The Bifidobacterium bacterium used in the present technology has an effect of suppressing the excessive production of ILC2 in the lung tissue of an infant born to a baby by ingestion during pregnancy or lactation as shown in the examples below. That is, a mother who is pregnant or breast-feeding can ingest a Bifidobacterium bacterium according to the present technology, thereby effectively preventing or slowing a respiratory disease caused by ILC2 in an infant born to a child. Further, ILC2 in the lung tissue can be measured by a method as described in the examples below.

Here, ILC2 refers to a type 2 innate lymphoid cell of lymphocytes, which is activated by airway epithelial cells stimulated by air pollutants, microorganisms, glycolipids, etc. , producing Th2 cytokines (IL-5, IL-13). The aforementioned Th2 cytokine triggers activation of eosinophils or hyperplasia of goblet cells, and symptoms of asthma appear. The proliferation of goblet cells leads to overproduction of mucin in airway mucus, which is associated with airway remodeling. Airway remodeling exacerbates the symptoms of chronic respiratory diseases (Jantina C. de Groot et al., ERJ Open Res., 2015, Vol. 1(1)). The compositions of the present technology can also be used to prevent exacerbations such as chronic respiratory diseases. In addition, the compositions of the present technology can also be used to prevent or slow airway remodeling. In addition, the compositions of the present technology can also be used to prevent or slow the overproduction of mucin.

In the case of asthma with overproduction of ILC2, since symptoms are caused by not passing through Th2 cells, prevention or treatment of respiratory diseases such as the control of Th2 cells is not effective in some cases. According to the present technology, since the production of ILC2 can be suppressed, it is effective for the prevention or treatment of respiratory diseases which cannot be effectively prevented by the prevention or treatment of the activity control of Th2 cells.

Furthermore, activation or proliferation of ILC2 has been confirmed to be a cause of steroid-resistant asthma (H. Kabata et al, Nat. Commun., 2013, 4: 2675). Steroid-resistant asthma is refractory and is a serious disease for inhalation requiring high doses of steroids. In addition, steroid agents used in the treatment of asthma are known to have poor effects on asthma caused by pregnant women who smoke, and are prone to refractory asthma (Robyn T. Cohen et al., J. Allergy Clin Immunol., 2010) , 126(3), 491-497).
Since the composition for preventing or slowing down the respiratory disease of the present technology can suppress the overproduction of ILC2, it is effective even for refractory asthma prevention for steroid resistance in respiratory diseases.

The Bifidobacterium bacteria used in the present technology can be contained in the composition as an active ingredient, and can be used for a wide range of products such as pharmaceuticals, drinks, foods, and feeds because of high safety. These products can be produced by a known production method suitable for each use by appropriately using any component suitable for each application.
The Bifidobacterium bacterium used in the present technology can be used as it is, or can be used in combination with a physiological carrier, a pharmaceutical product, a usual carrier which can be tolerated in a drink or food, or a diluent.
Further, the Bifidobacterium bacteria used in the present technology can be used for the production of these various preparations or various compositions and the like.

The Bifidobacterium bacteria used in the art can also be used to prevent or alleviate allergic diseases. That is, the Bifidobacterium bacterium is a composition for preventing or alleviating an allergic disease, which is administered to a mother during pregnancy and/or lactation and prevents or slows the respiratory disease of an infant born to the mother. The above composition may also be contained as an active ingredient for prevention or alleviation.

<the object of the composition and the period of the administration>
The composition for the prevention of respiratory disease prevention of the present technology is applied to a mother who is pregnant and/or breast-feeding, and is particularly applicable to a pregnancy that must be lived in an environment exposed to air pollution and/or second-hand smoke. The mother in and/or breastfeeding. That is, the respiratory disease prevention or mitigation composition of the present technology is particularly suitable for reducing the risk of the aforementioned respiratory diseases caused by the aforementioned exposure of the mother to an air polluted environment and/or secondhand smoke. .
The term "parent" as used in the art refers to a mammal that is pregnant and/or breast-feeding, and the mammal is preferably a human or a pet, more preferably a human.
Further, the intake of the composition for preventing or slowing down the respiratory disease of the present technology is preferably a certain period of the gestation period and/or a certain period of the continuous lactation period. In particular, the intake of the composition of the present technology is for a certain period of time during pregnancy and lactation. Preferably, the compositions of the present technology continue to be ingested at least during pregnancy, preferably during pregnancy and during lactation. The composition of the present technology is preferably ingested the next day.
For example, the composition for preventing or slowing the respiratory disease of the present technology is calculated from at least 6 days before the expected date of birth, more preferably 4 weeks ago, and more preferably 2 months ago, 3 months ago, 4 months ago, 6 Ingested before the month of production, or 8 months before the date of production. In addition, the compositions of the present technology may also be taken during the entire period of pregnancy. Further, it is preferably ingested during the lactation period after birth, and is calculated from the production date of 30 days, more preferably 2 months, 3 months, 4 months, 6 months, or 8 months, and more preferably 1 year. Continue to ingest. In addition, the compositions of the present technology may also be ingested throughout the lactation period.

The composition for preventing or slowing down the respiratory disease of the present technology may also be administered only to the mother during pregnancy and/or lactation, without injecting or ingesting the infant born to the mother. The administration of the composition of infants and young children is usually laborious. Since the composition of the present technology can exert its effects without being administered to a child born to the mother, it can be easily used.

As described above, the respiratory disease prevention or mitigation composition of the present technology can reduce the exposure of the mother to the air by avoiding maternal administration in pregnancy and/or breastfeeding in an air polluted environment. The risk of the aforementioned respiratory diseases of the aforementioned infants and children who are raised in a polluted environment and/or in the environment of secondhand smoke.
Here, in the air pollution environment in the present specification, it means an environment in which air is contaminated by air pollutants such as fine particulate matter PM2.5. Specific examples of the environmental conditions of air pollution include the basis of the basic environmental law of Japan, which is based on the provisions of Article 16 (1) of the same law. With regard to the environmental conditions, in the same item, it is defined as a benchmark for maintaining the health of the person and maintaining the living environment. As a criterion for this, the air pollution is determined to be an environmental standard of an average annual concentration of fine particulate matter PM2.5 in air of 15 μg/m ³ or less and an average 24-hour concentration of 35 μg/m ³ or less. An environment exceeding this environmental standard can be exemplified as an air pollution environment in the present technology.

Further, the fine particulate matter PM2.5 refers to fine particles having a particle diameter of 2.5 μm or less suspended in the air. Examples of PM2.5 include gaseous air pollutants such as sulfuric oxides (SOx), nitrogen oxides (NOx), and volatile organic compounds (VOC) that are directly discharged by mass combustion, etc., mainly in ambient air. Particles are formed by chemical reactions. Examples of the source of PM2.5 include dust such as soot, coke ovens, and mineral accumulation fields such as boilers and incinerators, and exhaust gas such as automobiles, ships, and airplanes. Furthermore, PM2.5 is also produced from the natural world such as soil, ocean and volcano. In addition, cigarette smoke is also typical of PM2.5.
Further, in the animal model, when the PM2.5 is caused to cause health damage, as shown in the examples described later, the residual oil fly ash (ROFA) containing nickel, vanadium, and zinc as a main component can be used to reproduce the cause. Air pollution environment caused by PM2.5 (Non-Patent Document 1).

<medical composition>
The composition for preventing or slowing down the respiratory disease of the present technology can be used as a pharmaceutical composition. The pharmaceutical composition of the present technology can be used to prevent or slow down respiratory diseases of infants born to the mother by administering to the mother during pregnancy and/or breastfeeding. As respiratory diseases, bronchial asthma and steroid-resistant asthma can be exemplified.

In addition, the pharmaceutical composition of the present technology is a culture of a Bifidobacterium which is used for a long time as an oral composition component and/or a culture of the aforementioned bacteria (for example, a culture supernatant of the culture of the aforementioned bacteria and/or the aforementioned Since the bacterial cell treated product and the like are effective components, it can be administered to patients suffering from various diseases. Further, since Bifidobacterium bacteria are also present in the intestine of an animal, it is expected that the present technique is less likely to cause side effects even if administered for a long period of time and continuously. Therefore, the composition of the present technology can be safely taken even during pregnancy or breastfeeding.

When the composition for preventing or slowing the respiratory disease prevention of the present invention is used as a pharmaceutical composition, the pharmaceutical composition can be administered orally or parenterally, and can be formulated into an appropriate dosage according to the administration method. The required dosage form. For example, in the case of oral administration, it can be formulated into a solid preparation such as a powder, a granule, a tablet, or a capsule; a solution such as a solution, a syrup, a suspension, or an emulsion. Further, in the case of parenteral administration, for example, a composition of the present technology can be administered through a Percutaneous endoscopic gastrostomy.

Further, at the time of formulation, the pharmaceutical composition of the present technology may be a component such as an excipient, a pH adjuster, a coloring agent, or a flavoring agent which is usually used for formulation. Further, as long as the effects of the present technology are not impaired, the pharmaceutical composition of the present technology may also use an ingredient having a preventive, ameliorating, and/or therapeutic effect on a known or future discovered respiratory disease or a symptom thereof.
Further, the formulation can be carried out according to the dosage form by a suitably known method. At the time of formulation, the preparation carrier may be appropriately formulated and formulated.

The intake amount or the administration amount of the pharmaceutical composition of the present technology can be appropriately selected in accordance with the dosage form. For example, the intake or administration amount of the Bifidobacterium bacterium of 1 day per 1 kg of body weight is preferably 1 × 10 ⁵ CFU/ Kg body weight/day to 1×10 ¹² CFU/kg body weight/day, more preferably 1×10 ⁷ CFU/kg body weight/day to 1×10 ¹¹ CFU/kg body weight/day, and even more preferably 1×10 ⁸ CFU /kg body weight / day to 1 × 10 ¹⁰ CFU / kg body weight / day. Or, as the intake amount or administration amount per body weight, it is preferably 10 ⁶ CFU/day to 10 ¹⁴ CFU/day, more preferably 10 ⁸ CFU/day to 10 ¹³ CFU/day, and still more Good for 10 ⁹ CFU/day to 10 ¹² CFU/day.
Further, in the case of using a culture of the bacterium of the genus Bifidobacterium, a purified product (for example, a culture supernatant) of the culture of the bacterium, or a bacterial body treated product of the bacterium, the intake or the amount of the bacterium is preferably ingested or administered. When it is converted into the intake amount or the administration amount of the Bifidobacterium bacterium, the above-mentioned intake amount or administration amount is obtained.

Further, the pharmaceutical composition of the present technology is preferably ingested during pregnancy and/or lactation by the aforementioned intake amount or administration amount. In addition, at least from the 6th day before the expected date of birth, preferably better than 4 weeks ago, and more preferably 2 months ago, 3 months ago, 4 months ago, 6 months ago, or 8 months ago It is better to take up to the date of production. In addition, the pharmaceutical composition of the present technology can also be taken during the entire period of pregnancy. In addition, in the lactation period after birth, it is 30 days from the date of production, more preferably 2 months, 3 months, 4 months, 6 months, or 8 months, and more preferably 1 year. good. In addition, the pharmaceutical composition of the present technology can also be ingested throughout the lactation period.

Further, the content of the Bifidobacterium bacteria in the pharmaceutical composition of the present technology can be appropriately selected based on the aforementioned intake amount or administration amount. For example, it may be 1 × 10 ³ CFU / g to 1 × 10 ¹³ CFU / g or 1 × 10 ³ CFU / mL to 1 × 10 ¹³ CFU / mL, preferably 1 × 10 ⁷ CFU / g to 1 × 10 ¹¹ CFU/g or 1×10 ⁷ CFU/mL to 1×10 ¹¹ CFU/mL, more preferably 1×10 ⁸ CFU/g to 1×10 ¹⁰ CFU/g or 1×10 ⁸ CFU/mL to 1× 10 ¹⁰ CFU/mL. In the case where the bacterium is a dead bacterium, the CFU can be substituted for individual cells.
In addition, the content in the case of using the culture of the Bifidobacterium bacterium, the purified product derived from the culture of the bacterium (for example, the culture supernatant), or the bacterial cell treated product of the bacterium is preferably converted into In the case of the content of the Bifidobacterium bacterium, the content is the above content.

Further, as the aforementioned formulation carrier, various organic or inorganic carriers may be used depending on the dosage form. Examples of the carrier in the case of the solid preparation include an excipient, a binder, a disintegrator, a lubricant, a stabilizer, a flavoring and the like.

Examples of the excipient include sugar derivatives such as lactose, white sugar, glucose, mannite, and sorbit; starches such as corn starch, potato starch, α-starch, dextrin, and carboxymethyl starch. Derivatives; cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, carboxymethyl cellulose, carboxymethyl cellulose; gum arabic; dextran; pullulan; A phthalate derivative such as light anhydrous citric acid, synthetic aluminum ruthenate or aluminum bismuth citrate; a phosphate derivative such as calcium phosphate; a carbonate derivative such as calcium carbonate; a sulfate derivative such as calcium sulfate.

Examples of the binder include, in addition to the above excipients, gelatin, polyvinylpyrrolidone, polyethylene glycol, and the like.

The disintegrating agent may, for example, be a chemically modified starch or a cellulose derivative such as croscarmellose sodium, sodium carboxymethyl starch or crosslinked polyvinylpyrrolidone, in addition to the above-mentioned excipients.

Examples of the lubricant include talc; stearic acid; stearic acid metal salts such as calcium stearate and magnesium stearate; colloidal cerium oxide; glycerin aluminum citrate (VEEGUM); waxes such as cetyl wax; a diol; a carboxylic acid such as fumaric acid or adipic acid; a sodium salt of a carboxylic acid such as sodium benzoate; a sulfate such as sodium sulfate; leucine; a lauryl sulfate such as sodium lauryl sulfate or lauryl magnesium sulfate; An acid anhydride such as an acid anhydride or a citric acid hydrate; a starch derivative or the like.

Examples of the stabilizer include p-hydroxybenzoic acid esters such as methyl p-hydroxybenzoate and propyl p-hydroxybenzoate; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; dimethylammonium chloride; acetic anhydride; Sorbic acid, etc.

Examples of the flavoring and deodorizing agent include a sweetener, an acidulant, a flavor, and the like.
In addition, examples of the carrier used in the case of the liquid preparation for oral administration include a solvent such as water and a flavoring and odorizing agent.

<drinks, food composition>
Further, the composition for preventing or slowing down the respiratory disease of the present technology can be used as a drink or a food composition. The drink and food composition of the present technology may be a culture of a Bifidobacterium bacterium and/or the aforementioned bacteria (for example, a culture supernatant of a culture of the aforementioned bacteria and/or a bacterial body treatment of the aforementioned bacteria, etc.) It may be produced by adding to a known drink or food, and may also be a culture of Bifidobacterium and/or the aforementioned bacteria (for example, a culture supernatant of the culture of the aforementioned bacteria and/or a bacterial treatment of the aforementioned bacteria, etc.) It is produced as a new beverage or food composition by mixing it into raw materials of drinks and foods.

In addition to sugar lozenges, liquid foods, feeds (including pets), and the like, such as liquid, paste, solid, and powder, the beverages and foods of the present technology may, for example, be wheat flour products, instant foods, and agricultural products. Products, aquatic products, processed products, dairy products, dairy products, oils and fats, basic seasonings, compound seasonings (foods), frozen foods, desserts, beverages, and other commercial products.

In addition, the beverages and food compositions defined by the present technology can be supplied by the mother to be sold and sold as drinks, foods for the prevention of respiratory diseases, low risk, symptom reduction, and/or treatment (including health use) for indicating infants and young children. . In addition, as a target for the intake of drinks and foods, "pregnancy", "lacer", "the risk of respiratory diseases such as the fetus or infant", and "the risk of asthma or infants with asthma" can be indicated. Provided and sold by people who value the living environment during pregnancy or breastfeeding, and those who value the air in pregnancy or breastfeeding.
The "marking" behavior includes all the actions used to inform the demander of the above-mentioned uses. As long as it is reminiscent of, or analogy to, the performance of the above-mentioned uses, it is equivalent to the purpose of the label, the content of the label, the object of the label, the media, etc. The "marking" behavior of this technology.

In addition, the "marking" is preferably carried out by the demander directly recognizing the performance of the above-mentioned uses. Specifically, the transfer, delivery, transfer or delivery, display of the goods, or the packaging of the goods or products imported into the beverages, foods, or the goods described in the above-mentioned uses, are described in the advertisements, price lists, or transaction documents of the products. In addition, it is an act of providing an electromagnetic (network or the like) method by issuing or arranging the above-mentioned use as information in the content.

On the other hand, as the content of the label, it is preferable to be an approved label based on administration or the like (for example, approval based on various systems prescribed by the administration, marking based on such approval, etc.). In addition, such labeling content is preferably attached to promotional materials, other documents, etc. of the sales site such as packaging, containers, catalogs, manuals, POP (Point of Purchase).

In addition, the "marking" may also be used as a label for health foods, functional foods, enteral nutrition foods, special purpose foods, health functional foods, specific health foods, nutritional function foods, functional label foods, and quasi-drugs. In particular, the label recognized by the consumer institution may, for example, be a system for a specific health food, a nutritious food, or a functional food label, or a label recognized by a system similar thereto. Specific examples include, for example, a label for a specific health food, a label for a condition-specific health food, a label for an influence on a structure or function of the body, a disease risk reduction indicator, and a function based on a scientific basis; More specifically, the specific health foods prescribed by the Cabinet Office Order No. 57 of the Cabinet Office Order of August 31, 2011 on the licensing of special-purpose labels as prescribed by the Japanese Health Promotion Act. Markings (especially for health care purposes) and similar indications are typical examples.

The intake amount of the drink and food composition of the present technology can be appropriately selected. For example, the intake of Bifidobacterium bacteria per 1 kg of body weight is preferably 1 × 10 ⁵ CFU / kg body weight / day to 1 × 10 ¹² CFU. /kg body weight / day, more preferably 1 × 10 ⁷ CFU / kg body weight / day to 1 × 10 ¹¹ CFU / kg body weight / day, and more preferably 1 × 10 ⁸ CFU / kg body weight / day to 1 × 10 ¹⁰ CFU/kg body weight/day. Or, as the intake amount or administration amount per body weight, it is preferably 10 ⁶ CFU/day to 10 ¹⁴ CFU/day, more preferably 10 ⁸ CFU/day to 10 ¹³ CFU/day, and still more Good for 10 ⁹ CFU/day to 10 ¹² CFU/day. In the case where the bacterium is a dead bacterium, the CFU can be substituted for individual cells.
In addition, the intake amount in the case of using the culture of the Bifidobacterium bacterium or the bacterial cell treated product of the above-described bacteria is preferably the above-mentioned intake amount in the case of the intake amount of the Bifidobacterium bacterium.

Further, the beverage or food composition of the present technology is preferably ingested during pregnancy or breastfeeding in the above-mentioned intake or administration amount, from at least 6 days before the expected date of birth, more preferably 4 weeks ago, and even more preferably 2 Ingested before the month, 3 months ago, 4 months ago, 6 months ago, or 8 months ago. In addition, the beverages and food compositions of the present technology can also be ingested throughout the pregnancy period. Further, it is preferably ingested during the lactation period after birth, which is 30 days from the date of production, more preferably 2 months, 3 months, 4 months, 6 months, or 8 months, and more preferably 1 Ingestion during the year. In addition, the beverages and food compositions of the present technology can also be ingested throughout the lactation period.

Further, the content of the Bifidobacterium bacteria in the drink and food composition of the present technology can be appropriately selected based on the aforementioned intake amount, and may be, for example, 1 × 10 ⁶ CFU/g to 1 × 10 ¹² CFU/g or 1 ×. 10 ⁶ CFU/mL to 1×10 ¹² CFU/mL, preferably 1×10 ⁷ CFU/g to 1×10 ¹¹ CFU/g or 1×10 ⁷ CFU/mL to 1×10 ¹¹ CFU/mL, Preferably, it is 1 x 10 ⁸ CFU/g to 1 x 10 ¹⁰ CFU/g or 1 x 10 ⁸ CFU/mL to 1 x 10 ¹⁰ CFU/mL. In the case where the bacterium is a dead bacterium, the CFU can be substituted for individual cells.

The present technology can also provide the following methods and materials.
[1] A method of using a Bifidobacterium bacterium for the production of a composition for preventing or slowing a respiratory disease.
In the above method, the composition is administered to a mother during pregnancy and/or lactation, and the respiratory disease of the infant born to the mother is prevented or slowed down.
[2] A Bifidobacterium bacterium for preventing or slowing a respiratory disease.
Wherein the aforementioned Bifidobacterium bacterium is administered to a mother during pregnancy and/or lactation, and is used to prevent or slow down respiratory diseases of infants born to the mother.
[3] A method for preventing or slowing a respiratory disease of an infant born to the mother, comprising administering a Bifidobacterium bacterium to a mother during pregnancy and/or lactation.
[Examples]

[Test Example 1]
As a method for preparing a model of asthma caused by a child exposed to air-contaminated dust, a method of performing ROFA treatment on a pregnant female mouse and then performing egg white albumin treatment on the progeny mouse of the pregnant female mouse is established. (Non-Patent Document 1). In the test example 1, the asthma model was used to investigate the efficacy of administration of the parent Bifidobacterium bacterium.

The normal pregnant female mouse BALB/c (Japan SLC company) was divided into the following groups: Bifidobacterium bacteria M-16V (NITE BP-02622), Bifidobacterium bacterial ingestion group, no incurable Bifidobacterium A control group of bacteria, a non-treated group that did not ingest Bifidobacterium bacteria and did not receive the following ovalbumin treatment. In any of the groups, the ROFA infiltration solution 50 mg/ml was exposed for 30 minutes according to the method described in Non-Patent Document 1 on the 14th day, the 16th day, and the 18th day of pregnancy (before the 5th, 3rd, and the day before the birth). Aerosol. For each group of mice, they were allowed to freely ingest AIN93-G (Oriental Bio Service Co., Ltd.). In addition, in addition to the free intake, the control group and the non-treatment group ingested 0.5 g of AIN93-G refined feed once a day, and the ingestion group once ingested once in the AIN93-G refined feed 0.5 g, including 1 billion pairs. A feed of the genus Bacillus bacterium belonging to the genus Bacillus bacterium, M-16V (NITE BP-02622). The intake lasts until the end of the trial (between 6 days from the date of production and 30 days from the date of production).

The young mice born to the pregnant female mice in the ingestion group and the control group were intraperitoneally administered with 12.5 μg of ovalbumin (OVA, Sigma-Aldrich) and 12.5 μg of aluminum hydroxide on the 3rd day after birth. On the 26th, 27th, and 28th day, 1% of the OVA solution was exposed to a ultrasonic atomizer (PARI BOY N) for 10 minutes to prepare a respiratory disease model mouse with OVA as an antigen. In addition, egg albumin is a substance that causes respiratory diseases, and aluminum hydroxide is an adjuvant. Ovalbumin and aluminum hydroxide were administered in a state of being contained in a PBS solution. That is, 50 μl of a PBS solution containing ovalbumin at a concentration of 250 μg/ml and containing aluminum hydroxide at a concentration of 250 μg/ml was administered intraperitoneally.
The young mice born to the untreated group of pregnant female mice were intraperitoneally administered with 50 μl of the PBS solution containing the above-mentioned ovalbumin and aluminum hydroxide on the third day after birth, on the 26th, 27th and the following after birth. The PBS was exposed to a ultrasonic sprayer (PARI BOY N) for 10 minutes on the 28th.
All groups of young mice were dissected on the 30th day after birth by parenting with breast milk from the mother mice.

[The ratio of eosinophils in white blood cells]
After euthanasia in young mice, PBS was injected into the trachea to recover bronchoalveolarlavage fluid (BALF). After centrifugation of the bronchopulmonary cell washing solution and removal of the supernatant, the precipitate was resuspended in PBS, the total number of cells was counted, and a smear sample (using Cytospin: ThermoFisher Scientific) was prepared on a glass slide. Thereafter, Diff-Quik staining was performed using a Diff-Quik staining kit (Sysmex Corporation) and the white blood cell block in the bronchopulmonary cell wash was measured. The ratio of eosinophils in the white blood cells of each mouse was measured, and the average value of the eosinophilic ratio of each group was calculated.

As a result, the average value of the ratio of eosinophils in the white blood cells in the bronchopulmonary cell washing solution in each group was as shown in Table 1 and FIG.
The proportion of eosinophils in white blood cells was significantly higher in the control group than in the no treatment group. Therefore, in the control group, the eosinophils were significantly infiltrated into the bronchial tubes by the OVA treatment to form a respiratory disease.
In the ingested group, the proportion of eosinophils in the white blood cells was significantly lower than that in the control group, which was reduced by 12 or more. Therefore, the state of preventing or slowing down the respiratory disease by the uptake of the bacteria of the genus Bifidobacterium in the ingestion group.

[Table 1]
*1 has a significant difference (P<0.05)
*2 has a significant difference (P<0.05)

[Molecular fraction in lung tissue]
The lung tissues of the young mice were fixed with formalin and embedded into paraffin sections, and the mucins secreted by the goblet cells of the airway epithelium were stained by PAS staining. The histological evaluation of the stained sections and the degree of goblet cell hyperplasia of the airway epithelium were evaluated by the mucin fractions described in Table 2 below, based on C. Ptaschinski et al., PLOS Pathogens, DOI: The method described in 10.1371/journal.ppat.1004978, June 17, 2015 is carried out.

[Table 2]

As a result, the mucin fractions in the control group and the ingestion group are shown in Table 3 below and FIG. In the ingested group, the mucin fraction was significantly reduced compared to the control group. Therefore, goblet cell proliferation is inhibited by uptake of Bifidobacterium bacteria.

[table 3]
* Significant difference (P<0.05)

[ILC2 determination in lung tissue]
The lung tissue of the young mouse was separated into cell units, and the ratio of type 2 congenital lymphocytes (ILC2) was measured by flow cytometry MACSQuant Analyzer 10 (Miltenyi). To remove dead cells, staining was performed using 7-Amino-Actinomycin D (7-AAD). ILC2 is defined as Lineage(Lin)-CD45+, ST2(IL-33R)+, Thy1.2+ cells in the lymphocyte mass of FSC-SSC expansion, and the ratio of ILC2 in Lin-CD45+ cells is determined. The assay was carried out according to the method described in Duerr CU, Fritz JH. Methods Mol Biol. 2017; 1656: 253-261.
For lineage, antibodies containing CD3ε, CD4, CD5, CD8α, CD11c, CD19, F4/80, FcεRIα, Gr-1, NK1.1, and TER119 (all BioLegend) were used.

As a result, the ratio of ILC2 in the Lin-CD45+ cells in the control group and the ingestion group is shown in Table 4 and FIG. It was confirmed that the ratio of ILC2 was significantly reduced in the ingested group compared to the control group. Therefore, overproduction of ILC2 is inhibited by uptake of Bifidobacterium bacteria.

[Table 4]
* Significant difference (P<0.05)

As described above, by taking the Bifidobacterium bacterium in the mother during pregnancy or lactation, the mucin fraction of the larvae born and the ratio of the eosinophils can be reduced. Therefore, it can be seen that the respiratory disease of the larvae can be prevented or alleviated by the ingestion.
In addition, it has been confirmed that by ingesting Bifidobacterium bacteria during pregnancy or lactation, it is possible to suppress excessive production of ILC2 in the case of respiratory diseases caused by larvae of birth. In addition, the mucin fraction of the larvae and the proportion of eosinophils are reduced. From these results, it is known that the risk of developing respiratory diseases, especially respiratory diseases such as asthma, which are accompanied by overproduction of ILC2, is reduced.
As described above, by the present technique, it is possible to prevent or slow down respiratory diseases of infants born to children, especially respiratory diseases caused by overproduction of ILC2, by ingesting Bifidobacterium bacteria in the mother during pregnancy or breastfeeding. .

[Test Example 2]
As an experimental model of respiratory diseases caused by ILC2, a method of intranasal administration of the cytokine IL-33 to mice was established (Klein Wolterink RG, Eur J Immunol. 2012; 42(5): 1106-16 .). IL-33 airway epithelial cells are produced by stimulation with air pollutants or microorganisms. Intranasal administration of IL-33 is known to promote the production of Th2 cytokines by activation of ILC2 in lung tissue and to cause asthmatic symptoms.

In Test Example 2, the efficacy of the Bifidobacterium bacterium against the mother of the child was investigated with respect to the respiratory disease of the child caused by the ILC2.

In the same manner as in Test Example 1, normal pregnant female mice were divided into the following groups: a group in which Bifidobacterium bacteria M-16V (NITE BP-02622) was ingested, and a control group not ingested; Exposure by aerosol of ROFA infusion.

The young mice born to the aforementioned pregnant female mice were intranasally administered with 0.25 μg (BioLegend) on the 26th, 27th, and 28th day after birth. The young mice were bred on the 30th day of birth after being reared by the mother's breast milk nutrition.

The ratio of ILC2 in lung tissue activated by IL-33 was measured by flow cytometry. The measurement method was the same as the method described in Test Example 1.

As a result of the measurement, the ratio of ILC2 in the Lin-CD45+ cells in the control group and the ingestion group is shown in Table 5 below. The ratio of ILC2 was significantly reduced in the ingested group compared to the control group.

[table 5]
* Significant difference (P<0.05)

From the above results, it was confirmed that the parent acquired the Bifidobacterium bacterium during pregnancy and/or lactation, and it was possible to suppress the respiratory disease caused by the larvae of the child through the ILC2.

As shown in Test Example 2, the increase in ILC2 when the child was intranasally administered IL-33 was inhibited by the parent's intake of the Bifidobacterium bacteria during pregnancy and/or lactation. Therefore, by this ingestion, respiratory disease caused by permeation of ILC2 can be suppressed.

no.

Fig. 1 is a graph showing the average value of the proportion of eosinophils in white blood cells in the bronchopulmonary cell washing solution of the no treatment group, the control group, and the ingestion group.

Figure 2 is a graph showing the mucin scores of the control group and the ingested group.

Figure 3 is a graph showing the ratio of ILC2 in Lin-CD45+ cells of the control group and the ingested group.

Japan; the authorized microbial deposit center for product evaluation technology of the independent administrative agency (postal code: 292-0818, address: 122, 2-5-8 Kazusa Kamatari Kisarazu-shi, Chiba JAPAN); January 26, 2018; NITE BP -02622.

Claims (9)

A composition for preventing or slowing respiratory diseases, which comprises Bifidobacterium bacteria as an active ingredient; The composition is administered to a mother during pregnancy and/or lactation and is used to prevent or slow the respiratory disease of an infant born to the mother. The respiratory disease prevention or alleviation composition according to claim 1, wherein the respiratory disease is a respiratory disease caused by a type 2 congenital lymphocyte. The respiratory disease prevention or mitigation composition according to claim 1 or 2, wherein the disease is caused or worsened due to the presence of the mother in an air polluted environment. The respiratory disease prevention or alleviation composition according to claim 1 or 2, wherein the respiratory disease is asthma. The respiratory disease prevention or alleviation composition according to claim 1 or 2, wherein the respiratory disease is steroid-resistant asthma. The respiratory disease prevention or alleviation composition according to claim 1 or 2, wherein the Bifidobacterium bacterium is Bifidobacterium breve. The respiratory disease prevention or alleviation composition according to claim 1 or 2, wherein the Bifidobacterium bacterium is Bifidobacterium breve M-16V (NITE BP-02622). The respiratory disease prevention or alleviation composition according to claim 1 or 2, wherein the composition is a drink or a food composition. The respiratory disease prevention or alleviation composition according to claim 1 or 2, wherein the composition is a pharmaceutical composition.