Mucociliary clearance enhancers and foods and beverages for enhancing mucociliary clearance

Abstract

To provide agents and compositions for improving mucociliary clearance that can be easily taken orally as food and beverage, and to provide foods and beverages containing such compositions.SOLUTION: The above problems have been solved by using Lactococcus lactic acid bacteria as an active ingredient in a mucociliary clearance enhancer or a food or beverage composition for enhancing mucociliary clearance. In particular, the above problems have been solved by using a lactic acid bacterium belonging to Lactococcus lactis subsp. lactis, with accession number NITE BP-01694 at the National Patent Microorganisms Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE), as an active ingredient in a mucociliary clearance enhancer or a food or beverage composition for enhancing mucociliary clearance.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a mucociliary clearance enhancer containing a specific genus of lactic acid bacteria, a food and drink composition for enhancing mucociliary clearance, and a food and drink composition for enhancing mucociliary clearance that contains the food and drink composition for enhancing mucociliary clearance. [Background technology]

[0002] The epithelial surface area of ​​the airways from the nasal cavity to the trachea and alveoli is 140m 2 As a result, we are constantly exposed to substances that have a negative impact on the body, such as pathogens such as viruses and bacteria that enter through exhaled breath, harmful air pollutants such as PM2.5, and dust and pollen that can be allergens. To protect the airways from these foreign substances that enter the body through exhaled breath and prevent their entry into the body, the body has physical, chemical, and immunological defense mechanisms. The first of these defense mechanisms is mucociliary clearance, which is classified as a physical barrier. Because mucociliary clearance acts nonspecifically, it is the most important defense mechanism that can respond to foreign substances in general.

[0003] The majority of the surface of healthy airways is composed of ciliated and goblet cells and is covered by airway fluid, which consists of two layers: an upper mucus layer (gel layer) that traps inhaled particles and pathogens, and a low-viscosity periciliary layer (sol layer) that lubricates the airway surface and promotes efficient ciliary beating. Foreign particles trapped by the mucus layer are transported toward the mouth by ciliary beating at a speed of 4–20 mm per minute and are ultimately eliminated by expectoration or swallowing. Therefore, a decrease in mucociliary clearance may lead to an increased risk of infection and the progression of respiratory diseases due to the retention of air pollutants and allergens (Non-Patent Documents 1 and 2).

[0004] Mucociliary clearance is known to decrease due to dryness and a drop in environmental temperature (Non-Patent Document 3). This is thought to be related to influenza epidemics in winter. It is also known that mucociliary clearance decreases with aging due to weakening of ciliary beats and changes in mucus composition (Non-Patent Document 2). This suggests that elderly people are at a higher risk of developing respiratory infections due to delayed elimination of pathogens during early bacterial or viral infections (Non-Patent Documents 2 and 4). Activating mucociliary clearance and rapidly eliminating foreign substances that enter the airways through exhalation or aspiration is expected to reduce the risk of respiratory diseases caused by infectious diseases, allergies, and harmful pollutants.

[0005] Various reports have been published regarding agents for improving mucociliary clearance and methods for improving mucociliary clearance.

[0006] Patent Document 1 discloses a ciliary movement activator containing sodium bicarbonate.

[0007] Patent Document 2 discloses an airway foreign body removal agent comprising 2-tridecen-1-ol, tetrahydrogeraniol, dupical, isocyclemon, citral dimethyl acetal, acetophenone, carvone, hexylcyclopentanone, menthofuran, bourbonene, β-methyl-decalactone, muscone, isoeugenol, γ-dodecalactone, or γ-hexadecalactone.

[0008] Patent Document 3 discloses a method for treating mucosal diseases and disorders by administering an effective amount of poly(acetyl,arginyl)glucosamine (PAAG) having a specific structure to improve mucociliary transport and mucociliary clearance.

[0009] Patent Document 4 discloses a method for treating a disease that is improved by increasing mucociliary clearance and mucosal hydration, which comprises administering an osmolyte and a sodium channel blocker with a specific structure to a subject in need of increased mucociliary clearance and mucosal hydration.

[0010] Patent Document 5 discloses a method for promoting the rate of mucociliary clearance in a subject in need of such treatment, which comprises administering to the subject an effective mucociliary clearance-stimulating amount of a composition comprising a Kunitz-type serine protease inhibitor and a physiologically acceptable carrier.

[0011] The above-mentioned prior art techniques identify chemical substances that are presumed to be involved in improving mucociliary clearance, but in some cases specific agents for ingesting such chemical substances have not been put into practical use, and even if specific agents exist, they may not be easily ingested, leaving room for further improvement. [Prior art documents] [Patent documents]

[0012] [Patent Document 1] Patent Publication No. 2021-102606 [Patent Document 2] Japanese Patent Application Laid-Open No. 2010-260855 [Patent Document 3] Special Publication No. 2017-526714 [Patent Document 4] Special Publication No. 2010-502738 [Patent Document 5] Special Publication No. 2002-532558 [Patent Document 6] Patent No. 6505018 [Non-patent literature]

[0013] [Non-Patent Document 1] Int Arch Otorhinolaryngol. 2021; 25(1):e35-e40. [Non-patent document 2] Adv Geriatr Med Res. 2022; 4(2): e220005. [Non-patent document 3] Proc Natl Acad Sci US A. 2019;116(22):10905-10910. [Non-patent document 4] Am J Respir Crit Care Med. 2001;163(4):983-8. [Non-Patent Document 5] Arch Environ Health. 1974; 29(5):290-3. [Non-patent document 6] Br J Dis Chest. 1984; 78(1):62-5. Summary of the Invention [Problem to be solved by the invention]

[0014] The present invention has been made in view of the above-mentioned background art, and an object of the present invention is to provide an agent or composition for improving mucociliary clearance that can be easily taken orally as a food or beverage, and a food or beverage containing such a composition. [Means for solving the problem]

[0015] The present inventors have conducted extensive research to solve the above problems and have discovered the following facts, which have led to the completion of the present invention.

[0016] That is, the present inventors have found that oral ingestion of lactic acid bacteria improves (enhances) mucociliary clearance in individuals who take the bacteria. Lactic acid bacteria are currently used as probiotics in a variety of foods and beverages, and mucociliary clearance enhancers containing lactic acid bacteria as active ingredients can be added to a variety of foods and beverages and easily ingested.

[0017] The present invention thus completed is as follows:

[0018] A mucociliary clearance enhancer characterized by containing Lactococcus lactic acid bacteria as an active ingredient.

[0019] A food and drink composition for enhancing mucociliary clearance, characterized in that it contains Lactococcus lactic acid bacteria as an active ingredient.

[0020] A food or drink for enhancing mucociliary clearance, comprising the food or drink composition for enhancing mucociliary clearance. [Effects of the Invention]

[0021] According to the present invention, it is possible to provide an agent or composition for improving mucociliary clearance, which can be easily taken orally as a food or drink, and a food or drink containing such a composition.

[0022] Oral ingestion of the food and drink of the present invention can reduce the risk of respiratory diseases by promoting the elimination of harmful foreign substances from the environment through enhancement of mucociliary clearance, which is a major defense mechanism. [Brief explanation of the drawings]

[0023] [Figure 1] 1 is a graph showing the amount of fluorescent beads remaining in the trachea of ​​the 11 / 19-B1 lactic acid bacteria administration group and the control group, measured in Evaluation Example 1. [Figure 2] 1 is a graph showing the amount of fluorescent beads remaining in the lungs of the 11 / 19-B1 lactic acid bacteria administration group and the control group, measured in Evaluation Example 2. [Figure 3] 1 is a graph showing the amount of fluorescent beads remaining in the trachea of ​​the group administered lactic acid bacteria 11 / 19-B1 for a long period of time, the group administered lactic acid bacteria 11 / 19-B1 for a short period of time, and the control group, measured in Evaluation Example 3. [Figure 4] 1 is a graph showing the results of saccharin time measured in the 11 / 19-B1 intake group and the placebo group in Evaluation Example 4. DETAILED DESCRIPTION OF THE INVENTION

[0024] The present invention will be described below, but the present invention is not limited to the following embodiments and can be practiced with any modifications.

[0025] <Mucociliary clearance enhancer> The present invention relates to a mucociliary clearance enhancer characterized by containing Lactococcus lactic acid bacteria as an active ingredient.

[0026] Lactococcus lactic acid bacteria are gram-positive bacteria belonging to the family Streptococcus, and have been widely used in foods and beverages such as butter, cheese, and lactic acid bacteria drinks.

[0027] Examples of species of Lactococcus lactic acid bacteria that are active ingredients in the mucociliary clearance enhancer of the present invention include Lactococcus lactis, Lactococcus chungangensis, Lactococcus garvieae, Lactococcus piscium, Lactococcus plantarum, Lactococcus raffinolactis, and Lactococcus cremoris. Of these, Lactococcus lactis is particularly desirable as the active ingredient of the mucociliary clearance enhancer of the present invention because of its ease of availability and its extensive history of use in various foods and beverages.

[0028] There are two known subspecies of Lactococcus lactis: Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. hordniae, and either of these can be the active ingredient of the mucociliary clearance enhancer of the present invention. Lactococcus lactis subsp. lactis is particularly preferred as the active ingredient of the mucociliary clearance enhancer of the present invention.

[0029] In addition, the following can be exemplified as specific strains of Lactococcus lactis (Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. holdoniae).

[0030] <(A) Lactococcus lactis subsp. lactis> (A1) Lactic acid bacteria with accession number NITE BP-01694 at the National Patent Microorganism Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE). (A2) JCM5805 strain (A3) JCM1158 strain (A4) JCM7638 strain (A5) JCM12650 shares (A6) JCM20101 strain (A7)JCM20128 stock (A8) JCM20312 strain (A9) JCM20399 strain (A10) ATCC19435 strain (A11) ATCC15577 strain (A12) ATCC11454 strain (A13) ATCC7962 strain (A14) ATCC15346 strain (A15) ATCC29146 strain (A16) ATCC11007 strain (A17) ATCC baa-1055 strain (A18) ATCC13675 strain (A19) ATCC11955 strain (A20) ATCC7963 strain (A21) ATCC12929 strain (A22) ATCC27861 strain (A23)DSM20481 strain (A24)DSM20175 strain (A25)DSM4366 strain (A26)DSM20250 strain (A27)DSM20384 strain (A28)DSM20661 strain (A29)DSM20729 strain (A30)DSM27303 strain (A31)DSM30863 strain (A32)DSM110082 strain (A33)DSM111832 strain (A34)DSM111868 strain (A35)DSM111869 strain (A36)DSM111870 strain (A37)NBRC100933 strain (A38)NBRC12007 strain (A39)NBRC114714 strain (A40)NBRC114784 strain (A41)NRIC1149 strain (A42)NRIC1074 strain (A43) NRIC1150 stocks (A44)NRIC0497 strain (A45)NRIC1147 strain (A46)NRIC1148 strain

[0031] <(B) Lactococcus lactis subsp. holdoniae> (B1) JCM1180 strain (B2) JCM11040 strain (B3) ATCC29071 strain (B4) DSM20450 strain (B5)NBRC100931 strain

[0032] The lactic acid bacteria (A2) to (A9), (B1), and (B2) above are available from the Microbial Materials Development Division, BioResource Center, RIKEN. The above lactic acid bacteria (A10) to (A22) and (B3) are available from ATCC (American Type Culture Collection) (USA). The above lactic acid bacteria (A23) to (A36) and (B4) are available from DMSZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen) (Germany). The above lactic acid bacteria (A37) to (A40) and (B5) are available from the National Institute of Technology and Evaluation, Biotechnology Center. The above lactic acid bacteria (A41) to (A46) are available from the Microbial Resource Center, Tokyo University of Agriculture.

[0033] It should be noted that the same strain may be available from multiple institutions and may be assigned different numbers by each institution. Among the specific examples of Lactococcus lactis listed above, the same strain is listed twice with different numbers.

[0034] The lactic acid bacterium, with accession number NITE BP-01694 at the National Patent Microorganisms Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE), is a species of Lactococcus lactis subsp. lactis, and details thereof are described in Patent Document 6. Hereinafter, "the lactic acid bacterium with accession number NITE BP-01694 at the National Patent Microorganism Deposit Center (NPMD) of the National Institute of Technology and Evaluation (NITE)" may be referred to as "11 / 19-B1 strain" or "11 / 19-B1 lactic acid bacterium."

[0035] The 11 / 19-B1 strain is a microorganism that has been deposited domestically at the National Patent Microorganism Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE), 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan, under accession number NITE P-01694 (deposit date: August 20, 2013). The 11 / 19-B1 strain was then transferred from domestic deposit (original deposit date: August 20, 2013) to deposit under the Budapest Treaty (international deposit date: October 15, 2014) by submitting an original deposit application to the National Patent Microorganisms Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE), 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan, and was assigned the accession number "NITE BP-01694" after its viability was verified and the application for transfer to deposit under the Budapest Treaty (international deposit) was accepted.

[0036] The mucociliary clearance enhancer of the present invention can contain various Lactococcus lactic acid bacteria as active ingredients, as described above. In the mucociliary clearance enhancer of the present invention, the active ingredient may be one type of Lactococcus lactic acid bacteria, or two or more types of Lactococcus lactic acid bacteria.

[0037] The mucociliary clearance enhancer of the present invention can contain the active ingredient, Lactococcus lactic acid bacteria, in various states.

[0038] The mucociliary clearance enhancer of the present invention may contain Lactococcus lactic acid bacteria in the form of live bacteria or killed bacteria. For example, live bacteria, wet bacteria, or dried bacteria can be appropriately used as the active ingredient of the mucociliary clearance enhancer of the present invention. In addition, killed bacteria that have been subjected to heat sterilization, radiation sterilization, disruption, or the like can also be appropriately used as the active ingredient of the mucociliary clearance enhancer of the present invention.

[0039] The mucociliary clearance enhancer of the present invention may contain a treated product obtained by subjecting live or killed Lactococcus lactic acid bacteria to various treatments. Such processed products include, for example, cultures; concentrates; pastes; dried products such as spray-dried products, freeze-dried products, vacuum-dried products, and drum-dried products; liquefied products; diluted products; crushed products; sterilized products; and extracts from the cultures.

[0040] The content of Lactococcus lactic acid bacteria in the mucociliary clearance enhancer of the present invention (the total content of live bacteria, killed bacteria, and processed products; if two or more types of Lactococcus lactic acid bacteria are contained, the total content of live bacteria, killed bacteria, and processed products for all of the Lactococcus lactic acid bacteria) is not particularly limited and can be selected appropriately depending on the purpose. The content is equivalent to 1 x 10 4 Preferably, it is 1×10 or more. 5 More preferably, 1×10 6 It is particularly preferable that the number is 1×10 or more. 7 It is more preferable that the content is 1×10 or more. 13 Preferably, it is 1×10 or less. 12 It is more preferable that the number is 5×10 or less. 11 It is particularly preferable that the number is 1×10 or less. 11 It is more preferable that the number is 1 or less.

[0041] The mucociliary clearance enhancer of the present invention may contain "other ingredients" in addition to the Lactococcus lactic acid bacteria as the active ingredient. The "other components" are not particularly limited and can be appropriately selected depending on the purpose as long as the effects of the present invention are not impaired. For example, additives used in processing such as lyophilization and pharmaceutically acceptable carriers can be cited as "other components." Such a carrier is not particularly limited and can be appropriately selected depending on the dosage form, which will be described later.

[0042] The "other components" contained in the mucociliary clearance enhancer of the present invention include not only intentionally added components but also components produced by Lactococcus lactic acid bacteria, such as dextran.

[0043] There are no particular limitations on the amount of "other ingredients" contained in the mucociliary clearance enhancer of the present invention, and the amount can be selected appropriately depending on the purpose.

[0044] The dosage form of the mucociliary clearance enhancer of the present invention is not particularly limited and can be appropriately selected depending on the desired administration method, etc., as described below. Specific examples include oral solid preparations (tablets, coated tablets, granules, powders, capsules, etc.), oral liquid preparations (oral liquids, syrups, elixirs, etc.), injections (solvents, suspensions, etc.), ointments, patches, gels, creams, external powders, sprays, and inhalation powders.

[0045] The oral solid preparation can be produced by a conventional method, for example, by adding an excipient to the active ingredient, and further adding additives such as a binder, a disintegrant, a lubricant, a colorant, a flavoring agent, and an odorant, as needed. Examples of the excipient include lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, microcrystalline cellulose, and silicic acid. Examples of the binder include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl starch, methyl cellulose, ethyl cellulose, shellac, calcium phosphate, and polyvinylpyrrolidone. Examples of the disintegrants include dry starch, sodium alginate, agar powder, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, monoglyceride stearate, and lactose. Examples of the lubricant include purified talc, stearates, borax, and polyethylene glycol. Examples of the colorant include titanium oxide and iron oxide. Examples of the flavoring and / or odor correcting agent include sucrose, orange peel, citric acid, and tartaric acid.

[0046] The oral liquid preparation can be produced by adding additives such as flavoring agents, buffering agents, and stabilizers to the active ingredient in a conventional manner. Examples of the flavoring and / or odor correcting agent include sucrose, orange peel, citric acid, and tartaric acid. The buffering agent may be, for example, sodium citrate. Examples of the stabilizer include tragacanth, gum arabic, and gelatin.

[0047] As the injection, for example, a pH adjusting agent, a buffering agent, a stabilizer, an isotonic agent, a local anesthetic, etc. can be added to the active ingredient, and an injection for subcutaneous, intramuscular, intravenous, etc. can be produced by a conventional method. Examples of the pH adjuster and the buffer include sodium citrate, sodium acetate, and sodium phosphate. Examples of the stabilizer include sodium pyrosulfite, EDTA, thioglycolic acid, and thiolactic acid. Examples of the isotonic agent include sodium chloride and glucose. Examples of the local anesthetic include procaine hydrochloride and lidocaine hydrochloride.

[0048] The ointment can be prepared, for example, by combining the active ingredient with a known base, stabilizer, humectant, preservative, etc., and mixing them in a conventional manner. Examples of the base include liquid paraffin, white petrolatum, white beeswax, octyldodecyl alcohol, and paraffin. Examples of the preservative include methyl parahydroxybenzoate, ethyl parahydroxybenzoate, and propyl parahydroxybenzoate.

[0049] The patch can be produced, for example, by applying the ointment in the form of a cream, gel, paste, or the like to a known support in a conventional manner. Examples of the support include woven or nonwoven fabrics made of cotton, staple fiber, chemical fiber, etc.; and films or foam sheets made of soft vinyl chloride, polyethylene, polyurethane, etc.

[0050] The subjects to which the mucociliary clearance enhancer of the present invention may be administered are not particularly limited, and examples include humans; mice; rats; monkeys; horses; livestock such as cows, pigs, goats, sheep, and chickens; and pets such as cats, dogs, rabbits, and hamsters.

[0051] When the mucociliary clearance enhancer of the present invention is administered to humans, elderly people, smokers, and people who experience discomfort in the throat, i.e., people whose mucociliary clearance enhancer activity tends to be weakened, are particularly desirable as administration targets.

[0052] The method of administration of the mucociliary clearance enhancer of the present invention is not particularly limited and can be appropriately selected depending on the dosage form, etc. Specific examples include oral administration, intraperitoneal administration, injection into the blood, injection into the intestine, gargling, and nasal irrigation (nasal gargling).

[0053] The dosage of the mucociliary clearance enhancer of the present invention is not particularly limited and can be appropriately selected depending on the age and body weight of the individual to be administered, the degree of the desired effect, and the like. For example, the daily dose of the mucociliary clearance enhancer of the present invention to an adult is 1×10 6 Preferably, 1 x 10 7 More preferably, 1×10 8 It is particularly preferable that the dose is 1×10 or more. 13 Preferably, it is 1×10 or less. 12 It is more preferable that the number is 5×10 or less. 11 It is particularly preferred that the number is equal to or less than 100.

[0054] The timing of administration of the mucociliary clearance enhancer of the present invention is not particularly limited and can be appropriately selected depending on the purpose. For example, the administration may be prophylactic or therapeutic.

[0055] Furthermore, as mentioned above, it is known that mucociliary clearance is reduced by dryness and a drop in environmental temperature. Therefore, it is preferable to administer the mucociliary clearance enhancer of the present invention during times when dryness and a drop in temperature are likely, such as winter.

[0056] There are no particular limitations on the frequency of administration of the mucociliary clearance enhancer of the present invention. The administration frequency is preferably at least once every three days, more preferably at least once a day, and particularly preferably at least three times a day. The administration frequency is preferably 30 times or less a day, more preferably 20 times or less a day, and particularly preferably 10 times or less a day.

[0057] The mucociliary clearance enhancer of the present invention is more likely to exhibit sufficient effects when taken continuously. The mucociliary clearance enhancer of the present invention is preferably taken continuously for, for example, one week or more, two weeks or more, one month or more, three months or more, six months or more, or one year or more.

[0058] <Food and drink composition for enhancing mucociliary clearance> The present invention also relates to a food or drink composition for enhancing mucociliary clearance, which comprises Lactococcus lactic acid bacteria as an active ingredient. That is, the mucociliary clearance enhancer of the present invention can be used as an additive to foods and beverages. Details of the food and drink composition for enhancing mucociliary clearance of the present invention are the same as those of the mucociliary clearance enhancer described above.

[0059] <Food and drink for enhancing mucociliary clearance> The present invention also relates to a food or drink for enhancing mucociliary clearance, which contains the above-mentioned food or drink composition for enhancing mucociliary clearance.

[0060] The food and drink product for enhancing mucociliary clearance of the present invention may further contain "other ingredients" in addition to the above-described food and drink product composition for enhancing mucociliary clearance.

[0061] The "other components" in the food and beverage product for enhancing mucociliary clearance of the present invention are not particularly limited and can be appropriately selected depending on the purpose as long as the effects of the present invention are not impaired. Examples of the "other components" include various food ingredients. Furthermore, the content of the "other components" is not particularly limited and can be appropriately selected depending on the purpose.

[0062] The food and beverage products for enhancing mucociliary clearance of the present invention may be chilled products (products stored at a temperature of 0°C or higher and 10°C or lower) or non-chilled products (products stored frozen, products stored at room temperature, etc.).

[0063] The type of food or drink for enhancing mucociliary clearance of the present invention is not particularly limited and can be appropriately selected depending on the purpose. These include sweets such as jelly, candy, chocolate, biscuits, and ice cream; beverages such as green tea, black tea, coffee, and soft drinks; dairy products such as fermented milk (hard yogurt, soft yogurt, drink yogurt, frozen yogurt, etc.), lactic acid bacteria drinks, pudding, ice cream, ice milk, and lacto ice cream; processed vegetable and fruit products such as vegetable drinks, fruit drinks, and jams; liquid foods such as soups; processed grain products such as bread and noodles; various seasonings; foods for specified health uses; and foods with nutritional functions. Among these, dairy products such as fermented milk and lactic acid bacteria drinks are particularly preferred as foods and beverages for enhancing mucociliary clearance of the present invention.

[0064] The mucociliary clearance enhancing food and drink of the present invention may be produced, for example, as oral solid preparations such as tablets, granules, capsules, etc., or oral liquid preparations such as oral liquid preparations, syrups, etc. The method for producing the oral solid preparations and oral liquid preparations is not particularly limited and can be appropriately selected depending on the purpose. For example, they can be produced following the method for producing oral solid preparations and oral liquid preparations of the above-mentioned drugs (mucociliary clearance enhancers).

[0065] The content of Lactococcus lactic acid bacteria in the food and beverage products of the present invention for enhancing mucociliary clearance (the total content of live bacteria, killed bacteria, and processed products; if two or more types of Lactococcus lactic acid bacteria are contained, the total content of live bacteria, killed bacteria, and processed products for all Lactococcus lactic acid bacteria) is not particularly limited and can be selected appropriately depending on the purpose. The content is 1 x 10 per 100g of food and drink for enhancing mucociliary clearance. 6 Preferably, it is 1×10 or more. 7 More preferably, 1×10 8 It is particularly preferable that the number is 1×10 or more. 9 It is more preferable that the content is 1×10 or more per 100 g of the food or drink for enhancing mucociliary clearance. 13 Preferably, it is 1×10 or less. 12 It is more preferable that the number is 5×10 or less. 11 It is particularly preferable that the number is 1×10 or less. 11 It is more preferable that the number is 1 or less. [Example]

[0066] The present invention will be explained in more detail below by way of examples and comparative examples, but the present invention is not limited to these examples as long as it does not depart from the gist of the invention.

[0067] Evaluation example 1 (mucociliary clearance in the trachea) [Preparation of heat-killed bacteria] Colonies of 11 / 19-B1 lactic acid bacteria cultured on MRS agar medium were suspended in MRS liquid medium to prepare a bacterial suspension. 200 μL of the bacterial suspension was added to 200 mL of MRS liquid medium and then allowed to stand for 48 hours. After 48 hours of culture, the culture was centrifuged at 3000 × g for 20 minutes at 4°C, the medium was removed, and the bacterial pellet was suspended in 30 mL of ice-cold saline. The cells were centrifuged at 2900 × g for 20 minutes at 4°C. After centrifugation, the supernatant was discarded and the cells were suspended in 30 mL of milliQ water. The cells were then centrifuged again under the same conditions to wash them. The supernatant was discarded and the cells were suspended in 10 mL of ice-cold milliQ water ("milliQ" is a registered trademark). They were then immersed in boiling water for 10 minutes for heat sterilization, cooled on ice, frozen in a deep freezer, and subsequently lyophilized to obtain a heat-killed bacterial powder. Weigh the heat-killed bacterial powder and calculate the bacterial dry weight to bacterial count using a conversion table prepared in advance. 10 The cells were suspended in Dulbecco's phosphate buffered saline (D-PBS) to a concentration of 100 cells / mL to prepare a killed cell preparation, which was stored at -80°C until use.

[0068] [Administration of killed 11 / 19-B1 lactic acid bacteria to living mice] Female C57BL / 6J mice (Jackson Laboratory Japan) were used, and from the age of 8 weeks, 200 μL (2 × 10) of 11 / 19-B1 killed lactic acid bacteria suspension in D-PBS was administered. 9 The animals were orally administered with a sonde every day for two weeks. The control group was administered the same amount of D-PBS instead of the 11 / 19-B1 killed lactic acid bacteria fluid. Two weeks later, the final oral administration was given, and the evaluation test for tracheal mucociliary clearance began one hour after administration.

[0069] [Evaluation of tracheal mucociliary clearance] Under isoflurane anesthesia, mice were secured to the intubation table. While viewing through a mouse laryngoscope (Natsume Seisakusho), 5 μL of 25 mg / mL fluorescent silica beads, sicastar-greenF, φ100 nm (Corefront Co., Ltd.), was sprayed into the trachea using a spray sonde (24G / W103 mm, Natsume Seisakusho) with a 24G / W103 mm spray tube (Natsume Seisakusho). After spraying, the mice were slowly removed from the intubation table and allowed to recover from anesthesia. They were then housed under normal conditions for 2 hours. After 2 hours, the mice were dissected, and the main trachea from the subepiglottis to the bifurcation was collected and immersed in 500 μL of cold PBS. 500 μL of 4% TritonX100 / PBS was added to the tracheal immersion solution, and the trachea was then incised longitudinally. This was vortexed for 5 minutes, then left on ice for 5 minutes, and repeated three times to obtain tracheal lavage fluid. To remove tissue fragments and debris, the tracheal wash was centrifuged at 100 × g for 1 minute at 4°C (it was confirmed that fluorescent silica beads did not precipitate under these centrifugation conditions), and 100 μL of the supernatant was dispensed in duplicate into a 96-well fluorescence plate and measured using a plate reader (Excitation 485 nm, Emission 520 nm).

[0070] [result] The results of measuring the fluorescence intensity of the tracheal lavage fluid are shown in Figure 1. It was confirmed that the amount of fluorescent silica beads remaining in the trachea after spraying was significantly lower in the 11 / 19-B1 lactobacillus-treated group compared to the control group. This indicates that the rate of mucociliary clearance in the trachea was faster in the 11 / 19-B1 lactobacillus-treated group compared to the control group.

[0071] Evaluation example 2 (excretion of foreign matter from the lungs) [Preparation of heat-killed bacteria and administration of 11 / 19-B1 killed lactic acid bacteria to living mice] The same procedure as in Evaluation Example 1 was carried out.

[0072] [Evaluation of lung clearance] Under isoflurane anesthesia, mice were secured to an intubation table. While viewing through a mouse laryngoscope (Natsume Seisakusho Co., Ltd.), 5 μL of 25 mg / mL fluorescent silica beads (sicastar-greenF, φ100 nm, Corefront Co., Ltd.) was sprayed into the trachea using a spray sonde 24G / W103 mm (Natsume Seisakusho Co., Ltd.). After spraying, the mice were slowly removed from the intubation table and allowed to recover from anesthesia. They were then housed under normal conditions for 16 hours. After 16 hours, the mice were dissected and whole lungs were collected. A volume of 0.05% NP-40 substitute (nonylphenyl polyethylene glycol) in PBS was added to the lungs in an amount four times the lung weight, and the lungs were crushed using a biomasher to obtain lung extracts. 570 μL of 2% TritonX100 in PBS was added to 30 μL of lung extract and thoroughly mixed to prepare a suspension for measurement. 100 μL of the suspension for measurement was dispensed in duplicate into a 96-well fluorescence plate and measured using a plate reader (Excitation 485 nm, Emission 520 nm).

[0073] [result] The results of measuring the fluorescence intensity of the lung extract are shown in Figure 2. It was confirmed that the amount of fluorescent silica beads remaining in the lungs after passing through the trachea was significantly lower in the 11 / 19-B1-treated group compared to the control group. This indicates that the mucociliary clearance rate in the bronchioles in the 11 / 19-B1-treated group was faster than in the control group, resulting in more rapid expulsion of foreign matter from the lungs.

[0074] Evaluation example 3 (mucociliary clearance in the trachea) [Preparation of bacterial cells] A colony of 11 / 19-B1 lactic acid bacteria was smeared over the entire surface of an M17 agar medium with a cotton swab and cultured statically for 48 hours. After 48 hours of culture, the grown bacteria were collected with a cotton swab and suspended in 10 mL of D-PBS to prepare a bacterial cell preparation.

[0075] [Administration of 11 / 19-B1 lactic acid bacteria to living mice] The effect of administering 11 / 19-B1 lactic acid bacteria on the suppression of mucociliary clearance was evaluated using aged mice. Male C57BL / 6J mice (Jackson Laboratory Japan) were divided into three groups: a long-term administration group, a short-term administration group, and a control group. From 12 months of age, the control group and the short-term administration group received D-PBS, and the long-term administration group received 200 μL (1 × 10 9 The mice were orally administered 1 × 10 cfu of the above-mentioned bacterial cell preparation (200 μL) using a probe 3 days a week. Two weeks before the mucociliary clearance ability evaluation test at 21 months of age, the administration was changed to daily. The control group received D-PBS, and the short-term and long-term administration groups received 200 μL (1 × 10 cfu) of the above-mentioned bacterial cell preparation suspended in 1 × 10 cfu. 9 The test to evaluate the mucociliary clearance ability of the trachea was started one hour after the last oral administration.

[0076] [Evaluation of tracheal mucociliary clearance] Evaluation was carried out in the same manner as in Evaluation Example 1.

[0077] [result] The results of measuring the fluorescence intensity of the tracheal lavage fluid are shown in Figure 3. The amount of fluorescent silica beads remaining in the trachea decreased depending on the duration of administration of 11 / 19-B1 lactic acid bacteria. The amount remaining was significantly reduced in the long-term administration group compared to the control group, indicating that administration of 11 / 19-B1 lactic acid bacteria suppressed the decline in tracheal mucociliary clearance ability due to aging.

[0078] Evaluation Example 4 (Human Mucociliary Clearance) The mucociliary clearance ability of 11 / 19-B1 Lactobacillus in humans was evaluated using the saccharin test. The saccharin test measures the time (saccharin time) from when saccharin granules are attached to the nasal mucosa until they are transported to the pharynx by mucociliary clearance and are perceived as sweet. Although the saccharin test has the drawback of not directly measuring mucociliary clearance in the trachea and bronchi, previous reports have demonstrated a correlation between mucociliary transport function in the nasal cavity and the trachea (Non-Patent Document 5), and the test is used as a test method reflecting mucociliary clearance in the trachea and bronchi. Furthermore, it can be performed relatively easily and non-invasively, and its reproducibility within the same individual has been confirmed (Non-Patent Document 6).

[0079] [subject] Males and females aged 35 to 60 years (those who did not meet the exclusion criteria for health status, lifestyle habits, or medication intake) were selected as potential subjects. A preliminary saccharin test was conducted on 25 candidates, and 20 subjects with a slow saccharin time (decreased mucociliary movement) were enrolled as subjects. According to the protocol, the enrolled subjects were randomly assigned by the allocation manager before the study to either the test food intake group (10 subjects, 5 men, 5 women, mean age 46.3 years) or the placebo intake group (10 subjects, 4 men, 6 women, mean age 41.4 years).

[0080] [Test meal] The test food was a drink containing 11 / 19-B1 lactic acid bacteria (sterilized lactic acid bacteria drink 1 x 10 10 The test food was administered once a day (100 mL) for four weeks. A beverage containing no 11 / 19-B1 lactic acid bacteria was used as a placebo. The subjects ingested one bottle (100 mL) of the test food or placebo food per day for four weeks.

[0081] [Exam Schedule] Twenty subjects who were eligible to participate in this study were assigned under double-blind conditions to a test food intake group and a placebo intake group (10 subjects each), and after four weeks of continuous intake, the study was conducted as a randomized, double-blind, placebo-controlled study in which saccharin test values ​​were used as the primary endpoint and compared with the placebo group. During the study period, the intake of specified health foods, functional food products, and supplements was prohibited, but there were no other dietary restrictions.

[0082] [Saccharin test] Subjects were instructed to abstain from alcohol from the morning before the test until the end of the test, and to finish eating by 9 p.m. on the day before the test. After waking up on the day of each test, they refrained from drinking any fluids and fasted until the end of the test. At least one hour before the saccharin test, subjects were required to consume the test meal or placebo meal. On the day of the test, subjects were prohibited from performing any physical work or exercise until the end of the test. In the saccharin test, saccharin granules (approximately 1 mm in diameter), an artificial sweetener, were attached to the mucosa of one side of the subject's nasal septum, and the time it took for the sweetness to be recognized in the pharynx (saccharin time) was measured. During the measurement, eating, drinking, blowing one's nose, sniffing, sneezing, and coughing were prohibited, and the upper limit of measurement time was set at 60 minutes. If no sweetness was detected after 60 minutes, the measurement was terminated at that point. For subjects who did not detect sweetness, saccharin granules were placed directly on the tongue after the test to confirm whether they could detect the sweetness (to confirm whether the subject was able to detect sweetness).

[0083] [result] The results of the saccharin time measurements are shown in Figure 4. The test food intake group (11 / 19-B1 intake group) had a shorter saccharin time than the placebo group, indicating a tendency for faster mucociliary transport in the nasal cavity. As mentioned above, there is a correlation between mucociliary transport function in the nasal cavity and trachea, so it is thought that in humans, intake of 11 / 19-B1 lactic acid bacteria will also improve mucociliary transport in the trachea. [Industrial Applicability]

[0084] The mucociliary clearance enhancer of the present invention (food and beverage composition for enhancing mucociliary clearance) can be added to foods and beverages, and is widely used in the production of foods and beverages such as fermented milk, pudding, lactic acid bacteria drinks, ice cream, ice milk, lacto ice cream, and frozen desserts. [Accession number]

[0085] NITE BP-01694

Claims

1. A mucociliary clearance enhancer characterized by containing Lactococcus lactic acid bacteria as an active ingredient.

2. The mucociliary clearance enhancer according to claim 1, wherein the Lactococcus lactic acid bacteria are lactic acid bacteria belonging to Lactococcus lactis subsp. lactis.

3. 2. The mucociliary clearance enhancer according to claim 1, wherein the Lactococcus lactic acid bacterium is a lactic acid bacterium having an accession number of NITE BP-01694 at the National Patent Microorganisms Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE).

4. A food and drink composition for enhancing mucociliary clearance, characterized in that it contains Lactococcus lactic acid bacteria as an active ingredient.

5. The food and beverage composition for enhancing mucociliary clearance according to claim 4, wherein the Lactococcus lactic acid bacteria are lactic acid bacteria belonging to Lactococcus lactis subsp. lactis.

6. The food and beverage composition for enhancing mucociliary clearance according to claim 4, wherein the Lactococcus lactic acid bacteria is a lactic acid bacterium with an accession number of NITE BP-01694 at the National Patent Microorganisms Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE).

7. A food or drink for enhancing mucociliary clearance, comprising the food or drink composition for enhancing mucociliary clearance according to any one of claims 4 to 6.

8. The food or drink for enhancing mucociliary clearance according to claim 7, which is a dairy product.

9. The food or drink for enhancing mucociliary clearance according to claim 7, which is a fermented milk or lactic acid bacteria drink.

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