JP7372112B2 - Anti-type I allergy agent - Google Patents
Anti-type I allergy agent Download PDFInfo
- Publication number
- JP7372112B2 JP7372112B2 JP2019196388A JP2019196388A JP7372112B2 JP 7372112 B2 JP7372112 B2 JP 7372112B2 JP 2019196388 A JP2019196388 A JP 2019196388A JP 2019196388 A JP2019196388 A JP 2019196388A JP 7372112 B2 JP7372112 B2 JP 7372112B2
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- allergy
- lactic acid
- acid bacteria
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Description
特許法第３０条第２項適用 （その１） 発送日 ２０１８年１０月２９日 刊行物 第２１回日本補完代替医療学会学術集会 抄録集 （その２） 開催日 ２０１８年１１月１０日から２０１８年１１月１１日 集会名、開催場所 第２１回日本補完代替医療学会学術集会 日本獣医生命科学大学・日本医科大学武蔵境校舎（東京都武蔵野市境南町１－７－１） （その３） 開催日 ２０１９年４月１７日 集会名、開催場所 岡山市薬剤師会保険薬局部会研修会 岡山大学病院マスカットキューブ３Ｆ（岡山県岡山市北区鹿田町２－５－１） （その４） ウェブサイトの掲載日 ２０１９年９月１８日 ウェブサイトのアドレス ｈｔｔｐ：／／ｗｗｗ．ｃｃ．ｋｏｃｈｉ－ｕ．ａｃ．ｊｐ／▲～▼ｎｍｉｙａｋｅ／ａｍ６０／ｉｎｄｅｘ．ｈｔｍｌ ｈｔｔｐ：／／ｗｗｗ．ｃｃ．ｋｏｃｈｉ－ｕ．ａｃ．ｊｐ／▲～▼ｎｍｉｙａｋｅ／ａｍ６０／ａｍ６０＿ｐｒｏｇｒａｍ．ｐｄｆ （その５） 発行日 ２０１９年１０月１１日 刊行物 日本花粉学会第６０回大会講演要旨 （その６） 開催日 ２０１９年１０月１１日～２０１９年１０月１３日 集会名、開催場所 日本花粉学会第６０回大会 高知大学（朝倉キャンパス）（高知市曙町２－５－１） （その７） ウェブサイトの掲載日 ２０１９年１０月１５日 ウェブサイトのアドレス ｈｔｔｐｓ：／／ｗｗｗ．ｏｔｓｕｋａ．ｃｏ．ｊｐ／ｃｏｍｐａｎｙ／ｎｅｗｓｒｅｌｅａｓｅｓ／２０１９／２０１９１０１５＿１．ｈｔｍｌApplication of Article 30, Paragraph 2 of the Patent Act (Part 1) Shipping date: October 29, 2018 Publication Abstracts of the 21st Annual Meeting of the Japanese Society for Complementary and Alternative Medicine (Part 2) Date of event: From November 10, 2018 to 2018 November 11th Meeting name, venue 21st Japanese Society of Complementary and Alternative Medicine Academic Meeting Nippon Veterinary and Life Science University/Nippon Medical University Musashisakai Campus (1-7-1 Sakainancho, Musashino City, Tokyo) (Part 3) Date 2019 April 17, 2019 Meeting name and location Okayama City Pharmacists Association Health Insurance Pharmacy Section Training Session Okayama University Hospital Muscat Cube 3F (2-5-1 Shikata-cho, Kita-ku, Okayama City, Okayama Prefecture) (Part 4) Website publication date 2019 September 18, 2018 Website address http://www. cc. kochi-u. ac. jp/▲～▼nmiyake/am60/index. html http://www. cc. kochi-u. ac. jp/▲～▼nmiyake/am60/am60_program. pdf (Part 5) Publication date October 11, 2019 Publication Abstracts of the 60th Conference of the Japanese Pollen Society (Part 6) Event date October 11, 2019 to October 13, 2019 Meeting name and venue Japanese pollen 60th Conference of the Society Kochi University (Asakura Campus) (2-5-1 Akebono-cho, Kochi City) (Part 7) Website publication date October 15, 2019 Website address https://www. otsuka. co. jp/company/newsreleases/2019/20191015_1. html
本発明は、抗Ｉ型アレルギー剤、花粉症軽減剤、アレルゲンによる不快感の軽減剤、及びＩｇＥ低減剤に関する。 TECHNICAL FIELD The present invention relates to an anti-type I allergy agent, a hay fever alleviating agent, an allergen-induced discomfort alleviating agent, and an IgE reducing agent.
近年、日本の多くの人々が、スギ(Cryptomeria japonica)花粉によって引き起こされるアレルギー反応に苦しんでおり、その患者の数は、人口の約３分の１を占め、多くの対策が講じられているが増加している。したがって、花粉症は国民病であり、スギ花粉の季節には、毎日、スギ花粉の飛散予報がマスコミにより発表されている。 In recent years, many people in Japan have been suffering from allergic reactions caused by cedar (Cryptomeria japonica) pollen, accounting for about one-third of the population, and many countermeasures have been taken. It has increased. Therefore, hay fever is a national disease, and during the cedar pollen season, the mass media announces cedar pollen scattering forecasts every day.
くしゃみ、鼻水、鼻づまり及び目のかゆみのようなアレルギー反応は、スギ花粉曝露の最初の段階では現れないが、花粉曝露が反応に必要なしきい値を越えると発症する。したがって、アレルギー反応を最小限にするためには曝露を防ぐことが重要であり、完全にスギ花粉を防ぐことができなくても、多くの人々がスギ花粉の季節中にはマスクを着けている。 Allergic reactions such as sneezing, runny nose, stuffy nose, and itchy eyes do not appear during the initial stages of cedar pollen exposure, but develop when pollen exposure exceeds the threshold required for a reaction. Therefore, prevention of exposure is important to minimize allergic reactions, and even if it is not possible to completely prevent cedar pollen, many people wear masks during cedar pollen season. .
抗ヒスタミン剤又はステロイド点鼻薬のような抗アレルギー剤は、花粉曝露後の症状を緩和するために使用されるが、それらの効果は一時的であったり、眠気のような副作用を有している。舌下免疫療法は、日本においてスギ花粉又はダニアレルギーのための治療法として最近承認された。しかしながら、毎日、長期間の摂取が必要であり、口腔そう痒症、口腔浮腫、咽頭刺激感及びくしゃみのような局所的副作用が生じることが知られている。したがって、新しい治療方法の開発が求められている。 Antiallergic agents such as antihistamines or steroid nasal sprays are used to alleviate symptoms after pollen exposure, but their effects are temporary or have side effects such as drowsiness. Sublingual immunotherapy was recently approved as a treatment for cedar pollen or dust mite allergies in Japan. However, it requires daily, long-term ingestion and is known to cause local side effects such as oral itching, oral edema, pharyngeal irritation, and sneezing. Therefore, the development of new treatment methods is required.
アレルギー反応の重症度は幅広く、個人のアレルギー反応のタイプ及び重症度に応じて適切な治療が行われるべきである。例えば、軽度の反応を示す人では、アレルギー症状を減少させるだけでなく、ＱＯＬを改善する効果を有する抗アレルギー食品の摂取により、アレルギーをコントロールすることもできる。 Allergic reactions can vary in severity, and appropriate treatment should depend on the type and severity of the individual's allergic reaction. For example, people with mild reactions can control their allergies by ingesting anti-allergic foods that not only reduce allergic symptoms but also improve QOL.
乳酸菌の一種であるラクトバチルスＯＮＲＩＣｂ０２４０（ＦＥＲＭ ＢＰ－１００６５）株は、抗鳥インフルエンザ抗体産生促進作用（特許文献１）、肺炎予防、風邪予防、生活の質（Ｑｕａｌｉｔｙ ｏｆ Ｌｉｆｅ；ＱＯＬ）の改善作用（特許文献２）を有することが知られている。 Lactobacillus ONRIC b0240 (FERM BP-10065) strain, a type of lactic acid bacteria, has the effect of promoting the production of anti-avian influenza antibodies (Patent Document 1), preventing pneumonia, preventing colds, and improving quality of life (QOL). Patent Document 2).
本発明は、新たな抗Ｉ型アレルギー剤、花粉症軽減剤、アレルゲンによる不快感の軽減剤、及びＩｇＥ低減剤を提供することを目的とする。 An object of the present invention is to provide a new anti-type I allergy agent, a hay fever alleviating agent, an allergen-induced discomfort alleviating agent, and an IgE reducing agent.
本発明者らは、上記目的を達成すべく鋭意研究を重ねた結果、アレルギー性鼻炎モデルマウスを用いた試験とヒトの花粉に対するアレルギー反応についての試験により、ラクトバチルスＯＮＲＩＣｂ０２４０（ＦＥＲＭ ＢＰ－１００６５）がＩ型アレルギーに対する抗アレルギー作用を有することを見出した。 As a result of extensive research to achieve the above objective, the present inventors found that Lactobacillus ONRICb0240 (FERM BP-10065) It was found that it has an anti-allergic effect against type I allergy.
本発明は、これら知見に基づき、更に検討を重ねて完成されたものであり、次の抗Ｉ型アレルギー剤、花粉症軽減剤、アレルゲンによる不快感の軽減剤、及びＩｇＥ低減剤を提供するものである。 The present invention was completed based on these findings and further studies, and provides the following anti-type I allergy agent, hay fever alleviation agent, allergen-induced discomfort alleviation agent, and IgE reducing agent. It is.
（１）抗Ｉ型アレルギー剤
項１－１．ラクトバチルス・ペントーサスに属する乳酸菌を含有する、抗Ｉ型アレルギー剤。
項１－２．前記Ｉ型アレルギーが、アトピー性皮膚炎、アレルギー性鼻炎、花粉症、アレルギー性皮膚炎、気管支喘息、蕁麻疹、アレルギー性結膜炎、又はアナフィラキシーショックである、項１－１に記載の抗Ｉ型アレルギー剤。
項１－３．前記ラクトバチルス・ペントーサスに属する乳酸菌が、ラクトバチルスＯＮＲＩＣｂ０２４０（ＦＥＲＭ ＢＰ－１００６５）又はその変異株である、項１－１又は１－２に記載の抗Ｉ型アレルギー剤。
項１－４．飲食品又は医薬品である、項１－１～１－３のいずれか一項に記載の抗Ｉ型アレルギー剤。
(1) Anti-type I allergy agent Item 1-1. An anti-type I allergy agent containing lactic acid bacteria belonging to Lactobacillus pentosus.
Section 1-2. The anti-type I allergy according to item 1-1, wherein the type I allergy is atopic dermatitis, allergic rhinitis, hay fever, allergic dermatitis, bronchial asthma, urticaria, allergic conjunctivitis, or anaphylactic shock. agent.
Section 1-3. Item 1-1 or 1-2, wherein the lactic acid bacterium belonging to Lactobacillus pentosus is Lactobacillus ONRICb0240 (FERM BP-10065) or a mutant strain thereof.
Section 1-4. The anti-type I allergy agent according to any one of Items 1-1 to 1-3, which is a food or drink or a pharmaceutical.
（２）花粉症軽減剤
項２－１．ラクトバチルス・ペントーサスに属する乳酸菌を含有する、花粉症軽減剤。
項２－２．前記ラクトバチルス・ペントーサスに属する乳酸菌が、ラクトバチルスＯＮＲＩＣｂ０２４０（ＦＥＲＭ ＢＰ－１００６５）又はその変異株である、項２－１に記載の花粉症軽減剤。
項２－３．飲食品又は医薬品である、項２－１又は２－２に記載の花粉症軽減剤。
(2) Hay fever alleviation agent Section 2-1. A hay fever alleviation agent containing lactic acid bacteria belonging to Lactobacillus pentosus.
Section 2-2. Item 2-1, wherein the lactic acid bacteria belonging to Lactobacillus pentosus is Lactobacillus ONRICb0240 (FERM BP-10065) or a mutant strain thereof.
Section 2-3. The hay fever alleviation agent according to item 2-1 or 2-2, which is a food or drink or a medicine.
（３）アレルゲンによる不快感の軽減剤
項３－１．ラクトバチルス・ペントーサスに属する乳酸菌を含有する、アレルゲンによる不快感の軽減剤。
項３－２．前記アレルゲンが花粉である、項３－１に記載のアレルゲンによる不快感の軽減剤。
項３－３．前記ラクトバチルス・ペントーサスに属する乳酸菌が、ラクトバチルスＯＮＲＩＣｂ０２４０（ＦＥＲＭ ＢＰ－１００６５）又はその変異株である、項３－１又は３－２に記載のアレルゲンによる不快感の軽減剤。
項３－４．飲食品又は医薬品である、項３－１～３－３のいずれか一項に記載のアレルゲンによる不快感の軽減剤。
(3) Agent for alleviating discomfort caused by allergen Item 3-1. An agent that reduces discomfort caused by allergens, containing lactic acid bacteria belonging to Lactobacillus pentosus.
Section 3-2. Item 3-1. The agent for alleviating discomfort caused by an allergen according to item 3-1, wherein the allergen is pollen.
Section 3-3. Item 3-1 or 3-2, wherein the lactic acid bacterium belonging to Lactobacillus pentosus is Lactobacillus ONRICb0240 (FERM BP-10065) or a mutant strain thereof.
Section 3-4. The agent for alleviating discomfort caused by an allergen according to any one of Items 3-1 to 3-3, which is a food or drink or a pharmaceutical.
（４）ＩｇＥ低減剤
項４－１．ラクトバチルス・ペントーサスに属する乳酸菌を含有する、ＩｇＥ低減剤。
項４－２．前記ラクトバチルス・ペントーサスに属する乳酸菌が、ラクトバチルスＯＮＲＩＣｂ０２４０（ＦＥＲＭ ＢＰ－１００６５）又はその変異株である、項４－１に記載のＩｇＥ低減剤。
項４－３．飲食品又は医薬品である、項４－１又は４－２に記載のＩｇＥ低減剤。
(4) IgE reducing agent Item 4-1. An IgE reducing agent containing lactic acid bacteria belonging to Lactobacillus pentosus.
Section 4-2. Item 4-1. The IgE reducing agent according to item 4-1, wherein the lactic acid bacterium belonging to Lactobacillus pentosus is Lactobacillus ONRICb0240 (FERM BP-10065) or a mutant strain thereof.
Section 4-3. The IgE reducing agent according to item 4-1 or 4-2, which is a food or drink or a pharmaceutical.
ラクトバチルス・ペントーサスに属する乳酸菌は優れたＩ型アレルギーに対する抗アレルギー作用を有するので、抗Ｉ型アレルギー剤、花粉症軽減剤、及びアレルゲンによる不快感の軽減剤の有効成分として有用である。また、より詳細には、ラクトバチルス・ペントーサスに属する乳酸菌は優れたＩｇＥ低減作用を有するので、ＩｇＥ低減剤の有効成分として有用である。そのため、本発明によれば、ラクトバチルス・ペントーサスに属する乳酸菌を有効成分とする、抗Ｉ型アレルギー剤、花粉症軽減剤、アレルゲンによる不快感の軽減剤、及びＩｇＥ低減剤を提供することができる。 Lactic acid bacteria belonging to Lactobacillus pentosus have excellent anti-allergic activity against type I allergies, and are therefore useful as active ingredients in anti-type I allergy agents, hay fever alleviation agents, and agents for allergen-induced discomfort. More specifically, lactic acid bacteria belonging to Lactobacillus pentosus have an excellent IgE-reducing effect and are therefore useful as an active ingredient of an IgE-reducing agent. Therefore, according to the present invention, it is possible to provide an anti-type I allergy agent, a hay fever alleviation agent, an allergen-induced discomfort alleviation agent, and an IgE reducing agent that contain lactic acid bacteria belonging to Lactobacillus pentosus as an active ingredient. .
以下、本発明の実施の形態について説明する。 Embodiments of the present invention will be described below.
本発明の抗Ｉ型アレルギー剤、花粉症軽減剤、アレルゲンによる不快感の軽減剤、及びＩｇＥ低減剤は、ラクトバチルス・ペントーサスに属する乳酸菌を有効成分として含有することを特徴とする。 The anti-type I allergy agent, hay fever alleviation agent, allergen-induced discomfort alleviation agent, and IgE reducing agent of the present invention are characterized by containing lactic acid bacteria belonging to Lactobacillus pentosus as an active ingredient.
本発明におけるＩ型アレルギーとしては、特に制限されないが、アトピー性皮膚炎、アレルギー性鼻炎、花粉症、アレルギー性皮膚炎、気管支喘息、蕁麻疹、アレルギー性結膜炎、アナフィラキシーショックなどが挙げられる。 Type I allergies in the present invention include, but are not particularly limited to, atopic dermatitis, allergic rhinitis, hay fever, allergic dermatitis, bronchial asthma, urticaria, allergic conjunctivitis, and anaphylactic shock.
本発明におけるアレルゲンとしては、特に制限されないが、ハウスダスト、ほこり、花粉、ダニ、カビ、細菌及び動物のフケなどが挙げられ、特に花粉である。花粉の種類としては、特に制限されないが、スギ、ヒノキ、ハンノキ、シラカンバ、サワラ、ブタクサ、ヨモギ、カナムグラ、イネ科植物などの花粉が挙げられ、特にスギ花粉である。 Examples of allergens in the present invention include, but are not limited to, house dust, dust, pollen, mites, mold, bacteria, and animal dander, and pollen is particularly preferred. The types of pollen include, but are not particularly limited to, pollens of cedar, cypress, alder, birch, Japanese Spanish mackerel, ragweed, mugwort, Japanese grass, grasses, and cedar pollen in particular.
本発明のアレルゲンによる不快感の軽減剤における不快感としては、例えば、目、鼻のアレルギー反応及びＱＯＬ低下を総括した不快感であり、具体的には、鼻水、くしゃみ、鼻づまり、鼻のかゆみ、目のかゆみ、流涙などの症状やアレルゲン曝露によるＱＯＬ低下を含む総括的な不快感が挙げられる。 The discomfort in the agent for allergen-induced discomfort of the present invention is, for example, the discomfort that is a generalization of allergic reactions in the eyes and nose and decreased QOL, and specifically, the discomfort includes a runny nose, sneezing, stuffy nose, and itchy nose. , symptoms such as itching and tearing of the eyes, and general discomfort including a decline in QOL due to allergen exposure.
本発明において使用されるラクトバチルス・ペントーサス(Lactobacillus pentosus)としては、例えば、ラクトバチルス・ペントーサスＳ－ＰＴ８４、ラクトバチルスＯＮＲＩＣｂ０２４０株などが挙げられ、好ましくはラクトバチルスＯＮＲＩＣｂ０２４０株である。ラクトバチルス・ペントーサスに属する乳酸菌は、１種又は２種以上を任意に組み合わせて使用することができる。 Examples of Lactobacillus pentosus used in the present invention include Lactobacillus pentosus S-PT84 and Lactobacillus ONRICb0240 strain, and Lactobacillus ONRICb0240 strain is preferred. Lactic acid bacteria belonging to Lactobacillus pentosus can be used alone or in any combination of two or more.
ラクトバチルスＯＮＲＩＣｂ０２４０（以下、Ｂ２４０と表記することもある）株は、天然物から単離された乳酸菌であり、平成１５年８月６日に、日本国茨城県つくば市東１丁目１番地１ 中央第６に住所を有する独立行政法人 産業技術総合研究所 特許生物寄託センター（ＩＰＯＤ）に寄託番号ＦＥＲＭ Ｐ－１９４７０として寄託され、現在、国際寄託に移管されており、その国際寄託番号はＦＥＲＭ ＢＰ－１００６５である。なお、独立行政法人産業技術総合研究所特許生物寄託センターは、２０１２年４月に独立行政法人製品評価技術基盤機構（ＮＩＴＥ）特許微生物寄託センターと統合され、現在、独立行政法人製品評価技術基盤機構バイオテクノロジーセンター特許生物寄託センター（ＮＩＴＥ－ＩＰＯＤ）（〒２９２－０８１８ 日本国千葉県木更津市かずさ鎌足２－５－８ １２０号室）にてその微生物寄託業務は承継されている。ラクトバチルスＯＮＲＩＣｂ０２４０株の菌学的性質については、既知である。なお、本発明で使用されるラクトバチルスＯＮＲＩＣｂ０２４０株は、国際寄託の時点では、ラクトバチルス・プランタラム(Lactobacillus plantarum)に属すると分類されていたが、その後の分類基準の変更(Francois Bringle et al., International Journal of Systematic and Evolutionary Microbiology, Vol. 55, 2005, p.1629-1634)に伴い、本株は、ラクトバチルス・ペントーサス(Lactobacillus pentosus)に分類されることになった。 Lactobacillus ONRIC b0240 (hereinafter sometimes referred to as B240) is a lactic acid bacterium isolated from natural products, and was first published on August 6, 2003 at Chuo Station, Higashi 1-1-1, Tsukuba City, Ibaraki Prefecture, Japan. It was deposited with the International Patent Organism Depositary (IPOD) of the National Institute of Advanced Industrial Science and Technology (AIST), an independent administrative agency with an address at 6, under the deposit number FERM P-19470, and has now been transferred to the International Deposit, and its international deposit number is FERM BP-10065. It is. The National Institute of Advanced Industrial Science and Technology (AIST) Patent Organism Depositary was merged with the National Institute of Technology and Product Evaluation (NITE) Patent Microorganism Depositary in April 2012, and is currently part of the National Institute of Technology and Product Evaluation (NITE). The microbial deposit service has been taken over by the Biotechnology Center Patent Organism Depositary (NITE-IPOD) (Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan). The mycological properties of Lactobacillus ONRICb0240 strain are known. The Lactobacillus ONRICb0240 strain used in the present invention was classified as belonging to Lactobacillus plantarum at the time of international deposit, but the classification criteria were subsequently changed (Francois Bringle et al. , International Journal of Systematic and Evolutionary Microbiology, Vol. 55, 2005, p.1629-1634), this strain was classified as Lactobacillus pentosus.
ラクトバチルスＯＮＲＩＣｂ０２４０株の変異株としては、ラクトバチルスＯＮＲＩＣｂ０２４０株から得られた任意の株であって、ラクトバチルスＯＮＲＩＣｂ０２４０株と同様に抗アレルギー作用を有するものである限り特に制限されない。このような変異株は、例えば、自然突然変異、化学的若しくは物理的変異原による誘発変異によって人工的に突然変異の頻度を高める方法、又は遺伝子組換え技術により得ることができる。変異原の例としては、例えば、エチルメタンスルホネート、亜硝酸、Ｎ－メチル－Ｎ’－ニトロ－Ｎ－ニトロソグアニジン、ＵＶ照射、重イオンビーム照射、Ｘ線照射、ガンマ線照射などが挙げられる。 The mutant strain of Lactobacillus ONRICb0240 is not particularly limited as long as it is any strain obtained from Lactobacillus ONRICb0240 and has an antiallergic effect like Lactobacillus ONRICb0240. Such mutant strains can be obtained, for example, by natural mutation, by a method of artificially increasing the mutation frequency by chemical or physical mutagen-induced mutation, or by genetic recombination technology. Examples of mutagens include, for example, ethyl methanesulfonate, nitrous acid, N-methyl-N'-nitro-N-nitrosoguanidine, UV irradiation, heavy ion beam irradiation, X-ray irradiation, gamma ray irradiation, and the like.
本発明の抗Ｉ型アレルギー剤、花粉症軽減剤、アレルゲンによる不快感の軽減剤、及びＩｇＥ低減剤（以下、抗Ｉ型アレルギー剤等と表記することもある）に含有されるラクトバチルス・ペントーサスに属する乳酸菌は、生菌の状態であってもよく、死菌の状態であってもよく、菌体処理物であってもよく、また、これらが混合された状態であってもよい。ここで、生菌とは、生きた状態の乳酸菌であり、乳酸菌の培養液、当該培養液の懸濁物、粗精製物、精製物や、また、これらの生きた乳酸菌を凍結乾燥やスプレードライ等により乾燥させた菌体粉末なども含まれ、生きた状態である限り制限されない。また、死菌とは、生きた状態の乳酸菌を加熱処理、放射線処理等の物理的処理又は化学的処理に供して殺菌された状態の乳酸菌であり、殺菌された状態の乳酸菌が凍結乾燥やスプレードライ等により乾燥された菌体粉末なども含まれ、死菌である限り制限されない。また、菌体処理物とは、乳酸菌をホモジナイズ処理、酵素処理、超音波処理等で破壊した菌体破砕物であり、当該菌体破砕物を凍結乾燥やスプレードライ等により乾燥させた菌体破砕物の粉末も含まれる。本発明に含有されているラクトバチルス・ペントーサスに属する乳酸菌は、好ましくは生菌、死菌、菌体処理物又はこれらの混合物の状態であり、より好ましくは死菌又は生菌と死菌の混合物の状態であり、更に好ましくは死菌の状態ある。 Lactobacillus pentosus contained in the anti-type I allergy agent, hay fever allergy reducer, allergen-induced discomfort reducer, and IgE reducer (hereinafter sometimes referred to as anti-type I allergy agent, etc.) of the present invention. The lactic acid bacteria belonging to this group may be in a live state, a dead state, a processed product of bacterial cells, or a mixture thereof. Here, the term "live bacteria" refers to lactic acid bacteria in a living state, such as a culture solution of lactic acid bacteria, a suspension of the culture solution, a crude product, a purified product, and a method to freeze-dry or spray-dry these live lactic acid bacteria. It also includes dried bacterial powder, etc., and is not limited as long as it is in a living state. In addition, dead bacteria refers to lactic acid bacteria that have been sterilized by subjecting living lactic acid bacteria to physical treatment such as heat treatment or radiation treatment, or chemical treatment, and sterilized lactic acid bacteria can be sterilized by freeze-drying or spraying. It also includes dried bacterial powder, etc., and is not limited as long as the bacteria are dead. In addition, a bacterial cell treatment product is a bacterial cell fragment obtained by destroying lactic acid bacteria by homogenization treatment, enzyme treatment, ultrasonic treatment, etc., and a bacterial cell fragment obtained by drying the bacterial cell fragment by freeze-drying, spray drying, etc. It also includes powdered substances. The lactic acid bacteria belonging to Lactobacillus pentosus contained in the present invention are preferably in the form of live bacteria, dead bacteria, treated bacteria, or a mixture thereof, and more preferably dead bacteria or a mixture of live bacteria and dead bacteria. More preferably, the bacteria are dead.
本発明の抗Ｉ型アレルギー剤等に使用されるラクトバチルス・ペントーサスに属する乳酸菌は、当該乳酸菌株の生育に適した培地で培養することによって増殖させることができる。培養方法は制限されないが、例えば、ＭＲＳ培地、ＬＢＳ培地、Ｒｏｇｏｓａ培地等の培地中で３０℃で１６時間程度培養することによって増殖させることができる。また、菌体は、培養後に、例えば、培養物（培養液）を遠心分離（例えば、３，０００ｒｐｍ、４℃、１０分間）して集菌することができる。また、本発明で使用されるラクトバチルス・ペントーサスに属する乳酸菌は、乳、野菜類、果物類、豆乳等などの素材等の存在下で培養（発酵）してもよい。前述同様、菌体は、培養後に遠心分離を行うことにより集菌することができる。本発明においては、このようにして得られる培養物（発酵物）や集菌された菌体、当該培養物又は菌体の懸濁物や濃縮物、また、このようにして得られた培養物、菌体、懸濁物、濃縮物を凍結乾燥やスプレードライ等により乾燥させて粉末化したものなども用いることもできる。これらの調製は、当業界で公知の方法に従って行うことができる。また、培養（発酵）がより効率よく行われる点から、発酵前の乳、野菜類、果物類、豆乳などの素材は、液状など一定以上の流動性を有することが好ましい。 The lactic acid bacteria belonging to Lactobacillus pentosus used in the anti-type I allergy agent of the present invention can be grown by culturing in a medium suitable for the growth of the lactic acid bacteria strain. Although the culture method is not limited, for example, the cells can be grown by culturing at 30° C. for about 16 hours in a medium such as MRS medium, LBS medium, Rogosa medium, or the like. Furthermore, after culturing, the bacterial cells can be collected by, for example, centrifuging the culture (culture solution) (for example, at 3,000 rpm, 4° C., for 10 minutes). Furthermore, the lactic acid bacteria belonging to Lactobacillus pentosus used in the present invention may be cultured (fermented) in the presence of materials such as milk, vegetables, fruits, soy milk, and the like. As described above, the bacterial cells can be collected by centrifugation after culturing. In the present invention, the culture (fermented product) obtained in this way, the collected bacterial cells, the suspension or concentrate of the culture or bacterial cells, and the culture obtained in this way It is also possible to use powdered products obtained by drying bacterial cells, suspensions, or concentrates by freeze-drying, spray drying, or the like. These preparations can be performed according to methods known in the art. Further, in order to perform culture (fermentation) more efficiently, it is preferable that the raw material such as milk, vegetables, fruits, soy milk, etc. before fermentation has a fluidity above a certain level, such as in a liquid state.
本発明の抗Ｉ型アレルギー剤等は、前述するように、ラクトバチルス・ペントーサスに属する乳酸菌を有効成分として含有すればよい。したがって、例えば、前記培養物をそのまま、あるいは均質化等の処理を施して、抗Ｉ型アレルギー剤等として使用してもよいし、前記調製したものを抗Ｉ型アレルギー剤等として使用してもよい。 As mentioned above, the anti-type I allergy agent of the present invention may contain lactic acid bacteria belonging to Lactobacillus pentosus as an active ingredient. Therefore, for example, the culture may be used as it is or after being subjected to a treatment such as homogenization as an anti-type I allergy agent, or the prepared product may be used as an anti-type I allergy agent, etc. good.
本発明の抗Ｉ型アレルギー剤等に、生きた状態のラクトバチルス・ペントーサスに属する乳酸菌が含有されている場合、生きた状態を一層良好に維持させる点から、必要に応じて更に、ラクトバチルス・ペントーサスに属する乳酸菌の生育に適した栄養成分を抗Ｉ型アレルギー剤等に含有させることが望ましい。当該栄養成分としては、例えば、グルコース、澱粉、蔗糖、乳糖、デキストリン、ソルビトール、フラクトース等の炭素源；酵母エキス、ペプトン等の窒素源；ビタミン類；ミネラル類；微量金属元素；その他の栄養成分等の各成分を挙げることができる。ビタミン類としては、具体的には、ビタミンＢ、ビタミンＤ、ビタミンＣ、ビタミンＥ、ビタミンＫ等を例示できる。微量金属元素としては、具体的には、亜鉛、セレン等を例示できる。その他の栄養成分としては、具体的には、乳果オリゴ糖、大豆オリゴ糖、ラクチュロース、ラクチトール、フラクトオリゴ糖、ガラクトオリゴ糖等の各種オリゴ糖を例示できる。 When the anti-type I allergy agent of the present invention contains lactic acid bacteria belonging to Lactobacillus pentosus in a living state, in order to better maintain the living state, Lactobacillus pentosus may be added as necessary. It is desirable that the anti-type I allergy agent etc. contain nutritional components suitable for the growth of lactic acid bacteria belonging to Pentosus. The nutritional components include, for example, carbon sources such as glucose, starch, sucrose, lactose, dextrin, sorbitol, and fructose; nitrogen sources such as yeast extract and peptone; vitamins; minerals; trace metal elements; other nutritional components, etc. Each component can be mentioned. Specific examples of the vitamins include vitamin B, vitamin D, vitamin C, vitamin E, and vitamin K. Specific examples of trace metal elements include zinc, selenium, and the like. Specific examples of other nutritional components include various oligosaccharides such as milk oligosaccharide, soybean oligosaccharide, lactulose, lactitol, fructooligosaccharide, and galactooligosaccharide.
また、本発明の抗Ｉ型アレルギー剤等には、更に必要に応じて任意の成分を含有させることができ、当該任意の成分として、例えば、可食性又は薬学的に許容される担体や添加物等を含有させることができる。可食性又は薬学的に許容される担体や添加物としては、水性媒体、賦形剤、結合剤、崩壊剤、滑沢剤、増粘剤、界面活性剤、浸透圧調節剤、付湿剤、ｐＨ調整剤、甘味料、香料、色素等が例示される。これらは当業者により公知であり、適宜選択して使用される。具体的には、これらの一例として、水、生理食塩水、果汁等の水性媒体；乳糖、白糖、塩化ナトリウム、ブドウ糖、尿素、デンプン、炭酸カルシウム、カオリン、結晶セルロース、ケイ酸、リン酸カリウム、コーンスターチ、デキストリン等の賦形剤；水、エタノール、プロパノール、単シロップ、ブドウ糖液、デンプン液、ゼラチン溶液、カルボキシメチルセルロース、ヒドロキシプロピルセルロース、メチルセルロース、ポリビニルピロリドン等の結合剤；カルボキシメチルセルロースナトリウム、カルボキシメチルセルロースカルシウム、低置換度ヒドロキシプロピルセルロース、乾燥デンプン、アルギン酸ナトリウム、カンテン末、ラミナラン末、炭酸水素ナトリウム、炭酸カルシウム等の崩壊剤；精製タルク、ステアリン酸塩、ホウ酸末、ポリエチレングリコール等の滑沢剤；ゼラチン、アラビアガム、デキストリン、メチルチセルロース、ポリビニルピロリドン、ポリビニルアルコール、ヒドロキシプロピルセルロース、キサンタンガム、ペクチン、トラガントガム、カゼイン、アルギン酸等の増粘剤；リオキシエチレンソルビタン脂肪酸エステル類、ラウリル硫酸ナトリウム、ステアリン酸モノグリセリド等の界面活性剤；ステビア、サッカリン、アセスルファムＫ、アスパラテーム、スクラロース等の甘味料等が挙げられる。 In addition, the anti-type I allergy agent of the present invention can further contain arbitrary components as necessary, such as edible or pharmaceutically acceptable carriers and additives. etc. can be contained. Edible or pharmaceutically acceptable carriers and additives include aqueous media, excipients, binders, disintegrants, lubricants, thickeners, surfactants, osmotic pressure regulators, wetting agents, Examples include pH adjusters, sweeteners, flavors, and pigments. These are known to those skilled in the art, and are appropriately selected and used. Specifically, examples of these include aqueous media such as water, physiological saline, and fruit juice; lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid, potassium phosphate, Excipients such as corn starch and dextrin; binders such as water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, and polyvinylpyrrolidone; carboxymethylcellulose sodium, carboxymethylcellulose calcium , low-substituted hydroxypropyl cellulose, dry starch, sodium alginate, agar powder, laminaran powder, sodium bicarbonate, calcium carbonate, and other disintegrants; purified talc, stearate, boric acid powder, and polyethylene glycol and other lubricants; Thickeners such as gelatin, gum arabic, dextrin, methylthicellulose, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropylcellulose, xanthan gum, pectin, gum tragacanth, casein, alginic acid; lyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid Examples include surfactants such as monoglycerides; sweeteners such as stevia, saccharin, acesulfame K, aspartame, and sucralose.
必要に応じて使用される成分は当業者であれは適宜選択可能であり、また、当該成分の配合量は、目的とする形態や嗜好等に適合するよう、本発明の効果を妨げない範囲で、適宜調整すればよい。 Those skilled in the art can appropriately select the ingredients to be used as necessary, and the amount of the ingredients to be blended can be adjusted to suit the intended form and taste, within a range that does not impede the effects of the present invention. , may be adjusted as appropriate.
本発明の抗Ｉ型アレルギー剤等の形態は特に制限されないが、例えば、散剤、顆粒剤、錠剤、丸剤、トローチ等の固形形態；ゼリー、ムース、ヨーグルト、プディング、クリームのような半固形形態、液剤、懸濁剤、乳剤、シロップ剤等の液状形態が挙げられる。また、これらの形態は、マイクロカプセル、ソフトカプセル、ハードカプセル等に充填されて、カプセル剤の形態とすることもできる。また、本発明の抗Ｉ型アレルギー剤等は、発泡製剤の形態とすることもできる。これらの形態の製造方法は、当業界で公知の方法に従って行うことができる。 The form of the anti-type I allergy agent of the present invention is not particularly limited, and examples include solid forms such as powders, granules, tablets, pills, and troches; semi-solid forms such as jelly, mousse, yogurt, pudding, and cream. , liquid forms such as solutions, suspensions, emulsions, and syrups. Moreover, these forms can also be filled into microcapsules, soft capsules, hard capsules, etc. to form capsules. Moreover, the anti-type I allergy agent of the present invention can also be in the form of a foaming preparation. These forms can be manufactured according to methods known in the art.
本発明の抗Ｉ型アレルギー剤等におけるラクトバチルス・ペントーサスに属する乳酸菌の含有量は、１日当たりの投与量、投与形態、投与回数、使用目的等に応じて適宜設定される。本発明の効果が奏されることを限度として制限されないが、例えば、本発明の抗Ｉ型アレルギー剤等におけるラクトバチルス・ペントーサスに属する乳酸菌の含有量は、ラクトバチルス・ペントーサスに属する乳酸菌が総菌数（すなわち、生菌、死菌、菌体処理物の合計数）として１０４ｃｅｌｌｓ／ｍｇ以上、好ましくは１０５～１０１２ｃｅｌｌｓ／ｍｇ、より好ましくは１０６～１０１１ｃｅｌｌｓ／ｍｇである。 The content of lactic acid bacteria belonging to Lactobacillus pentosus in the anti-type I allergy agent of the present invention is appropriately set depending on the daily dosage, dosage form, frequency of administration, purpose of use, etc. For example, the content of lactic acid bacteria belonging to Lactobacillus pentosus in the anti-type I allergy agent of the present invention is not limited to the extent that the effects of the present invention are exhibited. The number (i.e., the total number of live bacteria, dead bacteria, and treated bacterial cells) is 10 4 cells/mg or more, preferably 10 5 to 10 12 cells/mg, and more preferably 10 6 to 10 11 cells/mg. .
また、本発明の抗Ｉ型アレルギー剤等におけるラクトバチルス・ペントーサスに属する乳酸菌の含有量は、本発明の効果が奏されることを限度として制限されないが、例えば、当該剤あたり、ラクトバチルス・ペントーサスに属する乳酸菌が総菌数して１０４ｃｅｌｌｓ以上、好ましくは１０６～１０１２ｃｅｌｌｓ、より好ましくは１０７～１０１２ｃｅｌｌｓ、更に好ましくは１０８～１０１２ｃｅｌｌｓ、効率良く所望の効果が得られる点から特に好ましくは総菌数として１０８～１０１１ｃｅｌｌｓ、一層好ましくは総菌数として１０９～１０１０ｃｅｌｌｓである。 In addition, the content of lactic acid bacteria belonging to Lactobacillus pentosus in the anti-type I allergy agent of the present invention is not limited as long as the effects of the present invention are achieved. The total number of lactic acid bacteria belonging to the group is 10 4 cells or more, preferably 10 6 to 10 12 cells, more preferably 10 7 to 10 12 cells, even more preferably 10 8 to 10 12 cells, and the desired effect can be efficiently obtained. In view of this, the total number of bacteria is particularly preferably 10 8 to 10 11 cells, and even more preferably 10 9 to 10 10 cells.
また、本発明の抗Ｉ型アレルギー剤等の投与量は、年齢、性別、症状等によって適宜調整されるが、成人１日当たりの投与量として、ラクトバチルス・ペントーサスに属する乳酸菌が総菌数として１０４ｃｅｌｌｓ以上、好ましくは１０６～１０１２ｃｅｌｌｓ、より好ましくは１０７～１０１２ｃｅｌｌｓ、更に好ましくは１０８～１０１２ｃｅｌｌｓ、効率良く所望の効果が得られる点から特に好ましくは１０８～１０１１ｃｅｌｌｓ、一層好ましくは１０９～１０１０ｃｅｌｌｓが例示される。これは、１日に１回又は数回に分けて投与することができる。投与方法は、本発明の効果を奏する限り制限されないが、経口投与が好ましい。 In addition, the dosage of the anti-type I allergy agent of the present invention is appropriately adjusted depending on age, sex, symptoms, etc., but the total number of lactic acid bacteria belonging to Lactobacillus pentosus is 10 per day for an adult. 4 cells or more, preferably 10 6 to 10 12 cells, more preferably 10 7 to 10 12 cells, still more preferably 10 8 to 10 12 cells, and particularly preferably 10 8 to 10 from the viewpoint of efficiently obtaining the desired effect. 11 cells, more preferably 10 9 to 10 10 cells. This can be administered once a day or in several divided doses. The administration method is not limited as long as it achieves the effects of the present invention, but oral administration is preferred.
なお、ラクトバチルス・ペントーサスに属する乳酸菌の総菌数は、公知の細菌数測定法や細菌数測定装置を使用して測定すればよい。例えば、公知の細菌数測定装置として顕微鏡、フローサイトメーター、また迅速検査装置バイオプローラ（登録商標）（パナソニックエコシステムズ社製）などが使用できる。後述する実施例においては、凍結乾燥原末を製造した段階でフローサイトメーターを使用して総菌数を測定した。 Note that the total number of lactic acid bacteria belonging to Lactobacillus pentosus may be measured using a known bacteria count measurement method or a bacteria count measurement device. For example, a microscope, a flow cytometer, a rapid testing device BioProra (registered trademark) (manufactured by Panasonic Ecosystems, Inc.), and the like can be used as known bacterial count measuring devices. In the Examples described below, the total number of bacteria was measured using a flow cytometer at the stage of manufacturing the freeze-dried bulk powder.
また、本発明における適用対象は、Ｉ型アレルギー症状を緩和することを目的とする動物であれば制限されない。例えば、当該動物としてヒトなどの哺乳動物が挙げられるが、このほか、哺乳動物以外のペットや家畜をはじめとする多様な動物が挙げられる。本発明は、適用対象の年齢や性別を問わない。 Moreover, the subject of the present invention is not limited as long as it is an animal whose purpose is to alleviate type I allergy symptoms. For example, such animals include mammals such as humans, but also various other animals such as pets and livestock other than mammals. The present invention is applicable to any age or gender.
本発明の抗Ｉ型アレルギー剤等は飲食品に応用できる。すなわち、本発明の抗Ｉ型アレルギー剤等は、そのまま飲食品として使用できる。また、飲食品への添加剤として使用できる。このような本発明の抗Ｉ型アレルギー剤等を含有する飲食品によれば、本発明の抗Ｉ型アレルギー剤等に起因する効果、すなわち、ラクトバチルス・ペントーサスに属する乳酸菌に起因する前記効果が得られる。また、本発明の抗Ｉ型アレルギー剤等は、そのまま医薬品として使用できる。また、本発明の抗Ｉ型アレルギー剤等は医薬品への添加剤として使用できる。このような本発明の抗Ｉ型アレルギー剤等を含有する医薬品によれば、本発明の抗Ｉ型アレルギー剤等に起因する効果、すなわち、ラクトバチルス・ペントーサスに属する乳酸菌に起因する前記効果が得られる。 The anti-type I allergy agent of the present invention can be applied to foods and drinks. That is, the anti-type I allergy agent of the present invention can be used as is as a food or drink. It can also be used as an additive to foods and drinks. According to the food/beverage products containing the anti-type I allergy agent of the present invention, the effect caused by the anti-type I allergy agent of the present invention, that is, the effect caused by lactic acid bacteria belonging to Lactobacillus pentosus, is suppressed. can get. Further, the anti-type I allergy agent of the present invention can be used as a pharmaceutical as it is. Furthermore, the anti-type I allergy agent of the present invention can be used as an additive to pharmaceuticals. According to such a pharmaceutical containing the anti-type I allergy agent of the present invention, the effect caused by the anti-type I allergy agent of the present invention, that is, the effect caused by lactic acid bacteria belonging to Lactobacillus pentosus can be obtained. It will be done.
本発明の抗Ｉ型アレルギー剤等がこのように飲食品又は医薬品に応用される場合、当該飲食品又は医薬品の種類は限定されず、ラクトバチルス・ペントーサスに属する乳酸菌を一成分として飲食品や医薬品に配合すればよく、また、必要に応じて任意の成分、例えば、可食性又は薬学的に許容される担体や添加物等を更に含有させることができる。可食性又は薬学的に許容される担体や添加物等としては、水性媒体、賦形剤、結合剤、崩壊剤、滑沢剤、増粘剤、界面活性剤、浸透圧調節剤、付湿剤、ｐＨ調整剤、甘味料、香料、色素等の前述のものが例示されるがこれらに限定されない。なお、これらの成分は当業者であれは適宜選択可能であり、また、当該成分の配合量は、目的とする形態や嗜好等に適合するよう、本発明の効果を妨げない範囲で、適宜調整すればよい。 When the anti-type I allergy agent of the present invention is applied to food/beverage products or medicines, the type of the food/beverage products or medicines is not limited. Furthermore, optional components such as edible or pharmaceutically acceptable carriers and additives may be further included as necessary. Edible or pharmaceutically acceptable carriers and additives include aqueous media, excipients, binders, disintegrants, lubricants, thickeners, surfactants, osmotic pressure regulators, and wetting agents. Examples include, but are not limited to, those mentioned above, such as pH adjusters, sweeteners, fragrances, and pigments. Those skilled in the art can appropriately select these ingredients, and the amount of the ingredients can be adjusted as appropriate to suit the intended form, taste, etc., to the extent that it does not impede the effects of the present invention. do it.
また、このように本発明の抗Ｉ型アレルギー剤等が飲食品又は医薬品に応用される場合の飲食品又は医薬品も本発明の効果を奏する限り制限されない。例えば、飲食品として、制限されないが、菓子類（ガム、キャンディー、クッキー、グミ、せんべい、ビスケット、ゼリー、ムース、クリームキャラメル、ラムネ菓子、可食性シート、可食性フィルム、トローチ等）、口中清涼剤（ガム、飴、グミ、可食性フィルム、トローチ等）、飲料（炭酸飲料、清涼飲料、乳飲料、アルコール飲料、果汁飲料、茶類、栄養飲料等）、粉末飲料（粉末ジュース、粉末スープ等）、乳製品（チーズ、ヨーグルト等）、パン、麺類、シリアル等が挙げられる。また、飲食品として、例えば、特定保健用食品、栄養補助食品、サプリメント、機能性表示食品、病者用食品等を挙げることができる。また、医薬品についても限定されないが、前述するような固形形態、半固形形態、液状形態の製剤、カプセル剤、発泡製剤等が挙げられる。これらの製造方法は、当業界で公知の方法に従って行うことができる。 Furthermore, in the case where the anti-type I allergy agent of the present invention is applied to food/beverage products or medicines, the food/beverage products or medicines are not limited as long as they exhibit the effects of the present invention. Examples of food and beverages include, but are not limited to, confectionery (gum, candies, cookies, gummies, rice crackers, biscuits, jellies, mousses, cream caramels, ramune sweets, edible sheets, edible films, troches, etc.), mouth fresheners, etc. (gum, candies, gummies, edible films, pastilles, etc.), beverages (carbonated drinks, soft drinks, milk drinks, alcoholic drinks, fruit juice drinks, teas, nutritional drinks, etc.), powdered drinks (powdered juices, powdered soups, etc.) , dairy products (cheese, yogurt, etc.), bread, noodles, cereals, etc. Furthermore, examples of the food and drink include foods for specified health uses, nutritional supplements, supplements, foods with functional claims, foods for the sick, and the like. Furthermore, pharmaceuticals include, but are not limited to, solid, semi-solid, and liquid preparations, capsules, foamed preparations, and the like as described above. These manufacturing methods can be performed according to methods known in the art.
本発明の抗Ｉ型アレルギー剤等を含有する飲食品又は医薬品におけるラクトバチルス・ペントーサスに属する乳酸菌の含有量は、１日当たりの投与量、投与形態、投与回数、使用目的等に応じて適宜設定される。本発明の効果が奏されることを限度として制限されないが、例えば、当該飲食品又は医薬品あたり、ラクトバチルス・ペントーサスに属する乳酸菌が総菌数として１０４ｃｅｌｌｓ／ｍｇ以上、好ましくは１０５～１０１２ｃｅｌｌｓ／ｍｇ、より好ましくは１０６～１０１１ｃｅｌｌｓ／ｍｇである。 The content of lactic acid bacteria belonging to Lactobacillus pentosus in foods and drinks or pharmaceuticals containing the anti-type I allergy agent of the present invention is appropriately set according to the daily dosage, dosage form, frequency of administration, purpose of use, etc. Ru. For example, the total number of lactic acid bacteria belonging to Lactobacillus pentosus is 10 4 cells/mg or more, preferably 10 5 to 10 per food/drink or pharmaceutical product, although there is no limitation as long as the effects of the present invention are achieved. 12 cells/mg, more preferably 10 6 to 10 11 cells/mg.
また、当該飲食品又は医薬品におけるラクトバチルス・ペントーサスに属する乳酸菌の含有量は、本発明の効果が奏されることを限度として制限されないが、例えば、飲食品又は医薬品あたり、ラクトバチルス・ペントーサスに属する乳酸菌が総菌数して１０４ｃｅｌｌｓ以上、好ましくは１０６～１０１２ｃｅｌｌｓ、より好ましくは１０７～１０１２ｃｅｌｌｓ、更に好ましくは１０８～１０１２ｃｅｌｌｓ、効率良く所望の効果が得られる点から特に好ましくは総菌数として１０８～１０１１ｃｅｌｌｓ、一層好ましくは総菌数として１０９～１０１０ｃｅｌｌｓである。 In addition, the content of lactic acid bacteria belonging to Lactobacillus pentosus in the food/beverage or drug concerned is not limited as long as the effects of the present invention are achieved; The total number of lactic acid bacteria is 10 4 cells or more, preferably 10 6 to 10 12 cells, more preferably 10 7 to 10 12 cells, even more preferably 10 8 to 10 12 cells, and the desired effect can be efficiently obtained. The total number of bacteria is particularly preferably 10 8 to 10 11 cells, and more preferably the total number of bacteria is 10 9 to 10 10 cells.
また、当該飲食品又は医薬品の投与量は、年齢、性別、症状等によって適宜調整されるが、成人１日当たりの投与量として、ラクトバチルス・ペントーサスに属する乳酸菌が総菌数として１０４ｃｅｌｌｓ以上、好ましくは１０６～１０１２ｃｅｌｌｓ、より好ましくは１０７～１０１２ｃｅｌｌｓ、更に好ましくは１０８～１０１２ｃｅｌｌｓ、効率良く所望の効果が得られる点から特に好ましくは１０８～１０１１ｃｅｌｌｓ、一層好ましくは１０９～１０１０ｃｅｌｌｓが例示される。これは、１日に１回又は数回に分けて投与することができる。 In addition, the dosage of the food, drink, or medicine is adjusted appropriately depending on age, gender, symptoms, etc., but the daily dosage for an adult is a total number of lactic acid bacteria belonging to Lactobacillus pentosus of 10 4 cells or more, Preferably 10 6 to 10 12 cells, more preferably 10 7 to 10 12 cells, still more preferably 10 8 to 10 12 cells, particularly preferably 10 8 to 10 11 cells, and even more preferably from the viewpoint of efficiently obtaining the desired effect. A preferred example is 10 9 to 10 10 cells. This can be administered once a day or in several divided doses.
後述する実施例で示すように、ラクトバチルス・ペントーサスに属する乳酸菌は優れたＩ型アレルギーに対する抗アレルギー作用を有するので、抗Ｉ型アレルギー剤、花粉症軽減剤、及びアレルゲンによる不快感の軽減剤の有効成分として使用することができる。また、より詳細には、ラクトバチルス・ペントーサスに属する乳酸菌は優れたＩｇＥ低減作用を有するので、ＩｇＥ低減剤の有効成分として使用することができる。そのため、本発明によれば、ラクトバチルス・ペントーサスに属する乳酸菌を有効成分とする、抗Ｉ型アレルギー剤、花粉症軽減剤、アレルゲンによる不快感の軽減剤、及びＩｇＥ低減剤を提供することができる。また、本発明の抗Ｉ型アレルギー剤等は、食品として使用し得るラクトバチルス・ペントーサスに属する乳酸菌を有効成分とするものであり、副作用が無いか又は極めて少ない。さらに、本発明の抗Ｉ型アレルギー剤等は、軽度のアレルギー反応を示す人のＱＯＬ低下を軽減する目的でも使用することができる。 As shown in the examples below, lactic acid bacteria belonging to Lactobacillus pentosus have an excellent antiallergic effect against type I allergy, so they can be used as anti-type I allergy agents, hay fever alleviation agents, and agents for allergen-induced discomfort. Can be used as an active ingredient. More specifically, lactic acid bacteria belonging to Lactobacillus pentosus have an excellent IgE-reducing effect, and therefore can be used as an active ingredient of an IgE-reducing agent. Therefore, according to the present invention, it is possible to provide an anti-type I allergy agent, a hay fever alleviation agent, an allergen-induced discomfort alleviation agent, and an IgE reducing agent that contain lactic acid bacteria belonging to Lactobacillus pentosus as an active ingredient. . Furthermore, the anti-type I allergy agent of the present invention contains lactic acid bacteria belonging to Lactobacillus pentosus as an active ingredient, which can be used as a food, and has no or very few side effects. Furthermore, the anti-type I allergy agent of the present invention can also be used for the purpose of alleviating the decline in QOL of people who exhibit mild allergic reactions.
前述するように、ラクトバチルス・ペントーサスに属する乳酸菌は、優れたＩ型アレルギーに対する抗アレルギー作用、また、より詳細には、優れたＩｇＥ低減作用を有している。このことから、本発明は更に、ラクトバチルス・ペントーサスに属する乳酸菌を用いてＩ型アレルギー症状を緩和する方法、花粉症を軽減する方法、アレルゲンによる不快感を軽減する方法、及びＩｇＥを低減する方法（以下、Ｉ型アレルギー症状を緩和等する方法と表記することもある）を提供する。本発明のＩ型アレルギー症状を緩和等する方法は、Ｉ型アレルギー症状の緩和等を求める動物にラクトバチルス・ペントーサスに属する乳酸菌を投与することにより実施できる。すなわち、本発明のＩ型アレルギー症状を緩和等する方法は、Ｉ型アレルギー症状の緩和等が要求される動物に、前記抗Ｉ型アレルギー剤等、又は前記抗Ｉ型アレルギー剤等を含有する飲食品又は医薬品を摂取させる工程を含む。本発明の方法におけるラクトバチルス・ペントーサスに属する乳酸菌、前記抗Ｉ型アレルギー剤等、前記抗Ｉ型アレルギー剤等を含有する飲食品又は医薬品の投与量、投与回数、投与方法、投与部位等は前述に従う。 As mentioned above, lactic acid bacteria belonging to Lactobacillus pentosus have an excellent antiallergic effect against type I allergies, and more specifically, an excellent IgE reducing effect. Based on this, the present invention further provides a method for alleviating type I allergy symptoms, a method for alleviating hay fever, a method for alleviating discomfort caused by allergens, and a method for reducing IgE using lactic acid bacteria belonging to Lactobacillus pentosus. (hereinafter also referred to as a method for alleviating type I allergy symptoms). The method of alleviating type I allergy symptoms of the present invention can be carried out by administering lactic acid bacteria belonging to Lactobacillus pentosus to an animal seeking alleviation of type I allergy symptoms. That is, the method for alleviating type I allergy symptoms of the present invention involves administering the anti-type I allergy agent, etc., or food and drink containing the anti-type I allergy agent, etc. to an animal in which alleviation of type I allergy symptoms is required. This includes the process of ingesting a product or medicine. In the method of the present invention, the dosage, frequency of administration, method of administration, administration site, etc. of the lactic acid bacteria belonging to Lactobacillus pentosus, the anti-type I allergy agent, etc., and the food, drink, or pharmaceutical product containing the anti-type I allergy agent are as described above. Follow.
以下、実施例を挙げて本発明を説明するが、本発明はこれらの実施例に限定されるものではない。 The present invention will be described below with reference to Examples, but the present invention is not limited to these Examples.
試験例１
＜試験方法＞
・試験動物：７週齢雄性マウスＢＡＬＢ／ｃＣｒＳｌｃ（日本エスエルシー株式会社）
Test example 1
<Test method>
・Test animal: 7-week-old male mouse BALB/cCrSlc (Japan SLC Co., Ltd.)
・試験群：
（１）コントロール群（ＣＲＦ－１粉末食）（ｎ＝８）
（２）ＰｒｅＢ２４０群（０．２５％ｂ０２４０含有ＣＲＦ－１粉末食）（ｎ＝８）
（３）ＰｏｓｔＢ２４０群（０．２５％ｂ０２４０含有ＣＲＦ－１粉末食）（ｎ＝８）
（４）非感作群（ＣＲＦ－１粉末食）（ｎ＝３）
０．２５％ｂ０２４０含有ＣＲＦ－１粉末食は、ＣＲＦ－１粉末１ｋｇに、乳酸菌ラクトバチルスＯＮＲＩＣｂ０２４０原末２．５ｇを混合して調製した。なお、０．２５％ｂ０２４０は１０ｍｇに相当する。
・Test group:
(1) Control group (CRF-1 powder diet) (n=8)
(2) PreB240 group (CRF-1 powdered food containing 0.25% b0240) (n=8)
(3) PostB240 group (CRF-1 powdered food containing 0.25% b0240) (n=8)
(4) Non-sensitized group (CRF-1 powdered food) (n=3)
A CRF-1 powder food containing 0.25% b0240 was prepared by mixing 1 kg of CRF-1 powder with 2.5 g of lactic acid bacteria Lactobacillus ONRIC b0240 bulk powder. Note that 0.25% b0240 corresponds to 10 mg.
・評価項目：鼻掻き回数、血清ＩｇＥ（総ＩｇＥ）
鼻掻き回数については、観察用ケージ内に入れたマウスに５０ｍｇ／ｍＬ ＯＶＡ溶液を点鼻投与し、その直後からマウスの行動をビデオカメラで撮影した。鼻掻き回数は、マウスが前肢で鼻部を引っ掻いた行動のみを鼻掻き行動として計数した。なお、撮影画像がどの群であるかが分からないよう盲検化し、鼻掻き回数データ固定後に開錠した。
血清ＩｇＥ（総ＩｇＥ）については、Mouse OptEIA ELISA Set Mouse IgEを用い、キットの説明書に従い実施した。
・Evaluation items: Number of nose scratches, serum IgE (total IgE)
Regarding the number of nose scratches, a 50 mg/mL OVA solution was nasally administered to a mouse placed in an observation cage, and immediately thereafter, the behavior of the mouse was photographed with a video camera. The number of nose scratches was counted only when the mouse scratched its nose with its forelimbs. The subjects were blinded so that it was not known which group the photographed images belonged to, and the lock was opened after fixing the data on the number of nose scratches.
Serum IgE (total IgE) was measured using Mouse OptEIA ELISA Set Mouse IgE according to the kit instructions.
・試験スケジュール：図１に示すスケジュールにてＯＶＡによる感作及びラクトバチルスＯＮＲＩＣｂ０２４０の投与を行った。
初回感作は，マウスに腹腔投与用感作液（ＯＶＡ０．５ｍｇ＋水酸化アルミニウムゲル５ｍｇ＋百日咳毒素１．５μｇ／ｍＬ溶液）０．２ｍＬを腹腔内投与した。感作２回目は皮下投与用感作液（ＯＶＡ０．５ｍｇ／ｍＬ溶液）０．１ｍＬを背部皮下投与した。症状惹起は、惹起日午前中に、局所感作として１日１回の頻度で惹起用抗原投与液（ＯＶＡ５０ｍｇ／ｍＬ溶液）を、無麻酔下で保定したマウス両側鼻腔に２μＬずつ、合計４μＬを点鼻投与し、鼻アレルギー症状を惹起させた。非感作群では生理食塩水を感作群と同様の方法で投与した。ラクトバチルスＯＮＲＩＣｂ０２４０の投与は、初回感作以前から０．２５％ｂ０２４０含有ＣＲＦ－１粉末食を給餌する群（ＰｒｅＢ２４０群）と感作２回目翌日から給餌する群（ＰｏｓｔＢ２４０）を設定した。対照群は、ＣＲＦ－１粉末食を給餌した。
- Test schedule: Sensitization with OVA and administration of Lactobacillus ONRICb0240 were performed according to the schedule shown in FIG.
For the initial sensitization, 0.2 mL of a sensitizing solution for intraperitoneal administration (OVA 0.5 mg + aluminum hydroxide gel 5 mg + pertussis toxin 1.5 μg/mL solution) was administered intraperitoneally to the mouse. For the second sensitization, 0.1 mL of a sensitizing solution for subcutaneous administration (OVA 0.5 mg/mL solution) was subcutaneously administered to the back. To induce symptoms, in the morning of the day of challenge, a total of 4 μL of the antigen administration solution (OVA 50 mg/mL solution) was administered once a day as local sensitization to both nasal cavities of mice kept under anesthesia. The drug was administered nasally to induce nasal allergy symptoms. In the non-sensitized group, physiological saline was administered in the same manner as in the sensitized group. Lactobacillus ONRICb0240 was administered to a group that was fed a powdered CRF-1 diet containing 0.25% b0240 before the first sensitization (PreB240 group) and a group that was fed from the day after the second sensitization (PostB240). The control group was fed a CRF-1 powder diet.
＜試験結果＞
試験期間中の体重推移の結果を図２に、鼻掻き回数の結果を図３に、血清ＩｇＥ濃度の結果を図４に示す。
<Test results>
Figure 2 shows the results of the change in body weight during the test period, Figure 3 shows the results of the number of nose scratches, and Figure 4 shows the results of the serum IgE concentration.
図２から試験期間中に平均体重が減少した群は無かった。図３からラクトバチルスＯＮＲＩＣｂ０２４０摂取によりアレルギー性鼻炎モデルマウスの鼻掻き回数が減少していた。図４からＰｒｅＢ２４０群について血清ＩｇＥは減少傾向を認めた。 From FIG. 2, there was no group whose average body weight decreased during the test period. As shown in FIG. 3, the number of nose scratches in allergic rhinitis model mice was reduced by ingestion of Lactobacillus ONRICb0240. As shown in FIG. 4, serum IgE tended to decrease in the PreB240 group.
試験例２
＜試験方法＞
・被験者
被験者は、ＮＰＯ日本健康増進支援機構によって２０～６５歳の間の健康なボランティアから募集された。全てのボランティアは、スギ花粉によると思われるアレルギー症状を経験していた。書面によるインフォームドコンセントを与えたボランティアは、次の選択基準に従い試験責任医師によって試験に適切であると判断され、被験者として登録された。１）同意時に２０～６５歳であること；２）試験に参加するために書面による同意を提供したこと；３）スギ花粉飛散時期以外にアレルギー性鼻炎の症状を示さず、投薬が必要でなかったこと；４）スギ花粉特異的ＩｇＥ値が≧０．７ＵＡ／ｍＬであること；及び５）事前診断後に試験責任医師によって試験に適切であると判断されたこと。
Test example 2
<Test method>
- Subjects Subjects were recruited from healthy volunteers between the ages of 20 and 65 by the NPO Japan Health Promotion Support Organization. All volunteers experienced allergic symptoms believed to be caused by cedar pollen. Volunteers who gave written informed consent were deemed suitable for the study by the study investigator according to the following inclusion criteria and were enrolled as subjects. 1) Be between 20 and 65 years of age at the time of consent; 2) Provide written consent to participate in the study; 3) Show no symptoms of allergic rhinitis outside of the cedar pollen season and do not require medication. 4) The cedar pollen-specific IgE value was ≧0.7 UA/mL; and 5) The subject was judged to be suitable for the study by the study director after a preliminary diagnosis.
除外基準は以下のとおりであった。１）妊娠中又は妊娠している可能性のある女性、及び試験中に妊娠を計画している女性；２）授乳中の女性；３）重度の呼吸器疾患、心臓病、肝臓病、腎臓病、糖尿病、自己免疫性疾患又はこれらの疾患の既往歴を有する被験者；４）急性鼻炎、副鼻腔炎又は肥厚性鼻炎を有する被験者；５）気管支喘息を有する被験者；６）試験の結果に影響し得る治療（舌下の免疫療法など）を受けている被験者；７）抗ヒスタミン剤、抗ロイコトリエン薬、鼻内噴霧ステロイド、経口ステロイド又は免疫療法を受けている被験者；８）牛乳アレルギーを有する被験者；９）乳酸菌が豊富な食品（キムチ、ピクルス、飲料、ヨーグルト、特定保健用食品、機能性表示食品、又はサプリメント）又はプロバイオティクス含有医薬品を定期的に摂取した被験者；１０）抗アレルギー食品を定期的に消費した被験者；１１）事前診断の１２週間以内に４００ｍＬの血液又は４週間以内に２００ｍＬの血液を提供した被験者；及び１２）主任試験責任医師及び主任研究者によって試験に不適切であると考えられた被験者。 Exclusion criteria were as follows. 1) Women who are pregnant or may become pregnant, and women who plan to become pregnant during the study; 2) Women who are breastfeeding; 3) Severe respiratory, heart, liver, or kidney disease. , diabetes, autoimmune disease, or a history of these diseases; 4) subjects with acute rhinitis, sinusitis, or hypertrophic rhinitis; 5) subjects with bronchial asthma; 6) subjects that may affect the results of the study. 7) Subjects receiving antihistamines, antileukotrienes, nasal spray steroids, oral steroids or immunotherapy; 8) Subjects with cow's milk allergy; 9) Subjects who regularly consumed foods rich in lactic acid bacteria (kimchi, pickles, drinks, yogurt, foods for specified health uses, foods with functional claims, or supplements) or medicines containing probiotics; 10) Regularly consumed anti-allergic foods subjects who have consumed; 11) subjects who have donated 400 mL of blood within 12 weeks or 200 mL of blood within 4 weeks of pre-diagnosis; and 12) considered unsuitable for the study by the principal investigator and principal investigator. subjects.
・試験デザイン
試験は、無作為化二重盲検プラセボ対照試験としてデザインされ、２０１８年９月と１２月の間にＮＰＯ日本健康増進支援機構（和歌山、日本）によって実施された。スギ花粉曝露を含む事前検診を受けた５０人のボランティアからインフォームドコンセントを得た。スギ花粉曝露については、ボランティアは曝露施設に入り、３時間、スギ花粉８，０００個／ｍ３に曝露され、その間、３０分ごとに症状を記録した。被験者は、選択基準と除外基準の両方に基づき選ばれ、年齢及び男女比により調整された層別無作為化によって２群に分けられた。１）プラセボ錠剤摂取群（プラセボ群）、２）２０億個のＢ２４０含有錠剤摂取群（Ｂ２４０群）。被験者は、８週間連続して毎日朝食前に１粒の錠剤を摂取し、試験期間中毎日、錠剤の摂取と健康状態を自己評価した。４週目及び８週目において、スギ花粉曝露が、事前検診と同じ方法で行なわれた（図５）。
Study design The study was designed as a randomized, double-blind, placebo-controlled trial and was conducted by the NPO Japan Health Promotion Organization (Wakayama, Japan) between September and December 2018. Informed consent was obtained from 50 volunteers who underwent a preliminary examination including exposure to cedar pollen. For cedar pollen exposure, volunteers entered the exposure facility and were exposed to 8,000 cedar pollen/ m3 for 3 hours, during which symptoms were recorded every 30 minutes. Subjects were selected based on both inclusion and exclusion criteria and were divided into two groups by stratified randomization adjusted for age and sex ratio. 1) a group taking placebo tablets (placebo group), 2) a group taking 2 billion B240-containing tablets (B240 group). Subjects ingested one tablet before breakfast every day for eight consecutive weeks, and self-evaluated their tablet intake and health status every day during the study period. At the 4th and 8th week, cedar pollen exposure was performed in the same manner as the pre-screening (Figure 5).
・試験食品
加熱殺菌されたＢ２４０を含有する錠剤は次のように調製した。Ｂ２４０細胞を培養し、繰り返し洗浄し、殺菌、凍結乾燥を行い、２０億個のＢ２４０細胞を含有する乳糖錠剤を作製した。Ｂ２４０を含まない乳糖錠剤はプラセボとして使用した。２つの錠剤は、大きさ、味及び外観において判別不能だった。
- Test food Tablets containing heat-sterilized B240 were prepared as follows. B240 cells were cultured, washed repeatedly, sterilized, and lyophilized to produce lactose tablets containing 2 billion B240 cells. Lactose tablets without B240 were used as a placebo. The two tablets were indistinguishable in size, taste and appearance.
・測定
評価項目は、みずっぱな、くしゃみ、鼻づまり、鼻のかゆみ、目のかゆみ、なみだめ、鼻水を拭くために使用されたティッシュペーパーの枚数及び総括的な顔スケールを含むアレルギー症状であり、環境曝露施設での３時間のスギ花粉への曝露によって引き起こされ、ＪＲＱＬＱ Ｎｏ．１に基づいたアンケートを参考にして評価された。被験者は、曝露施設に入る前に、症状の重症度、鼻を拭くために使用されたティッシュペーパーの枚数、及び総括的状態を自ら記録した。スギ花粉への曝露中に、アレルギー症状とＱＯＬを評価するために一般的に使用されるＪＲＱＬＱ Ｎｏ．１(1)を参考にして作成されたアンケートを使用して、３０分毎に症状を記録した。ＪＲＱＬＱ Ｎｏ．１は、一般にアレルギー症状、ＱＯＬ、ＱＯＬとアレルギー症状を組み合わせた総括的な顔スコアに関する質問からなる(2)。アンケートは、５段階評価で７つのパラメーター（みずっぱな、くしゃみ、鼻づまり、鼻のかゆみ、目のかゆみ、なみだめ、及び総括的な顔スケール）を含んでいる。鼻水を拭くために使用されたティッシュペーパーの枚数は、パラメーターに追加した（図６）。
(1) Okubo K, Kurono Y, Ichimura K, Enomoto T, Okamoto Y, Kawauchi H et al, Japanese guidelines for allergic rhinitis 2017. Allergol Int2017; 66: 205-19.
(2) Okuda M, Ohkubo K, Goto M, Okamoto Y, Konno A, Baba K, et al. Japanese allergic rhinitis QOL questionnaire. Jpn J Allergology 2003; 52(suppl 1): 21-56.
・Measurement The evaluation items were allergy symptoms including sneezing, stuffy nose, itchy nose, itchy eyes, sore throat, number of tissues used to wipe the runny nose, and overall facial scale. , caused by exposure to cedar pollen for 3 hours in an environmental exposure facility, JRQLQ No. The evaluation was based on a questionnaire based on 1. Subjects self-recorded the severity of symptoms, number of tissues used to wipe the nose, and general condition prior to entering the exposure facility. JRQLQ No. 1, which is commonly used to assess allergic symptoms and QOL during exposure to cedar pollen. Symptoms were recorded every 30 minutes using a questionnaire created with reference to 1 (1) . JRQLQ No. 1 generally consists of questions regarding allergic symptoms, QOL, and a comprehensive facial score that combines QOL and allergic symptoms (2) . The questionnaire includes seven parameters (wateriness, sneezing, stuffy nose, itchy nose, itchy eyes, sluggishness, and general face scale) on a five-point scale. The number of tissues used to wipe the mucus was added to the parameters (Figure 6).
(1) Okubo K, Kurono Y, Ichimura K, Enomoto T, Okamoto Y, Kawauchi H et al, Japanese guidelines for allergic rhinitis 2017. Allergol Int2017; 66: 205-19.
(2) Okuda M, Ohkubo K, Goto M, Okamoto Y, Konno A, Baba K, et al. Japanese allergic rhinitis QOL questionnaire. Jpn J Allergology 2003; 52(suppl 1): 21-56.
・統計分析
全てのデータは平均又は最小二乗平均±ＳＥＭ又はＳＤとして表し、有意水準が５％の両側検定を統計分析に使用した。
-Statistical analysis All data are expressed as mean or least squares mean ± SEM or SD, and a two-tailed test with a significance level of 5% was used for statistical analysis.
曝露３時間の増分スコアは、３時間の間の症状の重症度を示すために、０、４及び８週で、曲線下面積（ＡＵＣ）として合計した。群間で有意差を示したスコアについては、曝露３時間の変化を参考のために示した。 The 3-hour exposure incremental scores were summed as the area under the curve (AUC) at 0, 4, and 8 weeks to indicate the severity of symptoms during the 3-hour period. For scores that showed significant differences between groups, changes over 3 hours of exposure are shown for reference.
データは、０週値（基準値）を共変量とし、各症状を従属変数とし、群、週及び週ごとの群を独立変数として、混合モデルを用いた分散分析による経時測定データ解析を実施した。各週での２群間の統計的な相違は単純主効果テストを実施した。総括的な顔スケールと各症状スコアの間の単純相関は、ピアソンの相関係数を使用して分析し、各症状が総括的な顔スケールにどの程度影響したかを調査するために逐次重回帰分析を行った。変数は、総括的な顔スケール以外の７つの症状を選択した。８週の３時間曝露後の各症状のスコアから、０週のスコアを引いたデルタ値は、対応のないｔ検定（ｖｓプラセボ）を使用して群間で比較した。両側１０％未満を傾向差ありと設定した。統計分析はＳＡＳバージョン９．４(SAS Institute Japan Ltd.)統計ソフトウェアを使用して行った。 The data were analyzed over time by analysis of variance using a mixed model, with week 0 values (reference values) as covariates, each symptom as dependent variable, and group, week, and weekly group as independent variables. . Statistical differences between the two groups at each week were determined using a simple main effects test. Simple correlations between the global face scale and each symptom score were analyzed using Pearson's correlation coefficient and sequential multiple regression to investigate the extent to which each symptom influenced the global face scale. Analysis was carried out. Seven symptoms other than the general face scale were selected as variables. The delta value of each symptom score after the 3-hour exposure at week 8 minus the score at week 0 was compared between groups using an unpaired t-test (vs. placebo). A difference of less than 10% on both sides was considered to be a difference in trend. Statistical analysis was performed using SAS version 9.4 (SAS Institute Japan Ltd.) statistical software.
＜試験結果＞
・被験者
図７は試験のフローを示す。書面によるインフォームドコンセントは、５２人の応募者のうちの５０人から得られ、３４人のボランティアが適格基準に基づき選ばれ、試験に登録された。全ての被験者が８週間の試験を完了したので、最大の解析対象集団（ＦＡＳ）は３４人のボランティアを含んだ。曝露前にアレルギー症状及び風邪の症状を示した４人のボランティア（４週で５人のボランティア）を除外した。したがって、３０人のボランティア（４週で２９人のボランティア）が、per-protocol set（ＰＰＳ）として有効性の分析に含まれた。試験食品の摂取率は、＞８５．７％であり、２つの群の性別又は年齢の間に有意差はなかった（表１）。
<Test results>
- Subject Figure 7 shows the flow of the test. Written informed consent was obtained from 50 of the 52 applicants and 34 volunteers were selected based on eligibility criteria and enrolled in the study. The largest analysis population (FAS) included 34 volunteers, as all subjects completed the 8-week study. Four volunteers who showed allergy symptoms and cold symptoms before exposure (5 volunteers at 4 weeks) were excluded. Therefore, 30 volunteers (29 volunteers at 4 weeks) were included in the efficacy analysis as a per-protocol set (PPS). The intake rate of the test food was >85.7%, with no significant difference between gender or age of the two groups (Table 1).
・スギ花粉曝露時の症状スコアのＡＵＣ
ＦＡＳ及びＰＰＳを使用してＢ２４０群とプラセボ群の間の各症状の３時間の合計スコアを比較した。顔スケールスコアは、ＰＰＳとＦＡＳのいずれにおいても、８週において、Ｂ２４０摂取群はプラセボ群より有意に低かった（表２）。０週及び８週における、曝露３時間にわたる顔スケールスコアの変化を図８に示す。
・AUC of symptom score when exposed to cedar pollen
The 3-hour total score for each symptom was compared between the B240 and Placebo groups using FAS and PPS. Face scale scores were significantly lower in the B240 group than in the placebo group at week 8 for both PPS and FAS (Table 2). Changes in face scale scores over 3 hours of exposure at weeks 0 and 8 are shown in Figure 8.
各週での２群間の統計的な相違は単純主効果テストを実施した。
算術平均±SEM (0 w), 最小二乗平均±SEM (4 w, 8 w) *P < 0.05 (vs.プラセボ)
Arithmetic mean ± SEM (0 w), least squares mean ± SEM (4 w, 8 w) *P < 0.05 (vs. placebo)
・顔スケールと各症状に関する曝露３時間の総スコアのＡＵＣの相関
単純相関分析は、３時間の合計の顔スケールスコアが全ての症状と有意に、正の相関関係があることを示した（表３）。さらに、相関係数は鼻づまりで特に高く、水っぱな、鼻のかゆみ、鼻紙枚数に関するものも高かった。
・Correlation between AUC of total score for 3 hours of exposure for face scale and each symptom Simple correlation analysis showed that total face scale score for 3 hours had a significant positive correlation with all symptoms (Table 3). Furthermore, the correlation coefficient was particularly high for stuffy nose, as well as for watery nose, itchy nose, and number of nasal swabs.
・安全性
副作用：分析対象は、３４人の適格とされたボランティアであった。皮膚のかゆみ及び眼瞼膨張（ｎ＝１）、細菌性結膜炎（ｎ＝１）並びに腎盂炎（ｎ＝１）が副作用として定義されたが、重篤では無いと判断された。３人全てのボランティアがＢ２４０摂取群であった。しかしながら、全ての副作用が試験食品摂取前に生じたか又は一時的であったので、試験責任医師は有害事象と試験食品摂取の間に関係はないと判断した。
- Safety Side effects: The subjects for analysis were 34 eligible volunteers. Itching of the skin and eyelid swelling (n=1), bacterial conjunctivitis (n=1), and pyelonephritis (n=1) were defined as side effects, but were not considered serious. All three volunteers were in the B240 intake group. However, because all adverse events occurred before ingestion of the test food or were temporary, the investigator determined that there was no relationship between the adverse events and the ingestion of the test food.
・３時間曝露後の各症状のスコアの比較
８週の３時間曝露後の各症状のスコアから０週のスコアを引いたデルタ（Δ）値は、プラセボ群とＢ２４０群で比較された（表４）。顔スケール、鼻づまり及び鼻水を拭くために使用されたティッシュペーパーの枚数は、プラセボ群と比べて改善する傾向にあった。
・Comparison of scores for each symptom after 3-hour exposure The delta (Δ) value obtained by subtracting the score for week 0 from the score for each symptom after 3-hour exposure at week 8 was compared between the placebo group and the B240 group (Table 4). Facial scale, nasal congestion, and number of tissues used to wipe nasal discharge tended to improve compared to the placebo group.
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| JP2003300887A (en) | 2002-04-05 | 2003-10-21 | Tanglewood Co Ltd | Mixture having antiallergic effect and food product containing the same |
| WO2005019438A1 (en) | 2003-08-21 | 2005-03-03 | Otsuka Pharmaceutical Co., Ltd. | Lactic acid bacteria having mucosal immunopotentiation effect |
| JP2006321786A (en) | 2005-04-21 | 2006-11-30 | Hokuren Federation Of Agricult Coop:The | Allergy-inhibiting composition, allergy-inhibiting food, and allergy inhibitor |
| WO2008023665A1 (en) | 2006-08-21 | 2008-02-28 | Sapporo Breweries Limited | Bacterial strain having anti-allergic activity and immunostimulating activity, and beverage, food, anti-allergic agent and immunostimulating agent comprising the bacterial strain |
| JP2011004620A (en) | 2009-06-23 | 2011-01-13 | Tokai Igaku Kensa Kenkyusho:Kk | Genus lactobacillus strain and food and medicine containing the strain |
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| JP2003300887A (en) | 2002-04-05 | 2003-10-21 | Tanglewood Co Ltd | Mixture having antiallergic effect and food product containing the same |
| WO2005019438A1 (en) | 2003-08-21 | 2005-03-03 | Otsuka Pharmaceutical Co., Ltd. | Lactic acid bacteria having mucosal immunopotentiation effect |
| JP2006321786A (en) | 2005-04-21 | 2006-11-30 | Hokuren Federation Of Agricult Coop:The | Allergy-inhibiting composition, allergy-inhibiting food, and allergy inhibitor |
| WO2008023665A1 (en) | 2006-08-21 | 2008-02-28 | Sapporo Breweries Limited | Bacterial strain having anti-allergic activity and immunostimulating activity, and beverage, food, anti-allergic agent and immunostimulating agent comprising the bacterial strain |
| JP2011004620A (en) | 2009-06-23 | 2011-01-13 | Tokai Igaku Kensa Kenkyusho:Kk | Genus lactobacillus strain and food and medicine containing the strain |
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