JP3921175B2 - Composition for prevention and / or treatment of oral disease - Google Patents
Composition for prevention and / or treatment of oral disease Download PDFInfo
- Publication number
- JP3921175B2 JP3921175B2 JP2002578985A JP2002578985A JP3921175B2 JP 3921175 B2 JP3921175 B2 JP 3921175B2 JP 2002578985 A JP2002578985 A JP 2002578985A JP 2002578985 A JP2002578985 A JP 2002578985A JP 3921175 B2 JP3921175 B2 JP 3921175B2
- Authority
- JP
- Japan
- Prior art keywords
- extract
- lactic acid
- medium
- xylitol
- acid bacterium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
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- 208000030194 mouth disease Diseases 0.000 title description 14
- 230000002265 prevention Effects 0.000 title description 5
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- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 claims description 67
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Description
技術分野
本発明は、口腔内疾患の予防及び／又は治療用組成物に関する。
背景技術
乳酸菌が人の健康に有用であることは広く知られた事実であり、特に消化管内において、有害菌の増殖を抑制するなどの作用を有することから整腸薬として医薬品にも配合されている。近年、整腸作用とは別に乳酸菌の有用性が明らかにされつつある。その中で、虫歯、歯周病等の口腔内疾患に対する有用性を示すデータも公開されている。特開昭６１−９１１２６号公報には各種乳酸菌の滅菌菌体あるいは水抽出物の歯周病等の代表的原因菌であるバクテロイデス（Ｂａｃｔｅｒｏｉｄｅｓ）属細菌に対する有効性が示されているが、これは滅菌菌体を用いたものであり、生きた乳酸菌の口腔内疾患への応用は示されていない。
また、ＷＯ９９／０７８２６によれば、ある種の特定の乳酸菌が虫歯、歯周病等の口腔内疾患に有効であるとのデータが開示されているが、虫歯、歯周病等の口腔内疾患に対する効率的な利用形態や実施方法に関する技術は未だ解決されていない。例えば、虫歯、歯周病等の口腔内疾患に対する利用形態として、菌液あるいは乳酸菌粉末の塗布をするなどの方法が考えられるが、人が容易にこの方法を実施するには、時間帯、場所において制限がある。また、このような手段では、期待される効果は一時的であり、効率的な利用形態としては充分とはいえない。また、キシリトールが虫歯の発生に抑制的に働くことは広く知られた事実であり、キシリトール入りチューインガムなどの口腔用組成物が多く市販されているが、その虫歯抑制効果は充分とはいえない。
発明の開示
従って、本発明の目的は、口腔内疾患の予防、治療に、殺菌剤などの塗布などに比較して安全であり、何時でも手軽に利用でき、かつ、有効成分が口腔内で持続的に溶出される口腔内疾患の予防及び／又は治療用組成物を提供することである。
本発明の他の目的は、従来のキシリトール含有組成物と比較して、口腔内疾患の予防、治療に、より有効な口腔内疾患の予防及び／又は治療用組成物を提供することである。
本発明者は、上記課題を解決するために鋭意研究を行った結果、生きた乳酸菌、該乳酸菌含有物、該乳酸菌培養ろ液、又はその処理物が、虫歯の原因とされるストレプトコッカス・ミュータンス（Ｓｔｒｅｐｔｏｃｏｃｃｕｓ ｍｕｔａｎｓ）等の虫歯菌や、歯周病の原因とされるポルフィロモナス・ジンジバリス（Ｐｏｒｐｈｙｒｏｍｏｎａｓ ｇｉｎｇｉｖａｌｉｓ）、アクチノバチルス・アクチノミセテムコミタンス（Ａｃｔｉｎｏｂａｃｉｌｌｕｓ ａｃｔｉｎｏｍｙｃｅｔｅｍｃｏｍｉｔａｎｓ）、フソバクテリウム・ヌクレアタム（Ｆｕｓｏｂａｃｔｅｒｉｕｍ ｎｕｃｌｅａｔｕｍ）、プレボテラ・インターメディア（Ｐｒｅｖｏｔｅｌｌａ ｉｎｔｅｒｍｅｄｉａ）等の歯周病菌の増殖に対し、抑制効果を示すこと、さらにまた、驚くべきことに、ストレプトコッカス・ミュータンスやポルフィロモナス・ジンジバリス、アクチノバチルス・アクチノミセテムコミタンス、フソバクテリウム・ヌクレアタム、プレボテラ・インターメディアの増殖に対し、キシリトール等の甘味料や砂糖等の糖類、各種植物抽出物は単独ではほとんど影響しないが、生きた乳酸菌、該乳酸菌含有物、該乳酸菌培養ろ液、又はその処理物にこれらの甘味料や糖類、各種植物抽出物を配合することで、生きた乳酸菌、該乳酸菌含有物、該乳酸菌培養ろ液、又はその処理物単独よりも際だった、これら口腔病原菌の増殖抑制効果を示すことを発見した。本発明はこれらの発見に基づいて完成されたものである。
すなわち本発明は、生きた乳酸菌、該乳酸菌含有物、該乳酸菌培養ろ液、及びその処理物からなる群から選ばれた少なくとも１種（以下、これらの成分を「乳酸菌成分」又は単に「乳酸菌」と呼ぶこともある）と、キシリトール、ソルビトール、エリスリトール、トレハロース、マルチトール、マンニトール、パラチノース、還元乳糖、ショ糖、果糖、ブドウ糖、フラクトオリゴ糖、アスパルテーム、エゾウコギ抽出物、ドクダミ抽出物、紅麹、ウコン色素、カリン抽出物、霊芝抽出物、サンザシ抽出物、シイタケ抽出物、月桃葉抽出物、ステビア抽出物、ハス胚芽抽出物、ラカンカ抽出物、緑茶抽出物、マリアアザミ抽出物、紫玄米抽出物、酵素分解甘草、黄杞葉抽出物、イチョウ抽出物、ルイボス茶抽出物、ヨモギ抽出物、グリチルリチン、ギムネマ抽出物、及び刺梨抽出物からなる群から選ばれた少なくとも１種（以下、これらの成分を「添加物質」又は「添加物」と呼ぶこともある）とを含有する口腔内疾患の予防及び／又は治療用組成物を提供するものである。
本発明に使用される抽出物は、水、含水アルコール（特にエタノール）、アルコール（特にエタノール）により、室温〜溶媒の沸騰温度までの温度で抽出された抽出物である。
発明を実施するための最良の形態
本発明に用いる乳酸菌としては特に制限はないが、ラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）属の乳酸菌、特にヒト口腔内常在菌であるラクトバチルス・サリバリウス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｌｉｖａｒｉｕｓ）、腸管内常在菌であるラクトバチルス・アシドフィルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ａｃｉｄｏｐｈｉｌｌｕｓ）、ラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）、ラクトバチルス・ラムノーサス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｒｈａｍｎｏｓｕｓ）及びラクトバチルス・ジョンソニー（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｊｏｈｎｓｏｎｉｉ）などが安全性の面でも好適であり、より好ましくは口腔内常在菌であるラクトバチルス・サリバリウスである。これら乳酸菌は、本発明の口腔用組成物の一日摂取量あたり、乳酸菌数で、好ましくは１００万個から１０００億個、より好ましくは１億個から１０００億個、最も好ましくは１０億個から１０００億個を配合することが望ましい。
生きた乳酸菌としては、生菌体、湿潤菌、乾燥菌等が適宜使用可能である。乳酸菌含有物としては、乳酸菌懸濁液；乳酸菌培養物（菌体、培養上清液、培地成分を含む）；乳酸菌飲料、酸乳、ヨーグルト等乳酸菌発酵した飲食品からなる乳酸菌発酵乳；等が挙げられる。乳酸菌培養ろ液としては、乳酸菌培養物から乳酸菌を除去した培養ろ液が挙げられる。処理物としては、乳酸菌、乳酸菌含有物、乳酸菌培養ろ液の濃縮物、ペースト化物、乾燥物（噴霧乾燥物、凍結乾燥物、真空乾燥物、ドラム乾燥物等）、液状物、希釈物等が挙げられる。
本発明の組成物中の乳酸菌成分の配合量は、任意でよいが、使用目的（予防又は治療）に応じて適宜定めればよく、通常、０．０００１〜９０質量％、好ましくは０．００１〜２０質量％、さらに好ましくは０．０１〜１０質量％の範囲が適当である。
しかしながら、長期間に亘って保健上ないし健康維持の目的で摂取する場合には、上記範囲よりも少量であってもよいし、また本有効成分は、安全性について問題がないので、上記範囲よりも多量に使用しても一向にさしつかえない。
本発明の口腔内疾患の予防及び／又は治療用組成物の形態は、乳酸菌成分の少なくとも１種と、添加物質の少なくとも１種を含有するものであれば特に限定されないが、例えば、チューインガム、キャンデー、チョコレート、チュアブル錠、歯磨き等の食品又は口腔用組成物の形態が好ましい例として挙げられる。
本発明の組成物中の添加物質の量は、乾燥乳酸菌を使う場合、目的に応じて、少し倍散した濃厚菌末を少量添加して好ましい菌数にしたり、多めに倍散したうすい菌末を多量添加して好ましい菌数にしたりできるので、幅が広い。例えば、キシリトールの場合、好ましくは０．１〜９５．０質量％、さらに好ましくは１〜９０．０質量％、最も好ましくは１０〜６５質量％である。
また、本発明の組成物中の乳酸菌成分と添加物質の質量比率は、例えば、乳酸菌とキシリトールの場合、好ましくは１：５〜１０００である。
食品や口腔用組成物、例えばチューインガムやチョコレートへの生きた菌体の配合には、凍結乾燥した菌体又はその倍散物が利用できる。凍結乾燥菌体の倍散物を調製するには、例えばデンプン、乳糖、白糖などが利用できる。好ましくはデンプンや乳糖で、１ｇ当たりの生菌数を１００億個から２０００億個に希釈したもの（乳酸菌倍散物）が配合操作の面で適している。
チューインガムへの配合には、ガムベースに、例えばキシリトール、ソルビトール、マンニトール、エリスリトール、トレハロース、マルチトール、砂糖、ブドウ糖、アスパルテーム等の甘味料やオリゴ糖、デキストリン等で上記乳酸菌倍散物を希釈したものを加えれば良く、甘味料等の添加物をガムベースに加えた後、乳酸菌倍散物を加えても良い。甘味料としては、特に虫歯に対する有用性が知られているキシリトールとの配合が効果の面で相乗効果が期待でき、より好ましい。また、必要に応じて香料等を加えても何ら問題はない。これらの混合物を練合する温度は、好ましくは４０から８０℃、より好ましくは４０から６０℃、最も好ましくは５０から６０℃が適している。
ガムベース、乳酸菌倍散物、甘味料等をこのように練合したものは、通常の方法により、板ガム、粒ガム等に成型して供することができる。このように調製した乳酸菌入りチューインガム組成物は、配合時の生菌数に対して５０％程度の高い生菌回収率を示し、かつ、溶出試験において良好な徐放性を示す。
チョコレートへの配合には、カカオマス及びココアバターの混合物を４０から６０℃で液化させた後、例えばキシリトール、ソルビトール、マンニトール、エリスリトール、トレハロース、マルチトール、砂糖、ブドウ糖、アスパルテーム等の甘味料やオリゴ糖、デキストリン等で上記乳酸菌倍散物を希釈したものを加えれば良く、甘味料等の添加物を、液化させたカカオマス及びココアバターの混合物に加えた後、乳酸菌倍散物を加えても良い。甘味料としては、特に虫歯に対する有用性が知られているキシリトールとの配合が効果の面で相乗効果が期待でき、より好ましい。また、必要に応じて香料等を加えても何ら問題はない。このように調製した乳酸菌入りチョコレート組成物は、配合時の生菌数に対して１００％近い生菌回収率を示す。
本発明の口腔内疾患の予防及び／又は治療用組成物を適用するのに好適な口腔内疾患の例としては、虫歯、歯周病が挙げられる。
以下に実施例、試験例を示し、より詳細に本発明を説明するが、本発明はこれらに限定されないことは言うまでもない。なおこの明細書中「％」は、他に明記しない限り「質量％」である。
使用した乳酸菌株は以下の通りである。
Ｌｓ：ラクトバチルス・サリバリウスＷＢ２１株（ＦＥＲＭ Ｐ−１７９９１号（＝ＦＥＲＭ ＢＰ−７７９２）、ラクトバシラス・サリバリウスＷＢ２１株は、ＦＥＲＭ ＢＰ−７７９２（寄託日：平成１２年８月１４日）として〒３０５−８５６６茨城県つくば市東１丁目１番地１中央第６の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に寄託されている。
Ｌａ：ラクトバチルス・アシドフィルスＷＢ２００１株（わかもと整腸薬配合菌株）
ＧＧ：ラクトバチルス・ラムノーサスＧＧ株（ＡＴＣＣ５３１０３）
Ｌａ１：ラクトバチルス・ジョンソニーＬａ１株（ＣＮＣＭ Ｉ−１２２５）
実施例１ チューインガム組成物１の製造
恒温槽で３０分間６０℃に加温したガムベース７ｇにキシリトール（東和化成製）９ｇおよびマンニトール（東和化成製）４ｇを加え２分間練合し、再び６０℃にて１０分間加温した。これにラクトバチルス・アシドフィルス菌倍散物（１ｇあたり１．３×１０１１個の生菌を含む）０．０８ｇを加え２分間練合した後、再び６０℃にて１０分間加温した。その後、同操作（２分間練合、１０分間加温）を４回繰り返し充分に練合した。得られた練合物を放冷する過程で、分割、圧延して厚さ約１ｍｍの板状に成型し組成物１を得た。
実施例２ チューインガム組成物２の製造
ラクトバチルス・アシドフィルス菌倍散物の代わりにラクトバチルス・サリバリウス菌倍散物（１ｇあたり３．１６×１０１０個の生菌を含む）０．３５ｇを用い、実施例１と同様に操作し組成物２を得た。
実施例３ チューインガム組成物３の製造
マンニトールの代わりにソルビトール４ｇを用い、実施例２と同様に操作し組成物３を得た。
実施例４ チューインガム組成物１の生菌数測定
本発明の組成物の生菌数測定法は以下の通りである。
１．５％の細菌培養用寒天（和光純薬）を含むラクトバシリＭＲＳ培地（ディフコ）を滅菌後４５から４８℃に冷却し、約２０ｍｌずつ３枚のシャーレに分注し寒天平板を作製した。作製した寒天平板に、適当に希釈した試験溶液を０．１ｍｌずつ加え、コンラージ棒で一様に塗布した後、３７℃で４８から７２時間嫌気培養した。各平板に出現したコロニー数を計測し、平均コロニー数を算出後、試料１ｇ中の生菌数を以下の式により求めた。
試料１ｇ中の生菌数＝平均コロニー数×１０×希釈倍率／試料採取量（ｇ）
実施例１で製造したチューインガム組成物１を幅約１ｍｍ、長さ１５ｍｍ以内に細切し、測定用試料とした。これを約１ｇ正確に秤量し、２０ｍｌの試験管に入れ、希釈液１）を正確に１０ｍｌ加え、以下に示す２種の抽出法により乳酸菌の抽出を行なった。
抽出法１；試験管ミキサーで１分間混合後５分間静置した。この操作を８回繰り返し抽出液を得た。
抽出法２；試験管ミキサーで１分間混合後５分間静置した。この操作を５回繰り返した後、更にスパーテルで１５分間練合し抽出液を得た。
抽出法１及び抽出法２で得られた抽出液の上清を各々正確に１ｍｌ量り希釈液を加え正確に１０倍希釈した。この１０倍希釈操作を更に４回繰り返し試験溶液とし、試験例に記載した方法により生菌数を求めた。また、求められた生菌数と添加時の乳酸菌理論量から生菌回収率（歩留まり）を求めた。結果を表１に示した。
希釈液１）の組成；１％ペプトン（ベクトンディッキンソン）、０．２％ツイーン８０（関東化学）０．８％塩化ナトリウム、０．０２％塩化カリウムを含む０．０１Ｍリン酸緩衝液（ｐＨ７．５）
実施例２で製造したチューインガム組成物２を幅約１ｍｍ、長さ１５ｍｍ以内に細切し、測定用試料とした。
これを約１ｇ正確に秤量し、２０ｍｌの試験管に入れ、希釈液１）を正確に１０ｍｌ加え、以下に示す３種の抽出法により乳酸菌の抽出を行なった。
抽出法１；試験管ミキサーで１分間混合後５分間静置した。この操作を８回繰り返し抽出液を得た。
抽出法２；試験管ミキサーで１分間混合後５分間静置した。この操作を５回繰り返した後、更にスパーテルで１５分間練合し抽出液を得た。
抽出法３；試験管ミキサーで１分間混合後５分間静置した。この操作を５回繰り返した後、更に乳鉢と乳棒で１５分間練合し抽出液を得た。
抽出法１、抽出法２及び抽出法３で得られた抽出液について実施例４と同様に操作し生菌数と生菌回収率（歩留まり）を求めた。結果を表２に示した。
実施例３で製造したチューインガム組成物３を幅約１ｍｍ、長さ１５ｍｍ以内に細切し、測定用試料とした。
これを約１ｇ正確に秤量し、２０ｍｌの試験管に入れ、希釈液１）を正確に１０ｍｌ加え、以下に示す抽出法により乳酸菌の抽出を行なった。
抽出法１；試験管ミキサーで１分間混合後５分間静置した。この操作を５回繰り返した後、更にスパーテルで１５分間練合し抽出液を得た。
抽出法１で得られた抽出液について実施例４と同様に操作し生菌数を求めた。結果を表３に示した。
ガムベース７ｇ、キシリトール９ｇ、マンニトール４ｇ及びラクトバチルス・サリバリウス菌倍散物０．３５ｇを用い、実施例１の製造法に準じ、以下に示す条件にてチューインガム組成物を調製した。
チューインガム組成物７−１；練合温度５０℃、練合時間１時間
チューインガム組成物７−２；練合温度５０℃、練合時間３時間
チューインガム組成物７−３；練合温度６０℃、練合時間１時間
チューインガム組成物７−４；練合温度６０℃、練合時間３時間
チューインガム組成物７−５；練合温度８０℃、練合時間１時間
チューインガム組成物７−６；練合温度８０℃、練合時間３時間
これら６種のチューインガム組成物につき各々実施例５記載の抽出法３に従い抽出液を得、生菌数と回収率を求めた。結果を表４に示した。
実施例２に示した製造法により得られたチューインガム組成物２を幅約１ｍｍ、長さ１５ｍｍ以内に細切し、約１ｇ正確に秤量し、２０ｍｌの試験管に入れ、予め３７℃に加温した希釈液１）を１０ｍｌ加えた。この試験管について３７℃水浴中で５分間静置後、試験管ミキサーで１分間攪拌する操作を１５０分間繰り返した。この間、経時的（試験開始３０分、４５分、６０分、９０分、１２０分、１５０分後）に抽出液を０．５ｍｌ採取し、希釈液１）で希釈し試験溶液を得、実施例４に従い生菌数を求めた。
実施例３に示した製造法により得られたチューインガム組成物３についても同様に試験した。結果を表５に示した。
この試験には、ストレプトコッカス・ミュータンス（ＪＣＭ５７０５株、以下Ｓｍ）及びＬｓ、ＧＧ、Ｌａ１の乳酸菌株を用いた。また、用いた培地は以下の通りである。
培地Ａ：グルコースを１％となるように添加したＧＡＭ培地（日水製薬）
培地Ｂ：キシリトールを１％となるように添加した培地Ａ
乳酸菌生菌数測定培地：変法ＬＢＳ寒天培地（光岡知足編、腸内細菌学、４７７頁、朝倉書店、１９９０年）
Ｓｍ生菌数測定培地：バシトラシンを１０μｇ／ｍｌ、グルコースを１％となるように添加したＧＡＭ寒天培地
乳酸菌、Ｓｍを各々培地Ａで３７℃、一晩前培養し各菌液を得た。この菌液５０μｌを５０ｍｌの培地Ａ及び培地Ｂに接種し、３７℃で乳酸菌及びＳｍの単独培養及び混合培養を行い、経時的に培養液を採取した。採取した培養液５０μｌを適当に希釈し、乳酸菌生菌数測定培地及びＳｍ生菌数測定培地に塗布し、３７℃で３日間嫌気培養した。生じたコロニー数を計測し、生菌数を求めた。結果を表６から表８に示した。
試験例２ ポルフィロモナス・ジンジバリスの増殖に対する乳酸菌、乳酸菌及びキシリトールの混合物の効果
この試験では、ポルフィロモナス・ジンジバリス（ＪＣＭ８５２５株、以下Ｐｇ）及び試験例１に示した乳酸菌株を用いた。また、用いた培地は以下の通りである。
培地Ａ：グルコースを１％となるように添加したＧＡＭ培地（日水製薬）
培地Ｂ：キシリトールを１％となるように添加した培地Ａ
Ｐｇ生菌数測定培地：硫酸ゲンタマイシンを１０μｇ／ｍｌとなるように添加したＥＧ寒天培地（光岡知足編、腸内細菌学、４７５頁、朝倉書店、１９９０年）
乳酸菌、Ｐｇを各々培地Ａで３７℃、一晩前培養し各菌液を得た。この菌液５０μｌを５０ｍｌの培地Ａ及び培地Ｂに接種し、３７℃でＰｇの単独培養及び乳酸菌との混合培養を行い、経時的に培養液を採取した。採取した培養液５０μｌを適当に希釈し、Ｐｇ生菌数測定培地に塗布し、３７℃で３日間嫌気培養した。生じたコロニー数を計測し、生菌数を求めた。結果を表９から表１１に示した。
実施例９ チョコレート組成物１の製造
カカオマス４０ｇ、ココアバター２０ｇを５０℃の恒温槽中で１０分間加温しながら、混合、溶解した。キシリトール３０ｇを加えて１分間かき混ぜた後、ラクトバチルス・サリバリウス菌倍散物（１ｇあたり１．３×１０１０個生菌を含む）１０ｇを加えて、さらに１分間混合した。シャーレに分注して室温まで冷やした後、５℃で３時間冷却固化して、チョコレート組成物１を得た。同じ製造法により３ロット製造した。
実施例１０ チョコレート組成物１の生菌数測定
製造したチョコレート組成物１を幅約１ｍｍ長さ１５ｍｍ以内に細切し、測定用試料とした。
約１ｇを正確に秤量し、２０ｍｌの試験管に入れ、希釈液１）を正確に１０ｍｌ加え、試験管ミキサーで攪拌しながら５０℃の恒温槽で５分間加温して、溶解した。この１ｍｌを正確に量り、希釈液を加え正確に１０倍希釈した。この１０倍希釈操作を更に４回繰り返し試験溶液とし、試験例１に記載した方法により生菌数を求めた。また求められた生菌数と添加時の乳酸菌理論量から生菌回収率（歩留まり）を求めた。結果を表１２に示した。
キシリトール（東和化成製）１５０ｇ、レシス（東和化成製）５４ｇ、メチルセルロース（信越化学製）６ｇ、部分α化デンプン（旭化成工業製）４５ｇ、ポリデキストロース（和光純薬製）３０ｇ、ショ糖脂肪酸エステル（三菱化学フーズ製）６ｇ、カスターワックス（日本油脂製）６ｇ、軽質無水ケイ酸（フロイント産業製）３ｇ、ラクトバチルス・サリバリウス菌倍散物（１ｇあたり１．３×１０１１個の生菌を含む）１５ｇをビニール袋中でよく混合した後、打錠機（６Ｂ−２、菊水製作所製）を用いて打錠し、直径１５ｍｍ、重量１０００ｍｇのチュアブル錠を得た。
実施例１２ 徐放性口腔内挿入錠の製造
キシリトール（東和化成製）１５０ｇ、粉末還元麦芽糖（東和化成製）５４ｇ、デキストリン（松谷化学工業製）６０ｇ、グルテン（和光純薬製）１５ｇ、メチルセルロース（信越化学製）６ｇを転動流動層造粒機（ニューマルメライザーＮＱ−Ｌａｂｏ、不二パウダル製）を用いて８０％エタノール溶液を噴霧し造粒した。造粒物を６０℃で１６時間通風乾燥した後に、１８メッシュ篩で整粒し打錠用顆粒を得た。打錠用顆粒２７０ｇにラクトバチルス・サリバリウス菌倍散物（１ｇあたり１．３×１０１１の生菌を含む）１５ｇとカスターワックス（日本油脂製）１５ｇを加えよく混合した後、打錠機（ＲＴ−Ｆ９、菊水製作所製）を用いて打錠し、直径６ｍｍ、重量１００ｍｇの徐放性口腔内挿入錠を得た。
試験例３ ストレプトコッカス・ミュータンス（Ｓｍ）の不溶性グルカン産生に対する乳酸菌、乳酸菌及びキシリトールの混合物の効果
乳酸菌はＬｓを用いた。また、用いた培地は以下のとおりである。
培地Ａ：ショ糖を２％となるように添加したＧＡＭ培地（日水製薬）
培地Ｂ：キシリトールを１％となるように添加した培地Ａ
Ｓｍ、Ｌｓを各々培地Ａで３７℃、一晩前培養し各菌液を得た。この菌液０．１ｍｌを１０ｍｌの培地Ａ及び培地Ｂに接種し、Ｓｍ及びＬｓの単独培養及び混合培養を３７℃で２４時間行った。培養液を遠心分離（８０００ｒｐｍ、２０分）して沈殿（菌体＋不溶性グルカン）を回収し、１０ｍｌのリン酸緩衝生理液（以下、ＰＢＳ）で３回洗浄した。０．１Ｎ ＮａＯＨ ５ｍｌを加えて、不溶性グルカンを溶解し、遠心分離（８０００ｒｐｍ、２０分）して、上清を回収した。この上清中の糖濃度をフェノール硫酸法により測定し、不溶性グルカン産生量を調べた。上清０．２ｍｌに５％フェノール０．２ｍｌ、濃硫酸１ｍｌを加えて、１０分間静置後、混和して、さらに２０分間静置した。反応液の４９２ｎｍの吸光度を測定し、グルコース溶液を用いて作成した検量線に基づき、不溶性グルカン濃度を算出した。結果を表１３に示した。
試験例４ 乳酸菌培養液によるポルフィロモナス・ジンジバリスの増殖抑制効果を増強する物質の探索（１）
キシリトールのように、乳酸菌によるポルフィロモナス・ジンジバリス（Ｐｇ）の増殖抑制効果を増強する物質の探索を行った。用いた乳酸菌はＬｓであり、培地は以下のとおりである。
培地Ａ：グルコースを１％となるように添加したＧＡＭ培地（日水製薬）
培地Ｂ：Ｌｓの培養ろ液を３０％となるように添加した培地Ａ
Ｌｓの培養ろ液は、培地Ａで３７℃、２４時間培養したＬｓの培養液を遠心分離（８０００ｒｐｍ、２０分）して上清を回収し、さらに無菌ろ過して調製したもの。
Ｐｇ生菌数測定培地：硫酸ゲンタマイシンを１０μｇ／ｍｌとなるように添加したＥＧ寒天培地（光岡知足編、腸内細菌学、４７５頁、朝倉書店、１９９０年）
Ｐｇを培地Ａで３７℃、一晩培養した前培養液１００μｌを１０ｍｌの培地Ｂ及び１０％の各種物質を添加した培地Ｂに接種し、３７℃で２４時間培養を行った。培養液を適当に希釈し、Ｐｇ生菌数測定培地に塗布し、３７℃で３日間嫌気培養した。生じたコロニー数を計測し生菌数を求めた。結果を表１４に示した。
試験例５ アクチノバチルス・アクチノミセテムコミタンスの増殖に対する乳酸菌培養液及びキシリトールの効果
アクチノバチルス・アクチノミセテムコミタンス（ＪＣＭ８５７７株、以下Ａａ）を用いた。本試験例に用いた乳酸菌はＬｓであり、培地は以下の通りである。
培地Ａ：ブレインハートインヒュージョン液体培地（ベクトンディッキンソン）
培地Ｂ：Ｌｓの培養ろ液を２５％となるように添加した培地Ａ
Ｌｓの培養ろ液は、グルコースを１％となるように添加したＧＡＭ培地で３７℃、２４時間培養したＬｓの培養液を遠心分離（８０００ｒｐｍ、２０分）して上清を回収し、さらに無菌ろ過して調製したもの。
Ａａ生菌数測定培地：ブレインハートインヒュージョン寒天培地（ベクトンディッキンソン）
Ａａを培地Ａで３７℃、一晩培養した前培養液を生菌数約１０６／ｍｌとなるように１０ｍｌの各培地（培地Ａ、培地Ｂ、３％、７％のキシリトールを添加した培地Ａ及び３％、７％のキシリトールを添加した培地Ｂ）に接種し、３７℃で４８時間微好気培養を行った。培養液を適当に希釈し、Ａａ生菌数測定培地に塗布し、３７℃で３日間嫌気培養した。生じたコロニー数を計測し、生菌数を求めた。結果を表１５に示した。
試験例６ フソバクテリウム・ヌクレアタムの増殖に対する乳酸菌培養液及びキシリトールの効果
フソバクテリウム・ヌクレアタム（ＪＣＭ８５３２Ｔ株、以下Ｆｎ）を用いた。本試験例に用いた乳酸菌はＬｓであり、培地は以下の通りである。
培地Ａ：グルコースを１％となるように添加したＧＡＭ培地（日水製薬）
培地Ｂ：Ｌｓの培養ろ液を５０％となるように添加した培地Ａ
Ｌｓの培養ろ液は、試験例５と同様の方法により調製したもの。
Ｆｎ生菌数測定培地：ブレインハートインヒュージョン寒天培地（ベクトンディッキンソン）
Ｆｎを培地Ａで３７℃、一晩培養した前培養液を生菌数約１０６／ｍｌとなるように１０ｍｌの各培地（培地Ａ、培地Ｂ、３％、１０％のキシリトールを添加した培地Ａ及び３％、１０％のキシリトールを添加した培地Ｂ）に接種し、３７℃で２４時間嫌気培養を行った。培養液を適当に希釈し、Ｆｎ生菌数測定培地に塗布し、３７℃で３日間嫌気培養した。生じたコロニー数を計測し、生菌数を求めた。結果を表１６に示した。
試験例７ ポルフィロモナス・ジンジバリスの口腔細胞への付着に対する乳酸菌培養液およびキシリトールの抑制効果
乳酸菌はＬｓを用いた。Ｌｓの培養ろ液は、グルコースを１％となるように添加したＧＡＭ培地で３７℃、２４時間培養したＬｓの培養液を遠心分離（８０００ｒｐｍ、２０分）して上清を回収し、さらに無菌ろ過後、１Ｎ水酸化ナトリウムで中和したもの。
Ｐｇをグルコースが１％となるように添加したＧＡＭ培地で３７℃、一晩培養した。ＰＢＳで洗浄し、生菌数が１０７／ｍｌになるように以下の各培地（培地Ａ、培地Ｂ、２．５％、５％、１０％のキシリトールを添加した培地Ａ及び２．５％、５％、１０％のキシリトールを添加した培地Ｂ）に懸濁し、細胞付着用のＰｇ懸濁液を調製した。
培地Ａ：ＤＭＥＭ／Ｆ−１２培地（キブコ）
培地Ｂ：Ｌｓの培養ろ液を２５％となるように添加した培地Ａ
口腔細胞は、ヒト口腔培養細胞ＨＯ１−ｕ−１（ＪＣＲＢ０８２８株）を用いた。細胞は、４×１０５ｃｅｌｌｓ／ｗｅｌｌになるように２４穴プレートに播種し、３７℃、５％ＣＯ２条件下で一晩培養し単層を形成させた。ＰＢＳで３回洗浄後、細胞付着用のＰｇ懸濁液を１ｍｌ添加し、３７℃、５％ＣＯ２条件下で１．５時間静置した。細胞をＰＢＳで３回洗浄した後、滅菌水１ｍｌを加え、１０分間室温に静置して、細胞を剥離した。ＰＢＳで適宜希釈した後、細胞に付着したＰｇの生菌数を測定した。測定には５％ウマ脱繊血を含むＥＧ寒天培地を用い、３７℃、嫌気条件下で３日間培養した。結果を表１７に示した。
試験例８ 乳酸菌培養液によるポルフィロモナス・ジンジバリスの増殖抑制効果を増強する物質の探索（２）
キシリトールのように、乳酸菌によるポルフィロモナス・ジンジバリスの増殖抑制効果を増強する物質の探索を行った。本試験例に用いた乳酸菌はＬｓであり、培地は以下の通りである。
培地Ａ：グルコースを１％となるように添加したＧＡＭ培地（日水製薬）
培地Ｂ：Ｌｓの培養ろ液を２５％となるように添加した培地Ａ
Ｌｓの培養ろ液は、培地Ａで３７℃、２４時間培養したＬｓの培養液を遠心分離（８０００ｒｐｍ、２０分）して上清を回収し、さらに無菌ろ過して調製したもの。
Ｐｇ生菌数測定培地：硫酸ゲンタマイシンを１０μｇ／ｍｌとなるように添加したＥＧ寒天培地
Ｐｇを培地Ａで３７℃、一晩培養した前培養液１００μｌを１０ｍｌの培地Ｂ及び０．２〜１０％の各種物質を添加した培地Ｂに接種し、３７℃で９時間培養を行った。培養液を適当に希釈し、Ｐｇ生菌数測定培地に塗布し、３７℃で３日間嫌気培養した。生じたコロニー数を計測し、生菌数を求めた。結果を表１８、表１９に示した。なお表１８、表１９に示す実験は独立に行ったため、表１８の添加物質無添加の場合のＰｇ生菌数と、表１９の添加物質無添加の場合のＰｇ生菌数の数値は異なっている。
各種植物抽出物、紅麹、ウコン色素、酵素分解甘草、グリチルリチンは丸善製薬社製のものを用いた。
試験例９ ポルフィロモナス・ジンジバリスの増殖に対する乳酸菌（ビフィズス菌）培養液及びキシリトールの効果
本試験例に用いた乳酸菌はビフィズス菌の２菌種ビフィドバクテリウム・ロンガム（Ｂｉｆｉｄｏｂａｃｔｅｒｉｕｍ ｌｏｎｇｕｍ ＷＢ１００１株（ＦＥＲＭ Ｐ−１２６１０）、以下Ｂｌ）及びビフィドバクテリウム・ビフィダム（Ｂｉｆｉｄｏｂａｃｔｅｒｉｕｍ ｂｉｆｉｄｕｍ ＷＢ１００３株（ＦＥＲＭ Ｐ−１２６１２）、以下Ｂｂ）であり、培地は以下の通りである。
培地Ａ：グルコースを１％となるように添加したＧＡＭ培地（日水製薬）
培地Ｂ：Ｂｌの培養ろ液を２０％となるように添加した培地Ａ
培地Ｃ：Ｂｂの培養ろ液を３５％となるように添加した培地Ａ
Ｂｌ及びＢｂの培養ろ液は、各菌株を、培地Ａで３７℃２０時間培養した培養液を遠心分離（８０００ｒｐｍ、２０分）して上清を回収し、さらに無菌ろ過して調製したもの。
Ｐｇ生菌数測定培地：硫酸ゲンタマイシンを１０μｇ／ｍｌとなるように添加したＥＧ寒天培地
Ｐｇを培地Ａで３７℃、一晩培養した前培養液１００μｌを１０ｍｌの培地Ｂ、培地Ｃ、１０％のキシリトールを添加した培地Ｂ及び１０％のキシリトールを添加した培地Ｃに接種し、３７℃で９時間培養を行った。培養液を適当に希釈し、Ｐｇ生菌数測定培地に塗布し、３７℃で３日間嫌気培養した。生じたコロニー数を計測し、生菌数を求めた。結果を表２０に示した。
試験例１０ プレボテラ・インターメディアの増殖に対する乳酸菌培養液及びキシリトールの効果
プレボテラ・インターメディア（ＪＣＭ７３６５株、以下Ｐｉ）を用いた。本試験例に用いた乳酸菌はＬｓであり、培地は以下の通りである。
培地Ａ：グルコースを１％となるように添加したＧＡＭ培地（日水製薬）
培地Ｂ：Ｌｓの培養ろ液を５０％となるように添加した培地Ａ
Ｌｓの培養ろ液は、試験例５と同様の方法により調製したもの。
Ｐｉ生菌数測定培地：５％ウマ脱繊血を含むＥＧ寒天培地
Ｐｉを培地Ａで３７℃、一晩培養した前培養液を生菌数約１０７／ｍｌとなるように１０ｍｌの各培地（培地Ａ、培地Ｂ、１０％のキシリトールを添加した培地Ａ及び１０％のキシリトールを添加した培地Ｂ）に接種し、３７℃で２４時間嫌気培養を行った。培養液を適当に希釈し、Ｐｉ生菌数測定培地に塗布し、３７℃で３日間嫌気培養した。生じたコロニー数を計測し、生菌数を求めた。結果を表２１に示した。
産業上の利用可能性
本発明の生きた乳酸菌、該乳酸菌含有物、該乳酸菌培養ろ液、及びその処理物からなる群から選ばれた少なくとも１種と、キシリトール、ソルビトール、エリスリトール、トレハロース、マルチトール、マンニトール、パラチノース、還元乳糖、ショ糖、果糖、ブドウ糖、フラクトオリゴ糖、アスパルテーム、エゾウコギ抽出物、ドクダミ抽出物、紅麹、ウコン色素、カリン抽出物、霊芝抽出物、サンザシ抽出物、シイタケ抽出物、月桃葉抽出物、ステビア抽出物、ハス胚芽抽出物、ラカンカ抽出物、緑茶抽出物、マリアアザミ抽出物、紫玄米抽出物、酵素分解甘草、黄杞葉抽出物、イチョウ抽出物、ルイボス茶抽出物、ヨモギ抽出物、グリチルリチン、ギムネマ抽出物、及び刺梨抽出物からなる群から選ばれた少なくとも１種とを含有する口腔内疾患の予防及び／又は治療用組成物、特にチューインガム組成物は良好な徐放性により、口腔内に持続的に乳酸菌を滞留させることができ、安全で、簡便で、効率的な口腔内疾患の予防と治療に用いることができる。Technical field
The present invention relates to a composition for preventing and / or treating oral diseases.
Background art
It is a well-known fact that lactic acid bacteria are useful for human health, and in particular, in the digestive tract, it has an action such as suppressing the growth of harmful bacteria, and is also incorporated in pharmaceutical products as an intestinal adjuster. In recent years, the usefulness of lactic acid bacteria has been revealed separately from the intestinal regulating action. Among them, data showing usefulness for oral diseases such as dental caries and periodontal diseases are also disclosed. JP-A-61-91126 shows the effectiveness of Bacteroides bacteria, which is a typical causative bacterium such as periodontal disease of various lactic acid bacteria or water extract. It uses sterilized bacterial cells, and application of live lactic acid bacteria to oral diseases is not shown.
Further, according to WO99 / 07826, data indicating that certain specific lactic acid bacteria are effective for oral diseases such as dental caries and periodontal diseases is disclosed, but oral diseases such as dental caries and periodontal diseases are disclosed. The technology regarding the efficient use form and implementation method is still not solved. For example, a method of applying a bacterial solution or lactic acid bacteria powder as a form of use for oral diseases such as tooth decay and periodontal disease is conceivable. There are limitations. Further, with such means, the expected effect is temporary, and it cannot be said that it is sufficient as an efficient usage form. In addition, it is a well-known fact that xylitol acts to suppress the occurrence of dental caries, and many oral compositions such as chewing gum containing xylitol are commercially available, but the effect of inhibiting caries is not sufficient.
Disclosure of the invention
Therefore, the object of the present invention is to prevent and treat oral diseases, which is safer than application of bactericides, etc., and can be easily used at any time, and the active ingredient is dissolved in the oral cavity continuously. It is intended to provide a composition for preventing and / or treating oral diseases.
Another object of the present invention is to provide a composition for the prevention and / or treatment of oral diseases that is more effective for the prevention and treatment of oral diseases as compared with conventional xylitol-containing compositions.
As a result of earnest research to solve the above-mentioned problems, the present inventor has found that a living lactic acid bacterium, the lactic acid bacterium-containing material, the lactic acid bacterium culture filtrate, or a treated product thereof is a Streptococcus mutans that causes dental caries. (Streptococcus mutans) and dental caries bacteria such as Porphyromonas gingivalis, which is the cause of periodontal disease (Porphyromonas gingivalis), Actinobacillus actinomycetemcomitans (Actinobacillus actinomycetemcomitans), Fusobacterium nucleatum (Fusobacterium nucleatum), Prevotella -Inhibiting the growth of periodontal disease bacteria such as intermedia (Prevotella intermedia), In particular, for the growth of Streptococcus mutans, Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Prevotella intermedia, sweeteners such as xylitol, sugars such as sugar, Plant extracts alone have little effect, but live by adding these sweeteners, sugars, and various plant extracts to live lactic acid bacteria, lactic acid bacteria-containing materials, lactic acid bacteria culture filtrates, or processed products thereof. It has been discovered that these oral pathogenic bacteria have a growth-inhibiting effect that is more prominent than lactic acid bacteria, the lactic acid bacteria-containing material, the lactic acid bacteria culture filtrate, or the treated product alone. The present invention has been completed based on these findings.
That is, the present invention is at least one selected from the group consisting of a living lactic acid bacterium, the lactic acid bacterium-containing material, the lactic acid bacterium culture filtrate, and a processed product thereof (hereinafter, these components are referred to as “lactic acid bacterium components” or simply “lactic acid bacteria”). Xylitol, sorbitol, erythritol, trehalose, maltitol, mannitol, palatinose, reduced lactose, sucrose, fructose, glucose, fructooligosaccharides, aspartame, sorghum extract, dokudami extract, red yeast rice, turmeric Pigment, karin extract, reishi extract, hawthorn extract, shiitake extract, moon peach leaf extract, stevia extract, lotus germ extract, rakanka extract, green tea extract, maria thistle extract, purple brown rice extract , Enzymatic degradation licorice, yellow leaves extract, ginkgo biloba extract, rooibos tea extract, mugwort extract, glycyrrhiz Or at least one selected from the group consisting of Gymnema extract and prickly pear extract (hereinafter, these components may be referred to as “additive substances” or “additives”) Prophylactic and / or therapeutic compositions are provided.
The extract used in the present invention is an extract extracted with water, hydrous alcohol (particularly ethanol), or alcohol (particularly ethanol) at a temperature from room temperature to the boiling temperature of the solvent.
BEST MODE FOR CARRYING OUT THE INVENTION
The lactic acid bacterium used in the present invention is not particularly limited, but a lactic acid bacterium belonging to the genus Lactobacillus, in particular, Lactobacillus salivarius which is a human oral resident bacterium, Lactobacillus salivarius which is an resident bacteria in the intestinal tract. Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus rhamnosus, Lactobacillus johnson (Lactobacillus johnis) It is Lactobacillus salivarius which is resident. These lactic acid bacteria are preferably from 1 million to 100 billion, more preferably from 100 to 100 billion, most preferably from 1 billion per day of daily intake of the oral composition of the present invention. It is desirable to blend 100 billion.
As living lactic acid bacteria, viable cells, wet bacteria, dry bacteria and the like can be used as appropriate. Lactic acid bacteria-containing materials include: lactic acid bacteria suspensions; lactic acid bacteria cultures (including cell bodies, culture supernatants and medium components); Can be mentioned. Examples of the lactic acid bacteria culture filtrate include a culture filtrate obtained by removing lactic acid bacteria from a lactic acid bacteria culture. Processed products include lactic acid bacteria, lactic acid bacteria-containing materials, concentrated lactic acid bacteria culture filtrates, pasted products, dried products (spray-dried products, freeze-dried products, vacuum-dried products, drum-dried products, etc.), liquid products, diluted products, etc. Can be mentioned.
The blending amount of the lactic acid bacteria component in the composition of the present invention may be arbitrary, but may be appropriately determined according to the purpose of use (prevention or treatment), and is usually 0.0001 to 90% by mass, preferably 0.001. The range of -20% by mass, more preferably 0.01-10% by mass is appropriate.
However, when ingested for the purpose of health or health maintenance over a long period of time, the amount may be smaller than the above range, and since this active ingredient has no safety problem, Even if it is used in large quantities, there is no problem.
The form of the composition for preventing and / or treating oral diseases of the present invention is not particularly limited as long as it contains at least one lactic acid bacterium component and at least one additive substance. For example, chewing gum and candy Preferred examples include foods such as chocolates, chewable tablets, toothpastes, and oral compositions.
When using dry lactic acid bacteria, the amount of the additive substance in the composition of the present invention may be adjusted to a preferred number by adding a small amount of concentrated powdered powder, or a slightly diluted powdered bacterial powder depending on the purpose. Since a large amount of can be added to obtain a preferable number of bacteria, the range is wide. For example, in the case of xylitol, it is preferably 0.1 to 95.0% by mass, more preferably 1 to 90.0% by mass, and most preferably 10 to 65% by mass.
Moreover, the mass ratio of the lactic acid bacteria component and the additive substance in the composition of the present invention is preferably 1: 5 to 1000 in the case of lactic acid bacteria and xylitol, for example.
Freeze-dried microbial cells or their trifolds can be used for blending live microbial cells in food or oral compositions such as chewing gum and chocolate. For example, starch, lactose, sucrose, and the like can be used to prepare a lyophilized bacterial powder. Preferably, starch or lactose diluted with the number of viable bacteria per gram from 10 billion to 200 billion (lactic acid bacteria triturate) is suitable in terms of blending operation.
For chewing gum formulation, for example, xylitol, sorbitol, mannitol, erythritol, trehalose, maltitol, sugar, glucose, aspartame and other sweeteners such as xylitol, sorbitol, oligosaccharide, dextrin, etc. What is necessary is just to add, and after adding additives, such as a sweetener, to a gum base, you may add a lactic acid bacteria triturate. As a sweetener, a combination with xylitol, which is known to be useful for caries in particular, can be expected to produce a synergistic effect, and is more preferable. Moreover, there is no problem even if a fragrance is added if necessary. The temperature at which these mixtures are kneaded is preferably 40 to 80 ° C, more preferably 40 to 60 ° C, and most preferably 50 to 60 ° C.
What knead | mixed the gum base, the lactic acid bacteria triturate, the sweetener etc. in this way can be shape | molded and used for a board gum, a granule gum, etc. by a normal method. The chewing gum composition containing lactic acid bacteria thus prepared exhibits a high viable cell recovery rate of about 50% with respect to the viable cell count at the time of blending, and exhibits a good sustained release property in an elution test.
For blending into chocolate, a mixture of cacao mass and cocoa butter is liquefied at 40 to 60 ° C. and then sweeteners and oligosaccharides such as xylitol, sorbitol, mannitol, erythritol, trehalose, maltitol, sugar, glucose, aspartame, etc. What is necessary is just to add what diluted the said lactic acid bacteria trituration with dextrin etc., and after adding additives, such as a sweetener, to the mixture of liquefied cacao mass and cocoa butter, you may add lactic acid bacteria trituration. As a sweetener, a combination with xylitol, which is known to be useful for caries in particular, can be expected to produce a synergistic effect, and is more preferable. Moreover, there is no problem even if a fragrance is added if necessary. The chocolate composition containing lactic acid bacteria thus prepared exhibits a viable cell recovery rate of nearly 100% with respect to the viable cell count at the time of blending.
Examples of oral diseases suitable for applying the composition for preventing and / or treating oral diseases of the present invention include dental caries and periodontal diseases.
Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples, but it goes without saying that the present invention is not limited thereto. In this specification, “%” is “% by mass” unless otherwise specified.
The lactic acid strains used are as follows.
Ls: Lactobacillus salivarius WB21 strain (FERM P-17991 (= FERM BP-7792), Lactobacillus salivarius WB21 strain is FERM BP-7792 (Deposit date: August 14, 2000) It is deposited at the patent biological deposit center of the National Institute of Advanced Industrial Science and Technology, 1st, 1st Street, Tsukuba City, Ibaraki Prefecture.
La: Lactobacillus acidophilus WB2001 strain (Wakamoto and intestinal combination drug strain)
GG: Lactobacillus rhamnosus GG strain (ATCC 53103)
La1: Lactobacillus johnsonii La1 strain (CNCM I-1225)
Example 1 Production of Chewing Gum Composition 1
9 g of xylitol (manufactured by Towa Kasei) and 4 g of mannitol (manufactured by Towa Kasei) were added to 7 g of a gum base heated to 60 ° C. for 30 minutes in a thermostatic bath, kneaded for 2 minutes, and heated again at 60 ° C. for 10 minutes. Lactobacillus acidophilus bacteria triturate (1.3 x 10 per gram) 11 After adding 0.08 g (including live bacteria) and kneading for 2 minutes, the mixture was heated again at 60 ° C. for 10 minutes. Thereafter, the same operation (kneading for 2 minutes, heating for 10 minutes) was repeated four times and sufficiently kneaded. In the process of allowing the obtained kneaded product to cool, it was divided and rolled to form a plate having a thickness of about 1 mm to obtain a composition 1.
Example 2 Production of Chewing Gum Composition 2
Instead of Lactobacillus acidophilus triturate, Lactobacillus salivarius triturate (3.16 x 10 per gram) 10 A composition 2 was obtained by operating in the same manner as in Example 1 using 0.35 g).
Example 3 Production of Chewing Gum Composition 3
A composition 3 was obtained in the same manner as in Example 2 except that 4 g of sorbitol was used instead of mannitol.
Example 4 Viable count of chewing gum composition 1
The method for measuring the viable cell count of the composition of the present invention is as follows.
Lactobacillus MRS medium (Difco) containing 1.5% bacterial culture agar (Wako Pure Chemical Industries) was sterilized, cooled to 45 to 48 ° C., and dispensed into three petri dishes of about 20 ml each to prepare agar plates. To the prepared agar plate, 0.1 ml each of an appropriately diluted test solution was added and uniformly applied with a conage bar, followed by anaerobic culture at 37 ° C. for 48 to 72 hours. After counting the number of colonies that appeared on each plate and calculating the average number of colonies, the number of viable bacteria in 1 g of the sample was determined by the following formula.
Number of viable bacteria in 1 g of sample = average number of colonies × 10 × dilution rate / sample collection amount (g)
The chewing gum composition 1 produced in Example 1 was cut into a width of about 1 mm and a length of 15 mm to obtain a measurement sample. About 1 g of this is accurately weighed and placed in a 20 ml test tube. 1) 10 ml was added accurately, and lactic acid bacteria were extracted by the following two extraction methods.
Extraction method 1: mixed for 1 minute with a test tube mixer and then allowed to stand for 5 minutes. This operation was repeated 8 times to obtain an extract.
Extraction method 2: mixed for 1 minute with a test tube mixer and allowed to stand for 5 minutes. This operation was repeated 5 times, and further kneaded with a spatula for 15 minutes to obtain an extract.
The supernatants of the extract obtained by the extraction method 1 and the extraction method 2 were each accurately diluted by 1 ml and diluted exactly 10 times by adding a diluent. This 10-fold dilution operation was further repeated 4 times to obtain a test solution, and the viable cell count was determined by the method described in the test example. Further, the viable cell recovery rate (yield) was determined from the obtained viable cell count and the theoretical amount of lactic acid bacteria at the time of addition. The results are shown in Table 1.
Diluted solution 1) Composition: 1% peptone (Becton Dickinson), 0.2% Tween 80 (Kanto Chemical) 0.01M phosphate buffer (pH 7.5) containing 0.8% sodium chloride and 0.02% potassium chloride
The chewing gum composition 2 produced in Example 2 was cut into a width of about 1 mm and a length of 15 mm to obtain a measurement sample.
About 1 g of this is accurately weighed and placed in a 20 ml test tube. 1) Was exactly 10 ml, and lactic acid bacteria were extracted by the following three extraction methods.
Extraction method 1: mixed for 1 minute with a test tube mixer and then allowed to stand for 5 minutes. This operation was repeated 8 times to obtain an extract.
Extraction method 2: mixed for 1 minute with a test tube mixer and allowed to stand for 5 minutes. This operation was repeated 5 times, and further kneaded with a spatula for 15 minutes to obtain an extract.
Extraction method 3: Mixed for 1 minute with a test tube mixer and allowed to stand for 5 minutes. After repeating this operation five times, the mixture was further kneaded for 15 minutes with a mortar and pestle to obtain an extract.
About the extract obtained by the extraction method 1, the extraction method 2, and the extraction method 3, it operated similarly to Example 4 and calculated | required the viable cell count and the viable cell collection rate (yield). The results are shown in Table 2.
The chewing gum composition 3 produced in Example 3 was cut into a width of about 1 mm and a length of 15 mm to obtain a measurement sample.
About 1 g of this is accurately weighed and placed in a 20 ml test tube. 1) Was exactly 10 ml, and lactic acid bacteria were extracted by the following extraction method.
Extraction method 1: mixed for 1 minute with a test tube mixer and then allowed to stand for 5 minutes. This operation was repeated 5 times, and further kneaded with a spatula for 15 minutes to obtain an extract.
About the extract obtained by the extraction method 1, it operated similarly to Example 4 and calculated | required the number of viable bacteria. The results are shown in Table 3.
A chewing gum composition was prepared under the conditions shown below according to the production method of Example 1, using 7 g of gum base, 9 g of xylitol, 4 g of mannitol and 0.35 g of Lactobacillus salivarius bacteria triturate.
Chewing gum composition 7-1; kneading temperature 50 ° C., kneading time 1 hour
Chewing gum composition 7-2; kneading temperature 50 ° C., kneading time 3 hours
Chewing gum composition 7-3; kneading temperature 60 ° C., kneading time 1 hour
Chewing gum composition 7-4; kneading temperature 60 ° C., kneading time 3 hours
Chewing gum composition 7-5; kneading temperature 80 ° C., kneading time 1 hour
Chewing gum composition 7-6; kneading temperature 80 ° C., kneading time 3 hours
For each of these six chewing gum compositions, an extract was obtained according to Extraction Method 3 described in Example 5, and the viable cell count and recovery rate were determined. The results are shown in Table 4.
The chewing gum composition 2 obtained by the production method shown in Example 2 was cut into pieces of about 1 mm in width and 15 mm in length, accurately weighed about 1 g, put into a 20 ml test tube, and preheated to 37 ° C. Diluted solution 1) 10 ml of was added. The test tube was allowed to stand in a 37 ° C. water bath for 5 minutes and then stirred for 1 minute with a test tube mixer for 150 minutes. During this time, 0.5 ml of the extract was collected over time (30 minutes, 45 minutes, 60 minutes, 90 minutes, 120 minutes, 150 minutes after the start of the test) 1) Was diluted to obtain a test solution, and the number of viable bacteria was determined according to Example 4.
The chewing gum composition 3 obtained by the production method shown in Example 3 was also tested in the same manner. The results are shown in Table 5.
In this test, Streptococcus mutans (JCM5705 strain, hereinafter referred to as Sm) and Ls, GG, and La1 lactic acid strains were used. Moreover, the used culture medium is as follows.
Medium A: GAM medium supplemented with 1% glucose (Nissui Pharmaceutical)
Medium B: Medium A supplemented with xylitol to 1%
Viable lactic acid bacteria count medium: Modified LBS agar medium (Tomochika Mitsuoka, Intestinal Bacteriology, page 477, Asakura Shoten, 1990)
Sm viable cell count medium: GAM agar medium supplemented with bacitracin at 10 μg / ml and glucose at 1%
Lactic acid bacteria and Sm were precultured overnight at 37 ° C. in medium A to obtain each bacterial solution. 50 μl of this bacterial solution was inoculated into 50 ml of medium A and medium B, lactic acid bacteria and Sm were cultured alone and mixed at 37 ° C., and the culture solution was collected over time. The collected culture solution (50 μl) was appropriately diluted, applied to a lactic acid bacteria viable count medium and an Sm viable count assay medium, and anaerobically cultured at 37 ° C. for 3 days. The number of colonies produced was counted to determine the number of viable bacteria. The results are shown in Tables 6 to 8.
Test Example 2 Effect of a mixture of lactic acid bacteria, lactic acid bacteria and xylitol on the growth of Porphyromonas gingivalis
In this test, Porphyromonas gingivalis (JCM8525 strain, hereinafter referred to as Pg) and the lactic acid strain shown in Test Example 1 were used. Moreover, the used culture medium is as follows.
Medium A: GAM medium supplemented with 1% glucose (Nissui Pharmaceutical)
Medium B: Medium A supplemented with xylitol to 1%
Pg viable cell count medium: EG agar medium supplemented with gentamicin sulfate to a concentration of 10 μg / ml (Tomotsuka Mitsuoka, Intestinal Bacteriology, page 475, Asakura Shoten, 1990)
Lactic acid bacteria and Pg were precultured overnight at 37 ° C. in medium A to obtain each bacterial solution. 50 μl of this bacterial solution was inoculated into 50 ml of medium A and medium B, and single culture of Pg and mixed culture with lactic acid bacteria were performed at 37 ° C., and the culture solution was collected over time. 50 μl of the collected culture solution was appropriately diluted, applied to a Pg viable cell count medium, and anaerobically cultured at 37 ° C. for 3 days. The number of colonies produced was counted to determine the number of viable bacteria. The results are shown in Tables 9 to 11.
Example 9 Production of Chocolate Composition 1
40 g of cacao mass and 20 g of cocoa butter were mixed and dissolved while heating in a thermostatic bath at 50 ° C. for 10 minutes. After adding 30 g of xylitol and stirring for 1 minute, Lactobacillus salivarius triturate (1.3 × 10 6 per gram) 10 10 g (including individual bacteria) was added and mixed for another 1 minute. After dispensing into a petri dish and cooling to room temperature, the mixture was cooled and solidified at 5 ° C. for 3 hours to obtain a chocolate composition 1. Three lots were manufactured by the same manufacturing method.
Example 10 Viable count measurement of chocolate composition 1
The produced chocolate composition 1 was chopped into a width of about 1 mm and a length of 15 mm to obtain a measurement sample.
About 1 g is accurately weighed and placed in a 20 ml test tube. 1) Was accurately dissolved in a thermostat bath at 50 ° C. for 5 minutes while stirring with a test tube mixer. 1 ml of this was accurately measured, diluted with a diluent, and diluted exactly 10 times. This 10-fold dilution operation was further repeated 4 times, and the number of viable bacteria was determined by the method described in Test Example 1. Further, the viable cell recovery rate (yield) was determined from the obtained viable cell count and the theoretical amount of lactic acid bacteria at the time of addition. The results are shown in Table 12.
150 g of xylitol (manufactured by Towa Kasei), 54 g of resis (manufactured by Towa Kasei), 6 g of methylcellulose (manufactured by Shin-Etsu Chemical), 45 g of partially pregelatinized starch (manufactured by Asahi Kasei Kogyo), 30 g of polydextrose (manufactured by Wako Pure Chemical Industries), sucrose fatty acid ester ( 6 g of Mitsubishi Chemical Foods Co., Ltd. 6 g of castor wax (manufactured by Nippon Oil & Fats), 3 g of light anhydrous silicic acid (manufactured by Freund Sangyo), Lactobacillus salivarius bacteria triturate (including 1.3 x 1011 viable bacteria per gram) 15 g was mixed well in a plastic bag and then tableted using a tableting machine (6B-2, manufactured by Kikusui Seisakusho) to obtain a chewable tablet having a diameter of 15 mm and a weight of 1000 mg.
Example 12 Production of Sustained Release Oral Insert
Rolling fluidized bed granulation of 150 g of xylitol (manufactured by Towa Kasei), 54 g of powdered reduced maltose (manufactured by Towa Kasei), 60 g of dextrin (manufactured by Matsutani Chemical Industry), 15 g of gluten (manufactured by Wako Pure Chemical Industries), and 6 g of methylcellulose (manufactured by Shin-Etsu Chemical) An 80% ethanol solution was sprayed and granulated using a machine (Numeralizer NQ-Labo, manufactured by Fuji Powder). The granulated product was dried by ventilation at 60 ° C. for 16 hours and then sized with an 18 mesh sieve to obtain granules for tableting. Lactobacillus salivarius triturate (1.3 x 10 per gram) 11 15 g) and 15 g of castor wax (Nippon Yushi Co., Ltd.) were added and mixed well, and then tableted using a tableting machine (RT-F9, manufactured by Kikusui Seisakusho), with a sustained release of 6 mm in diameter and 100 mg in weight. Sex oral insertion tablets were obtained.
Test Example 3 Effect of Lactic Acid Bacteria, Mixture of Lactic Acid Bacteria and Xylitol on Production of Insoluble Glucan by Streptococcus mutans (Sm)
Ls was used for lactic acid bacteria. Moreover, the used culture medium is as follows.
Medium A: GAM medium supplemented with 2% sucrose (Nissui Pharmaceutical)
Medium B: Medium A supplemented with xylitol to 1%
Sm and Ls were each precultured overnight at 37 ° C. in medium A to obtain each bacterial solution. 0.1 ml of this bacterial solution was inoculated into 10 ml of medium A and medium B, and single culture and mixed culture of Sm and Ls were performed at 37 ° C. for 24 hours. The culture solution was centrifuged (8000 rpm, 20 minutes) to recover the precipitate (bacteria + insoluble glucan), and washed 3 times with 10 ml of phosphate buffered physiological solution (hereinafter referred to as PBS). The insoluble glucan was dissolved by adding 5 ml of 0.1N NaOH, centrifuged (8000 rpm, 20 minutes), and the supernatant was collected. The sugar concentration in the supernatant was measured by the phenol sulfate method, and the amount of insoluble glucan produced was examined. To 0.2 ml of the supernatant, 0.2 ml of 5% phenol and 1 ml of concentrated sulfuric acid were added, allowed to stand for 10 minutes, mixed, and further allowed to stand for 20 minutes. The absorbance at 492 nm of the reaction solution was measured, and the concentration of insoluble glucan was calculated based on a calibration curve prepared using a glucose solution. The results are shown in Table 13.
Test Example 4 Search for a substance that enhances the growth inhibitory effect of Porphyromonas gingivalis by lactic acid bacteria culture solution (1)
A search was made for a substance that enhances the growth inhibitory effect of Porphyromonas gingivalis (Pg) by lactic acid bacteria, such as xylitol. The lactic acid bacterium used is Ls, and the medium is as follows.
Medium A: GAM medium supplemented with 1% glucose (Nissui Pharmaceutical)
Medium B: Medium A with Ls culture filtrate added to 30%
The culture filtrate of Ls was prepared by centrifuging (8000 rpm, 20 minutes) the culture solution of Ls cultured at 37 ° C. for 24 hours in medium A, collecting the supernatant, and further performing sterile filtration.
Pg viable cell count medium: EG agar medium supplemented with gentamicin sulfate to a concentration of 10 μg / ml (Tomotsuka Mitsuoka, Intestinal Bacteriology, page 475, Asakura Shoten, 1990)
100 μl of the preculture medium in which Pg was cultured overnight at 37 ° C. in medium A was inoculated into medium B supplemented with 10 ml of medium B and 10% of various substances, and cultured at 37 ° C. for 24 hours. The culture solution was appropriately diluted, applied to a Pg viable count medium, and anaerobically cultured at 37 ° C. for 3 days. The number of colonies produced was counted to determine the number of viable bacteria. The results are shown in Table 14.
Test Example 5 Effects of Lactic Acid Bacteria Culture Solution and Xylitol on the Growth of Actinobacillus actinomycetemcomitans
Actinobacillus actinomycetemcomitans (JCM8577 strain, hereinafter referred to as Aa) was used. The lactic acid bacterium used in this test example is Ls, and the medium is as follows.
Medium A: Brain Heart Infusion liquid medium (Becton Dickinson)
Medium B: Medium A with Ls culture filtrate added to 25%
The Ls culture filtrate was obtained by centrifuging (8000 rpm, 20 minutes) the Ls culture solution cultured at 37 ° C. for 24 hours in a GAM medium supplemented with glucose to 1%, and collecting the supernatant. Prepared by filtration.
Aa Viable count medium: Brain Heart Infusion Agar (Becton Dickinson)
A precultured solution of Aa cultured overnight at 37 ° C. in medium A is about 10 viable cells. 6 10 ml of each medium (medium A, medium B, medium A supplemented with 3%, 7% xylitol and medium B supplemented with 3%, 7% xylitol) at 37 ° C. Microaerobic culture was performed for 48 hours. The culture solution was appropriately diluted, applied to the Aa viable cell count medium, and anaerobically cultured at 37 ° C. for 3 days. The number of colonies produced was counted to determine the number of viable bacteria. The results are shown in Table 15.
Test Example 6 Effect of lactic acid bacteria culture solution and xylitol on the growth of Fusobacterium nucleatum
Fusobacterium nucleatum (JCM8532T strain, hereinafter referred to as Fn) was used. The lactic acid bacterium used in this test example is Ls, and the medium is as follows.
Medium A: GAM medium supplemented with 1% glucose (Nissui Pharmaceutical)
Medium B: Medium A supplemented with 50% Ls culture filtrate
The Ls culture filtrate was prepared by the same method as in Test Example 5.
Viable count medium for Fn: Brain Heart Infusion Agar (Becton Dickinson)
About 10 viable cells were obtained by pre-cultured Fn in medium A at 37 ° C. overnight. 6 10 ml of each medium (medium A, medium B, medium A supplemented with 3%, 10% xylitol and medium B supplemented with 3%, 10% xylitol) at 37 ° C. Anaerobic culture was performed for 24 hours. The culture solution was appropriately diluted, applied to the Fn viable count medium, and anaerobically cultured at 37 ° C. for 3 days. The number of colonies produced was counted to determine the number of viable bacteria. The results are shown in Table 16.
Test Example 7 Inhibitory effect of lactic acid bacteria culture solution and xylitol on adhesion of Porphyromonas gingivalis to oral cells
Ls was used for lactic acid bacteria. The Ls culture filtrate was obtained by centrifuging (8000 rpm, 20 minutes) the Ls culture solution cultured at 37 ° C. for 24 hours in a GAM medium supplemented with glucose to 1%, and collecting the supernatant. After filtration, neutralized with 1N sodium hydroxide.
The cells were cultured overnight at 37 ° C. in a GAM medium supplemented with 1 g of Pg glucose. Wash with PBS and have 10 viable bacteria 7 The following media (medium A, medium B, medium A supplemented with 2.5%, 5%, 10% xylitol and 2.5%, 5%, 10% xylitol were added so as to be / ml. Suspended in medium B) to prepare a Pg suspension for cell attachment.
Medium A: DMEM / F-12 medium (Kibuko)
Medium B: Medium A with Ls culture filtrate added to 25%
As oral cells, human oral cultured cells HO1-u-1 (JCRB0828 strain) were used. Cells are 4x10 5 seeded in a 24-well plate to cells / well, 37 ° C, 5% CO 2 A monolayer was formed by culturing overnight under conditions. After washing 3 times with PBS, 1 ml of Pg suspension for cell attachment was added, and the mixture was incubated at 37 ° C., 5% CO2. 2 It was allowed to stand for 1.5 hours under the conditions. The cells were washed 3 times with PBS, 1 ml of sterilized water was added, and the cells were left at room temperature for 10 minutes to detach the cells. After appropriate dilution with PBS, the viable count of Pg adhering to the cells was measured. For the measurement, an EG agar medium containing 5% horse defibrinated blood was used and cultured at 37 ° C. under anaerobic conditions for 3 days. The results are shown in Table 17.
Test Example 8 Search for a substance that enhances the growth inhibitory effect of Porphyromonas gingivalis by lactic acid bacteria culture solution (2)
We searched for substances that enhance the growth-inhibiting effect of Porphyromonas gingivalis by lactic acid bacteria, such as xylitol. The lactic acid bacterium used in this test example is Ls, and the medium is as follows.
Medium A: GAM medium supplemented with 1% glucose (Nissui Pharmaceutical)
Medium B: Medium A with Ls culture filtrate added to 25%
The culture filtrate of Ls was prepared by centrifuging (8000 rpm, 20 minutes) the culture solution of Ls cultured at 37 ° C. for 24 hours in medium A, collecting the supernatant, and further performing sterile filtration.
Pg viable cell count medium: EG agar medium supplemented with gentamicin sulfate at 10 μg / ml
100 μl of the preculture medium in which Pg was cultured overnight at 37 ° C. in medium A was inoculated into 10 ml of medium B and medium B supplemented with 0.2 to 10% of various substances, and cultured at 37 ° C. for 9 hours. The culture solution was appropriately diluted, applied to a Pg viable count medium, and anaerobically cultured at 37 ° C. for 3 days. The number of colonies produced was counted to determine the number of viable bacteria. The results are shown in Table 18 and Table 19. In addition, since the experiment shown in Table 18 and Table 19 was performed independently, the numerical value of the Pg viable count in the case of no additive substance addition in Table 18 and the Pg viable count in the case of no additive addition in Table 19 are different. Yes.
Various plant extracts, red yeast rice, turmeric pigment, enzyme-decomposed licorice, and glycyrrhizin manufactured by Maruzen Pharmaceutical Co., Ltd. were used.
Test Example 9 Effects of Lactic Acid Bacteria (Bifidobacterium) Culture Solution and Xylitol on the Growth of Porphyromonas gingivalis
The lactic acid bacteria used in this test example were bifidobacteria longum (Bifidobacterium longum WB1001 strain (FERM P-12610), hereinafter referred to as Bl) and Bifidobacterium bifidum WB1003 strain (FER). -12612), hereinafter Bb), and the medium is as follows.
Medium A: GAM medium supplemented with 1% glucose (Nissui Pharmaceutical)
Medium B: Medium A supplemented with 20% of the culture filtrate of Bl
Medium C: Medium A with Bb culture filtrate added to 35%
The culture filtrates of B1 and Bb were prepared by centrifuging (8000 rpm, 20 minutes) the culture solution obtained by culturing each strain in medium A at 37 ° C. for 20 hours, collecting the supernatant, and further performing sterile filtration.
Pg viable cell count medium: EG agar medium supplemented with gentamicin sulfate at 10 μg / ml
Pg was cultured in medium A at 37 ° C. overnight. 100 μl of the preculture was inoculated into 10 ml of medium B, medium C, medium B supplemented with 10% xylitol and medium C supplemented with 10% xylitol. For 9 hours. The culture solution was appropriately diluted, applied to a Pg viable count medium, and anaerobically cultured at 37 ° C. for 3 days. The number of colonies produced was counted to determine the number of viable bacteria. The results are shown in Table 20.
Test Example 10 Effects of lactic acid bacteria culture solution and xylitol on the growth of Prevotella intermedia
Prevoterra Intermedia (JCM7365 strain, hereinafter referred to as Pi) was used. The lactic acid bacterium used in this test example is Ls, and the medium is as follows.
Medium A: GAM medium supplemented with 1% glucose (Nissui Pharmaceutical)
Medium B: Medium A supplemented with 50% Ls culture filtrate
The Ls culture filtrate was prepared by the same method as in Test Example 5.
Pi viable cell count medium: EG agar medium containing 5% equine defibrinated blood
10 ml of each medium (medium A, medium B, medium A supplemented with 10% xylitol and 10% of the pre-cultured solution in which Pi was cultured overnight at 37 ° C. in medium A to have a viable cell count of about 107 / ml The medium B) supplemented with xylitol was inoculated and anaerobically cultured at 37 ° C. for 24 hours. The culture solution was appropriately diluted, applied to a Pi viable cell count medium, and anaerobically cultured at 37 ° C. for 3 days. The number of colonies produced was counted to determine the number of viable bacteria. The results are shown in Table 21.
Industrial applicability
At least one selected from the group consisting of the living lactic acid bacterium of the present invention, the lactic acid bacterium-containing material, the lactic acid bacterium culture filtrate, and a processed product thereof, and xylitol, sorbitol, erythritol, trehalose, maltitol, mannitol, palatinose, reduction Lactose, sucrose, fructose, glucose, fructooligosaccharides, aspartame, sorghum extract, dokudami extract, red yeast rice, turmeric pigment, karin extract, ganoderma extract, hawthorn extract, shiitake extract, moon peach leaf extract, Stevia extract, Lotus germ extract, Lacanca extract, Green tea extract, Maria thistle extract, Purple brown rice extract, Enzyme-decomposed licorice, Twilight leaf extract, Ginkgo biloba extract, Rooibos tea extract, Artemisia extract, Glycyrrhizin, Containing at least one selected from the group consisting of a gymnema extract and a prickly pear extract Compositions for prevention and / or treatment of intraluminal diseases, especially chewing gum compositions, can retain lactic acid bacteria continuously in the oral cavity due to good sustained release properties, and are safe, simple and efficient in the oral cavity Can be used for disease prevention and treatment.
Claims (11)
乳酸菌が、ラクトバチルス・サリバリウス、ラクトバチルス・アシドフィルス、ラクトバチルス・ラムノーサス及びラクトバチルス・ジョンソニーからなる群から選ばれる１種又は２種以上である上記組成物。 At least one selected from the group consisting of a living lactic acid bacterium, the lactic acid bacterium-containing product, the lactic acid bacterium culture filtrate, and a processed product thereof, and xylitol, sorbitol, erythritol, trehalose, maltitol, mannitol, palatinose, reduced lactose, Sugar, fructose, dextrose, fructooligosaccharide, aspartame, sorghum extract, wolfberry extract, red yeast rice, turmeric pigment, karin extract, ganoderma extract, hawthorn extract, shiitake extract, moon peach leaf extract, stevia extract , Lotus germ extract, Lacanca extract, Green tea extract, Maria thistle extract, Purple brown rice extract, Enzymatic degradation licorice, Yellow leaves extract, Ginkgo biloba extract, Rooibos tea extract, Artemisia extract, Glycyrrhizin, Gymnema extract , And at least one selected from the group consisting of prickly pear extracts or A prophylactic and / or therapeutic composition of the disease,
The said composition whose lactic acid bacteria are 1 type (s) or 2 or more types chosen from the group which consists of Lactobacillus salivaius, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus johnsonii.
乳酸菌が、ラクトバチルス・サリバリウス、ラクトバチルス・アシドフィルス、ラクトバチルス・ラムノーサス及びラクトバチルス・ジョンソニーからなる群から選ばれる１種又は２種以上である上記組成物。 At least one selected from the group consisting of a living lactic acid bacterium, the lactic acid bacterium-containing product, the lactic acid bacterium culture filtrate, and a processed product thereof, and xylitol, sorbitol, erythritol, trehalose, maltitol, mannitol, palatinose, reduced lactose, Sugar, fructose, dextrose, fructooligosaccharide, aspartame, sorghum extract, wolfberry extract, red yeast rice, turmeric pigment, karin extract, ganoderma extract, hawthorn extract, shiitake extract, moon peach leaf extract, stevia extract , Lotus germ extract, Lacanca extract, Green tea extract, Maria thistle extract, Purple brown rice extract, Enzymatic degradation licorice, Yellow leaves extract, Ginkgo biloba extract, Rooibos tea extract, Artemisia extract, Glycyrrhizin, Gymnema extract , And at least one selected from the group consisting of prickly pear extracts and / Or a cytostatic composition for periodontal bacteria,
The said composition whose lactic acid bacteria are 1 type (s) or 2 or more types chosen from the group which consists of Lactobacillus salivaius, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus johnsonii.
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| JP2001102752 | 2001-04-02 | ||
| JP2001102752 | 2001-04-02 | ||
| JP2001267943 | 2001-09-04 | ||
| JP2001267943 | 2001-09-04 | ||
| PCT/JP2002/003293 WO2002080946A1 (en) | 2001-04-02 | 2002-04-02 | Compositions for preventing and/or treating oral diseases |
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| JPWO2002080946A1 JPWO2002080946A1 (en) | 2004-07-29 |
| JP3921175B2 true JP3921175B2 (en) | 2007-05-30 |
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| JP2002578985A Expired - Lifetime JP3921175B2 (en) | 2001-04-02 | 2002-04-02 | Composition for prevention and / or treatment of oral disease |
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| Country | Link |
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| JP (1) | JP3921175B2 (en) |
| AU (1) | AU2002243023A1 (en) |
| WO (1) | WO2002080946A1 (en) |
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| KR20110019420A (en) * | 2008-06-11 | 2011-02-25 | 바스프 에스이 | Use and methods for preventing and/or treating oral malodour |
| KR20200121173A (en) * | 2019-04-15 | 2020-10-23 | 주식회사 오라팜 | Composition for preventing or improving periodontal disease comprising green tea extract and lactic acid bacteria, cultured products, fragmented products or extract of the same as an effective ingredient |
| KR102488050B1 (en) * | 2021-10-29 | 2023-01-13 | 주식회사 쎌바이오텍 | Antibacterial composition comprising lactobacillus strain culture filtrate |
Families Citing this family (26)
| Publication number | Priority date | Publication date | Assignee | Title |
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Family Cites Families (22)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IL48997A (en) * | 1975-02-20 | 1978-10-31 | Sparamedica Ag | Therapeutic compositions containing xylitol for treating dental caries |
| CA1262442A (en) * | 1984-03-09 | 1989-10-24 | Yasuo Kawai | Anticariogenic or antiperiodontitic agent |
| US4912089A (en) * | 1986-11-07 | 1990-03-27 | Yasutake Hiji | Cariostatic materials and foods, and method for preventing dental caries |
| JP2667422B2 (en) * | 1988-01-26 | 1997-10-27 | 株式会社伊藤園 | Gargle |
| JP2793271B2 (en) * | 1989-07-21 | 1998-09-03 | 日本製紙株式会社 | Low calorie low carious candy with improved palatability and method for producing the same |
| JPH045222A (en) * | 1990-04-20 | 1992-01-09 | Kao Corp | Crude drug extract for oral cavity and composition for oral cavity containing the same as active ingredient |
| JPH06179780A (en) * | 1992-12-15 | 1994-06-28 | Mitsubishi Petrochem Co Ltd | Polyolefin composition |
| FR2715582B1 (en) * | 1994-02-02 | 1996-03-15 | Centre Nat Rech Scient | Microcapsules with crosslinked flavonoid walls and compositions containing them. |
| JP3281716B2 (en) * | 1994-03-24 | 2002-05-13 | 丸善製薬株式会社 | Glucosyltransferase inhibitors, oral preparations and foods and drinks |
| JPH08310931A (en) * | 1995-05-19 | 1996-11-26 | Maruzen Pharmaceut Co Ltd | Inhibitor of glucosyltransferase and cleaning agent for oral cavity |
| JP3499969B2 (en) * | 1995-06-23 | 2004-02-23 | 明治製菓株式会社 | Sugar sweetener containing crystalline kestose |
| JP3590689B2 (en) * | 1996-03-07 | 2004-11-17 | 東和化成工業株式会社 | Composition for confectionery with low caries induction and method for producing confectionery using the same |
| JPH1081617A (en) * | 1996-07-15 | 1998-03-31 | Lion Corp | Dentifrice composition for prevention of dental caries |
| GB2317339A (en) * | 1996-08-01 | 1998-03-25 | Procter & Gamble | Dentifrice composition containing a curcuminoid |
| JPH10139646A (en) * | 1996-11-13 | 1998-05-26 | Maruzen Pharmaceut Co Ltd | Glucosyltransferase inhibitor, oral cavity composition and food and beverage |
| KR100266752B1 (en) * | 1997-08-07 | 2001-03-02 | 오종석 | Plaque inhitition by novel human oral strains of lacticacid producing bacteria |
| JP3648587B2 (en) * | 1998-03-04 | 2005-05-18 | サンスター株式会社 | Periodontal disease prevention or periodontal disease progression food composition |
| JP3763075B2 (en) * | 1998-04-24 | 2006-04-05 | サンスター株式会社 | Food composition, oral composition and pharmaceutical composition for prevention or treatment of periodontal disease |
| JP2000072790A (en) * | 1998-09-01 | 2000-03-07 | Maruzen Pharmaceut Co Ltd | Method of removing residual agrochemical in plant extract |
| JP2000154127A (en) * | 1998-11-18 | 2000-06-06 | Lion Corp | Composition for oral cavity |
| JP3650705B2 (en) * | 1999-03-24 | 2005-05-25 | サンスター株式会社 | Food containing palatinit |
| JP2001081021A (en) * | 1999-09-09 | 2001-03-27 | Ichimaru Pharcos Co Ltd | Cosmetic composition |
-
2002
- 2002-04-02 AU AU2002243023A patent/AU2002243023A1/en not_active Abandoned
- 2002-04-02 JP JP2002578985A patent/JP3921175B2/en not_active Expired - Lifetime
- 2002-04-02 WO PCT/JP2002/003293 patent/WO2002080946A1/en active Application Filing
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| KR20110019420A (en) * | 2008-06-11 | 2011-02-25 | 바스프 에스이 | Use and methods for preventing and/or treating oral malodour |
| KR101648992B1 (en) | 2008-06-11 | 2016-08-17 | 바스프 에스이 | Use and methods for preventing and/or treating oral malodour |
| KR20200121173A (en) * | 2019-04-15 | 2020-10-23 | 주식회사 오라팜 | Composition for preventing or improving periodontal disease comprising green tea extract and lactic acid bacteria, cultured products, fragmented products or extract of the same as an effective ingredient |
| KR102208751B1 (en) | 2019-04-15 | 2021-01-28 | 주식회사 오라팜 | Composition for preventing or improving periodontal disease comprising green tea extract and lactic acid bacteria, cultured products, fragmented products or extract of the same as an effective ingredient |
| KR102488050B1 (en) * | 2021-10-29 | 2023-01-13 | 주식회사 쎌바이오텍 | Antibacterial composition comprising lactobacillus strain culture filtrate |
| WO2023075290A1 (en) * | 2021-10-29 | 2023-05-04 | 주식회사 쎌바이오텍 | Antibacterial composition containing lactobacillus strain culture filtrate |
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| Publication number | Publication date |
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| WO2002080946A1 (en) | 2002-10-17 |
| JPWO2002080946A1 (en) | 2004-07-29 |
| AU2002243023A1 (en) | 2002-10-21 |
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