JP4571783B2 - Antimicrobial agent - Google Patents

Description

（式中、ｎは０以上の整数を表し、Ｒは水素原子又は炭化水素基を示す）
【００１１】
【化２】
化学式２：

（式中、ｎは０以上の整数を表し、Ｒは水素原子又は炭化水素基を示す）
【００１２】
【化３】
化学式３：

（式中、ｍ及びｎは０以上の整数を表し、Ｒは水素原子又は炭化水素基を示す）
【００１３】
【化４】
化学式４：

（式中、ｎは０以上の整数を表し、Ｒは水素原子又は炭化水素基を示す）
【００１４】
【化５】
化学式５：

（式中、ｎは０以上の整数を表し、Ｒは水素原子又は炭化水素基を示す）
【００１５】
本発明の抗微生物剤には、没食子酸の糖転移物及びカフェ酸の糖転移物の誘導体を用いることができる。なお、没食子酸及びカフェ酸への糖転移化と誘導体化の順序は問わない。没食子酸の糖転移物及びカフェ酸の糖転移物の誘導体としては、酸、アルカリ、加熱などの処理により、比較的容易に加水分解されて、没食子酸又はカフェ酸が生成され得る化合物が用いられ、それらは天然物であっても人工的な合成物であってもよい。例えば、天然に存在する没食子酸の誘導体としては、加水分解性タンニンと総称される、グルコース１分子に１乃至５分子の没食子酸がエステル結合した化合物、例えば、ガロイルグルコース、ケブリン酸、ケブラグ酸、コリラギンなどが、一方、カフェ酸の誘導体としてクロロゲン酸などが例示できる。これらの誘導体の糖転移物は、加水分解後に没食子酸の糖転移物又はカフェ酸の糖転移物を生成し、抗微生物効果を示すので、本発明の抗微生物剤の成分として使用することができる。没食子酸又はカフェ酸の誘導体に対して糖転移を行う際には、没食子酸及びカフェ酸部分に糖転移可能な水酸基を有する化合物であるのが好ましく、さらに、没食子酸又はカフェ酸部分以外に糖転移可能な水酸基を有しない化合物が好適である。そのような誘導体としては、例えば、化学式６に示すガロイルグルコースの一種であるペンタガロイルグルコース（１，２，３，４，６−ｐｅｎｔａｋｉｓ−Ｏ−ｇａｌｌｏｙｌ−β−Ｄ−ｇｌｕｃｏｓｅ）が挙げられる。
【００１６】
【化６】
化学式６：

【００１７】
さらに、没食子酸及びカフェ酸の誘導体としては、没食子酸及びカフェ酸の抗微生物効果を向上させるために、適宜の修飾を施した誘導体を利用することができる。このような誘導体としては、例えば、没食子酸及びカフェ酸のカルボキシル基に炭化水素基がエステル結合した化合物が挙げられる。エステル結合させる炭化水素基としては、鎖式炭化水素基であっても環式炭化水素基であってもよく、また、飽和炭化水素基であっても不飽和炭化水素基であってもよい。本発明における好適な炭化水素基は、鎖式炭化水素基、いわゆる脂肪族炭化水素基である。没食子酸及びカフェ酸と脂肪族炭化水素基とのエステル誘導体（以下、単にエステル誘導体ということもある。）は、脂肪族炭化水素基の炭素数に応じて抗微生物性が向上するという特徴を有しており、本発明の抗微生物剤として有利に用いられる。なお、没食子酸又はカフェ酸と脂肪族炭化水素基とのエステル誘導体は、炭化水素基の炭素数が多くなるほど水溶性に乏しくなる傾向があるものの、糖転移することにより水溶性が改善される。炭化水素基の炭素数は、本発明の抗微生物剤の使用方法、形態、対象となる微生物などに応じて適宜選択すればよく、通常、炭素数１乃至１２の範囲、好ましくは３乃至８の範囲の炭化水素基とのエステル誘導体に糖転移したものが本発明の抗微生物剤として用いられる。炭素数１２を超える炭化水素基とのエステル誘導体は、その水溶性が著しく低下しているので、糖転移によっても十分に水溶性が改善されず、本発明の抗微生物剤を生体に適用する際に好ましくない場合がある。また、没食子酸と炭素数３の脂肪族炭化水素基とのエステル誘導体である没食子酸ｎ−プロピルは、炭化水素基の炭素数が少ない割に抗微生物性が高く、その糖転移物は、本発明の抗微生物剤として特に有利に用いられる。
【００１８】
没食子酸の糖転移物、カフェ酸の糖転移物、及び、それらの誘導体（以下、単に「没食子酸の糖転移物」又は「カフェ酸の糖転移物」ということもある。）を得る方法としては、特に限定されず、化学合成法又は酵素反応法などによって製造可能である。このうち、糖転移酵素を利用した酵素反応法が簡便であることから、本発明においては有利に利用できる。例えば、没食子酸又はその誘導体、又は、カフェ酸又はその誘導体にデキストリン、サイクロデキストリン、澱粉あるいはこれらの混合物を添加し、これを、例えば、バチラス属に属する微生物が産生する糖転移酵素、例えば、バチラス・ステアロサーモフィラス由来のサイクロデキストリングルカノトランスフェラーゼ（ＥＣ ２．４．１．１９）を作用させて糖転移し、次いで所望により、この酵素反応液にグルコアミラーゼ及び／又はβ−アミラーゼを作用させる方法が挙げられる。この方法によれば、没食子酸の糖転移物及びカフェ酸の糖転移物の糖質部分の重合度を調節することが可能である。
【００１９】
また、没食子酸の糖転移物又はカフェ酸の糖転移物を得る方法として、これらを含有する植物体の抽出物をそのまま、又は精製工程の適宜の段階で、糖転移することも可能である。この場合、没食子酸又はその誘導体、又は、カフェ酸又はその誘導体とともに、他の成分の中に糖転移物となるものがあり、それらの成分の水溶性をも向上させるので、抽出物の水溶性を大幅に改善する効果も期待できる。没食子酸又はその誘導体、又は、カフェ酸又はその誘導体を含有する植物としては、緑茶、紅茶、コーヒーなどが一般的であり、その他に、モモタマナ属に属する植物、タデ藍（ポリゴナム属）などが挙げられるが、これらに限定されない。
【００２０】
本発明の抗微生物剤には、没食子酸の糖転移物又はカフェ酸糖転移物以外にも、製法に由来する未糖転移物である没食子酸又はその誘導体、又は、カフェ酸又はその誘導体（以下、単に「未糖転移物」ということもある。）が含まれることがある。未糖転移物は、高速液体クロマトグラフィー（ＨＰＬＣ）などの手法により糖転移物標品から除去可能であるが、未糖転移物を含んだまま本発明の抗微生物剤に配合することも、また必要に応じて、糖転移物とともに、それに対応する未糖転移物を本発明の抗微生物剤に配合することもできる。この場合、糖転移物の含有量は、糖転移物と未糖転移物との総和に対して、モル比で、通常１０％以上、好ましくは２０％以上、さらに好ましくは５０％以上とする。糖転移物の割合が１０％に満たない場合は、生体内での持続性及び水溶性を改善する効果が十分期待できず、好ましくない場合がある。
【００２１】
また、本発明で用いる没食子酸の糖転移物又はカフェ酸の糖転移物において、グルコース以外にも、ガラクトース、マンノース、フコースなどの単糖、グルコサミン、ガラクトサミンなどのアミノ糖なども糖転移させる糖として選択することができる。糖転移物における糖質部分の重合度は、好ましくは1乃至５、さらに好ましくは1乃至３から選択され、最も好ましくは１である。糖質部分の重合度の高い糖転移物ほど、没食子酸及びカフェ酸の生体内での持続性及び水溶性を改善する効果に優れ、遅効性の抗微生物剤とする効果が期待できるものの、即効性が低下する傾向にある。したがって、即効性を期待する場合には、重合度が５を超える糖質部分を有する糖転移物は好ましくない場合がある。
【００２２】
本発明の抗微生物剤は、没食子酸の糖転移物及び／又はカフェ酸の糖転移物を有効成分として含有し、さらには所望により没食子酸又はその誘導体、及び／又は、カフェ酸又はその誘導体を配合することできる。また、通常、注射用剤、皮膚外用剤、経粘皮用剤、経鼻腔用剤又は経口用剤を構成するための成分の１又は複数を配合することができる。
【００２３】
本発明の抗微生物剤は、用途に応じて、例えば、液状、ペースト状又は固状の注射用剤、皮膚外用剤、経粘皮用剤、経鼻腔用剤、経口用剤などの形態に調製して用いられ、具体的には、注射剤、点眼剤、座剤、軟膏剤、さらには、歯磨き、うがい薬などの口腔清浄剤や、歯科領域における口腔殺菌剤をはじめとする口腔用剤の形態が好ましい。したがって、本発明の抗微生物剤は、散剤、細粒剤、丸剤、錠剤、カプセル剤、トローチ剤、リモナーデ剤、エリキシル剤、シロップ剤、芳香水剤、懸濁剤、乳剤、浸剤、煎剤、チンキ剤、エキス剤、流エキス剤、酒精剤、リニメント剤、ローション剤、軟膏剤、パップ剤、硬膏剤、坐剤、点眼剤、眼軟膏剤、注射剤において汎用される、例えば、研磨剤、発泡剤、湿潤剤、粘結剤、香味料、甘味料、保存料、弗素化合物、酵素剤、消臭剤、さらには、賦形剤、軟膏基剤、溶解剤、矯味剤、矯臭剤、着色剤、乳化剤、噴射剤などの調製用薬などの１又は複数と組み合わせて用いることを妨げない。また、本発明の目的を逸脱しない範囲で、斯かる剤形の薬剤において汎用される、例えば、止血剤、抗炎症剤、組織賦活剤、抗微生物剤、さらには、抗生物質、インターフェロンα、インターフェロンβ、インターフェロンγをはじめとする１又は複数の他の薬効成分との併用も妨げない。なお、注射用剤や経粘皮用剤、経鼻腔、口腔用剤における調製用薬としては、没食子の酸糖転移物及びカフェ酸の糖転移物を安定化する性質を兼備する点で、例えば、スクロース、グルコース、マルトース、トレハロース、フルクトース、環状四糖、マンニトール、マルチトールなどの糖又は糖アルコールさらには、アラニン、アルギニン、アスパラギン、アスパラギン酸、シトルリン、システイン、シスチン、グルタミン、グリシン、ヒスチジン、イソロイシン、ロイシン、リジン、メチオニン、オルニチン、フェニルアラニン、フェニルグリシン、プロリン、セリン、スレオニン、トリプトファン、チロシン、バリン、ヒドロキシプロリン、γ−カルボキシグルタマート、Ｏ−ホスホセリン、β−アラニン、α−アミノ酪酸、γ−アミノ酪酸、α−アミノイソ酪酸、４−（４−アミノフェニル）酪酸、アミノフェニル酪酸、アミノ安息香酸、４−アミノ馬尿酸、アミノメチル安息香酸、ε−アミノカプロン酸、７−アミノへプタン酸、β−アスパラギン酸、γ−グルタミン酸、ε−リジン、メチオニンスルホン、ノルロイシン、ノルバリン、オルニチン、ｄ−オルニチン、ｐ−ニトロ−フェニルアラニン、ヒドロキシプロリン、チオプロリンなどのアミノ酸又はこれらのアミノ酸の誘導体若しくはオリゴペプチドが好ましく、必要に応じて、これらは組み合わせて用いられる。
【００２４】
本発明の抗微生物剤の抗微生物スペクトルは比較的広く、グラム陰性菌、グラム陽性菌、真菌、酵母などの多くの微生物に対して抗微生物効果を示す。本発明の抗微生物剤が対象とする微生物としては、例えば、バチラス属、ミコバクテリウム属、シュードモナス属、エッセリシア属、スタフィロコッカス属、ストレプトコッカス属、サルモネラ属、クレブシラ属、プロテウス属、ビブリオ属、シゲラ属、ヘリコバクター属、クラミジア属、アスペルギルス属、カンディダ属などに属する微生物が挙げられる。
【００２５】
本発明の抗微生物剤へ配合する没食子酸の糖転移物及び／又はカフェ酸の糖転移物の量は、その剤形、使用方法などを考慮して適宜決定すればよいが、通常、没食子酸又はカフェ酸換算で、没食子酸とカフェ酸の合計量が、０．００１乃至１００％（ｗ／ｗ）、好ましくは、０．０１乃至１００％（ｗ／ｗ）、さらに好ましくは０．１乃至１００％（ｗ／ｗ）の範囲内から選ばれる。また、投与量としては、抗微生物剤の適用対象、投与経路などにもよるけれども、通常、没食子酸又はカフェ酸換算で、没食子酸とカフェ酸の合計量が、１ｎｇ乃至１ｇ／日／成人、好ましくは、１０ｎｇ乃至１００ｍｇ／日／成人とする。
【００２６】
本発明の抗微生物剤は、斯かる用量で１日又は１週間に１回以上の頻度で注射又は経皮、経粘皮、経鼻腔用剤若しくは経口経路で適用することによって、生体における多種の病原性微生物の増殖を著明に抑制することから、鼻腔、口腔や気道、さらには視覚器、消化器、泌尿器、生殖器などへ侵入した病原性微生物が発症原因となる、例えば、肺炎などの呼吸器系疾患や、動脈硬化、虚血性心疾患などの循環器系疾患、結膜炎などの視覚器系疾患、直腸炎などの消化器系疾患、尿道炎などの泌尿器系疾患、副睾丸炎、卵管炎、子宮頚管炎、切迫早産、卵管の炎症、癒着、閉塞などの生殖器系疾患、食中毒の治療、予防に著効を発揮する。とりわけ、本発明の抗微生物剤は、抗生物質を有効成分とする経口用剤とは違って、耐性菌の出現を招来したり、長期連用しても大腸における有用な菌叢を破壊することがないので、食欲不振、嘔吐、下痢、便秘、膨満感を生じたり、倦怠感、発湿などの副作用を惹起することがない。
【００２７】
以上のとおり、没食子酸の糖転移物及び／又はカフェ酸の糖転移物は、生体中での持続性及び水溶性に優れるという利点を兼ね備えている。これらの特徴は、炭素数３以上の脂肪族炭化水素基を有する没食子酸エステル誘導体、カフェ酸及びカフェ酸エステル誘導体のように、水溶性に乏しい化合物を糖転移した場合にも発揮される。また、没食子酸の近接する３つの水酸基は鉄イオンと錯体を形成し、変色することが知られているところ、糖転移することによって、没食子酸の変色を防止する効果も期待できる。
【００２８】
以下、本発明による抗微生物剤の性状及び効果につき、実験例に基づいて説明する。
【００２９】
【実験例１】
＜没食子酸、没食子酸エステル誘導体及びカフェ酸の糖転移物の調製＞
１質量部の没食子酸、没食子酸メチル、没食子酸エチル、没食子酸ｎ−プロピル、没食子酸オクチル、没食子酸ドデシル（以上、関東化学株式会社製）又はカフェ酸（和光純薬工業株式会社製）を、それぞれ１０質量部の澱粉部分分解物（ＤＥ８、商品名『パインデックス＃１』、松谷化学株式会社製）とともに２００質量部の水に添加して加熱溶解した。適量の水酸化ナトリウムを添加してｐＨ６．０に調整した後、澱粉部分分解物１ｇあたり２００Ｕのサイクロデキストリングルカノトランスフェラーゼ（株式会社林原生物化学研究所製）を加え、４０℃で２４時間インキュベートした。その後、１００℃で２０分間インキュベートして酵素を失活させた後、適量の水酸化ナトリウムを添加してｐＨ４．５に調整した後、澱粉部分分解物１ｇあたり５０Ｕのグルコアミラーゼ（商品名『グルコチーム』、ナガセ生化学工業株式会社製）を加え、４０℃で２０時間インキュベートした。次いで、８０℃で１０分間インキュベートして酵素を失活させた後、ろ紙でろ過した。それぞれの反応液に、没食子酸、没食子酸メチル、没食子酸エチル、没食子酸ｎ−プロピル、没食子酸オクチル、没食子酸ドデシル、又はカフェ酸１ｇあたり２００ｍｌの合成吸着剤（商品名『ＨＰ１０』、三菱化学株式会社製）を充填したカラムに負荷して吸着させ、水で洗浄後、５％（ｗ／ｗ）エタノール水溶液でさらに洗浄し、各試料の疎水性に応じて、１５乃至５０％（ｗ／ｗ）エタノール水溶液で溶出した。これらを、減圧乾固した後、水に再溶解した後、同じカラムで同じ操作を繰り返し、溶出液を減圧乾固して、本発明の抗微生物剤に用いる没食子酸の糖転移物、没食子酸メチルの糖転移物、没食子酸エチルの糖転移物、没食子酸ｎ−プロピルの糖転移物、没食子酸オクチルの糖転移物、没食子酸ドデシルの糖転移物、カフェ酸の糖転移物を得た。これらを常法の高速液体クロマトグラフィー（ＨＰＬＣ）分析により、紫外部吸収を測定し組成比（モル比）を求めたところ、調製された糖転移物はすべてがモノグルコシドであり、それらの純度はどれもが９５％以上であった。
なお、サイクロデキストリングルカノトランスフェラーゼの１Ｕは、１ｍＭ塩化カルシウムを含む２０ｍＭ酢酸緩衝液（ｐＨ５．５）に加熱溶解した０．３％（ｗ／ｗ）可溶性澱粉粉溶液５ｍｌに酵素液０．２ｍｌを加え、４０℃、１０分間反応させ、この反応液０．５ｍｌを０．０２Ｎの硫酸１５ｍｌに加え反応を停止し、これに、０．１Ｎヨウ素溶液を０．２ｍｌを加えた後、６６０ｎｍの吸光度を測定する場合において、吸光度を１０％減少させる酵素量と定義されている。また、グルコアミラーゼの１Ｕは、１．０％（ｗ／ｗ）可溶性澱粉粉溶液（２０ｍＭ酢酸緩衝液、ｐＨ４．５）５ｍｌに、酵素液０．２ｍｌを加え、４０℃、１０分間反応させ、生成した還元糖量をソモギー−ネルソン法で測定する場合において、４０℃、１分間に１μｍｏｌのグルコースに相当する還元力を遊離する酵素量と定義されている。
【００３０】
【実験例２】
＜没食子酸、没食子酸エステル誘導体及びカフェ酸の糖転移物の水溶性＞
実験例１で得られた糖転移物の３７℃の水に対する溶解度を測定し、それぞれの未糖転移物と比較した。結果を表１に示す。
【００３１】
【表１】

【００３２】
表１に示す結果から、没食子酸、没食子酸エステル誘導体及びカフェ酸の糖転移物は、それぞれの未糖転移物よりも水溶性に優れており、特に、比較的水溶性に乏しい没食子酸ｎ−プロピル、没食子酸オクチル、没食子酸ドデシル及びカフェ酸については、水溶性の改善効果が顕著であった。また、別途、転移した糖の重合度が１を超える糖転移物について、同様の実験を行ったところ、糖転移された糖質部分の重合度が高いものほど水溶性を改善する効果に優れていた。
【００３３】
【実験例３】
＜各種糖転移物の抗微生物効果＞
実験例１により調製した各種糖転移物、後述の実施例５で調製したターミナリア ベリリカ抽出物の糖転移物、及び、後述の実施例６で調製したタデ藍抽出物の糖転移物の抗微生物効果を、表２に記載の微生物について、常法にしたがい、最小生育阻止濃度（ＭＩＣ）を測定した。また、対照として、それぞれの未糖転移物を用いた。結果を表２に示す。なお、表２における没食子酸メチル、没食子酸エチル、没食子酸ｎ−プロピル、没食子酸オクチル、没食子酸ドデシル及びその糖転移物のグラム数は、没食子酸換算での数値を示し、ターミナリア ベリリカ抽出物及びタデ藍抽出物、及びそれらの糖転移物は固形分のグラム数で示した。
【００３４】
【表２】

【００３５】
表２の結果から、微生物に対する最小生育阻止濃度は、糖転移の結果、同等乃至１／３２の範囲で低下するものの、抗微生物効果は保持されることが判明した。一方、ターミナリア ベリリカ抽出物及び藍抽出物の場合、有効成分としての没食子酸又はカフェ酸の糖転移物の含量が抽出物の固形分あたり約１％であることを考慮すると抗微生物効果が意外に優れており、他の成分による効果が加算されていることが想定された。また、糖転移物の糖質部分の重合度が高いもので同様の試験を行ったところ、重合度が高くなるほど、抗微生物活性が低下する傾向が認められた。
【００３６】
【実験例４】
＜生体内での持続性＞
実験例１で調製した各種糖転移物を、没食子酸又はカフェ酸換算で、０．１ｍｇ／ｍｌの水溶液に調製し、除菌ろ過した。一方、後述の実施例５で調製したターミナリア ベリリカ抽出物の糖転移物及び後述の実施例６で調製したタデ藍抽出物の糖転移物を、それが含有する没食子酸又はカフェ酸が０．１ｍｇ／ｍｌの水溶液となるように調製し、除菌ろ過した。これらをそれぞれ５週齢の雌性ウイスターラットの大腿部静脈に１ｍｌ（０．１ｍｇ）注射した。一定時間放置した後、ラットの尾静脈から経時的に血液を採取し、血清中の没食子酸、カフェ酸の含量を常法の液体高速クロマトグラフィー法により測定した。また、対照として、それぞれの未糖転移物を用いて同様に試験した。その結果、未糖転移物は静脈注射後約２時間で血中から検出できなくなるのに対し、糖転移物の場合は静脈注射後８時間までは血中から没食子酸、没食子酸エステル誘導体又はカフェ酸を検出できた。この結果は、糖転移物は、未糖転移物よりも生体内での持続性に優れることを物語っている。また、同時に、糖質部分の重合度が高い糖転移物で同様の試験を行ったところ、重合度が高くなるにつれて没食子酸、没食子酸エステル誘導体及びカフェ酸の生体内での持続性が向上する傾向が認められた。
【００３７】
【実験例５】
＜急性毒性試験＞
実験例１で調製した各種糖転移物を、常法にしたがって８週齢の雄性マウス各１０匹に経皮、経口あるいは腹腔内に投与した。その結果、これらのＬＤ_５０は、いずれの投与経路による場合にも約１００ｍｇ／ｋｇ（マウス体重）以上であった。この結果は、没食子酸の糖転移物及びカフェ酸の糖転移物がヒトへの投与を前提とする医薬品として安全であることを物語っている。
【００３８】
以上の結果から、没食子酸の糖転移物及び／又はカフェ酸の糖転移物は、これらの未糖転移物よりも抗微生物効果に劣るものの、抗微生物剤として有用なレベル以上の抗微生物効果を有しており、さらに、生体内での持続性及び水溶性に優れていた。これらの糖転移物が有する特徴は、本発明の抗微生物剤が、生体に適用する抗微生物剤として有利であることを示している。
【００３９】
以下、本発明の実施の形態につき、実施例に基づいて説明する。
【００４０】
【実施例１】
＜没食子酸糖転移物の調製＞
【００４１】
【実施例１−１】
１質量部の没食子酸（関東化学株式会社製）を、１０質量部の澱粉部分分解物（ＤＥ８、商品名『パインデックス＃１』、松谷化学株式会社製）とともに２００質量部の水に添加して加熱溶解した。適量の水酸化ナトリウムを添加してｐＨ６．０に調整した後、澱粉部分分解物１ｇあたり２００Ｕのサイクロデキストリングルカノトランスフェラーゼ（株式会社林原生物化学研究所製）を加え、４０℃で２４時間インキュベートした。その後、１００℃で２０分間インキュベートして酵素を失活させた後、ろ紙でろ過して、さらに、合成吸着剤（商品名『ＨＰ１０』、三菱化学株式会社製）に負荷して吸着させ、水で洗浄して澱粉部分分解物を溶出させた後、１０乃至２０％（ｗ／ｗ）エタノール水溶液で溶出した画分を回収し、減圧乾固し、本発明の抗微生物剤に用いる没食子酸糖転移物を得た。
【００４２】
これを常法の高速液体クロマトグラフィー（ＨＰＬＣ）分析に供し、紫外部吸収を測定し組成比（モル比）を求めたところ、グルコース重合度が１の糖質部分を有する没食子酸糖転移物が１０％、グルコース重合度が２の糖質部分を有する没食子酸糖転移物が４％、グルコース重合度が６以上の糖質を有する没食子酸糖転移物が６％、未反応のまま残った没食子酸酸が８０％であった。
【００４３】
【実施例１−２】
実施例１−１で得られたサイクロデキストリングルカノトランスフェラーゼ処理後の反応液に、適量の水酸化ナトリウムを添加してｐＨ４．５に調整した後、澱粉部分分解物１ｇあたり５０Ｕのグルコアミラーゼ（商品名『グルコチーム』、ナガセ生化学工業株式会社製）を加え、４０℃で２０時間インキュベートした。次いで、８０℃で１０分間インキュベートして酵素を失活させた後、実施例１−１におけると同様に、ろ紙でろ過して、さらに、合成吸着剤（商品名『ＨＰ１０』、三菱化学株式会社製）に負荷して吸着させ、水で洗浄して澱粉部分分解物を溶出させた後、２０乃至３０％（ｗ／ｗ）エタノール水溶液で溶出した画分を回収し、減圧乾固し、本発明の抗微生物剤に用いる没食子酸糖転移物を得た。
【００４４】
これを常法の高速液体クロマトグラフィー（ＨＰＬＣ）分析に供し、紫外部吸収を測定し組成比（モル比）を求めたところ、グルコース重合度が１の糖質部分を有する没食子酸糖転移物が８０％、グルコース重合度が２の糖質部分を有する没食子酸糖転移物が４％、グルコース重合度が３以上の糖質部分を有する没食子酸糖転移物が１％、未反応のまま残った没食子酸が１５％であった。
【００４５】
【実施例２】
＜没食子酸ｎ−プロピル糖転移物の調製＞
【００４６】
【実施例２−１】
実施例１−１の方法に準じて、１質量部の没食子酸ｎ−プロピル（関東化学株式会社製）を、１０質量部の澱粉部分分解物（ＤＥ８、商品名『パインデックス＃１』、松谷化学株式会社製）とともに２００質量部の水に添加して加熱溶解した。適量の水酸化ナトリウムを添加してｐＨ６．０に調整した後、澱粉部分分解物１ｇあたり２００Ｕのサイクロデキストリングルカノトランスフェラーゼ（株式会社林原生物化学研究所製）を加え、４０℃で２４時間インキュベートした。
その後、１００℃で２０分間インキュベートして酵素を失活させた後、ろ紙でろ過して、さらに、合成吸着剤（商品名『ＨＰ１０』、三菱化学株式会社製）に負荷して吸着させ、水で洗浄して澱粉部分分解物を溶出させた後、３５乃至５０％（ｗ／ｗ）エタノール水溶液で溶出した画分を回収し、減圧乾固し、本発明の抗微生物剤に用いる没食子酸ｎ−プロピル糖転移物を得た。
【００４７】
これを常法の高速液体クロマトグラフィー（ＨＰＬＣ）分析に供し、紫外部吸収を測定し組成比（モル比）を求めたところ、グルコース重合度が１の糖質部分を有する没食子酸ｎ−プロピル糖転移物が１４％、グルコース重合度が２の糖質部分を有する没食子酸ｎ−プロピル糖転移物が１０％、グルコース重合度が３以上の糖質を有する没食子酸ｎ−プロピル糖転移物が９％、未反応のまま残った没食子酸ｎ−プロピルが６７％であった。
【００４８】
【実施例２−２】
実施例１−２の方法に準じて、実施例２−１で得られたサイクロデキストリングルカノトランスフェラーゼ処理後の反応液に、適量の水酸化ナトリウムを添加してｐＨ４．５に調整した後、澱粉部分分解物１ｇあたり５０Ｕのグルコアミラーゼ（商品名『グルコチーム』、ナガセ生化学工業株式会社製）を加え、４０℃で２０時間インキュベートした。次いで、８０℃で１０分間インキュベートして酵素を失活させた後、実施例２−１におけると同様に、ろ紙でろ過して、さらに、合成吸着剤（商品名『ＨＰ１０』、三菱化学株式会社製）に負荷して吸着させ、水で洗浄して澱粉部分分解物を溶出させた後、３５乃至５０％（ｗ／ｗ）エタノール水溶液で溶出した画分を回収し、減圧乾固し、本発明の抗微生物剤に用いる没食子酸ｎ−プロピル糖転移物を得た。
【００４９】
これを常法の高速液体クロマトグラフィー（ＨＰＬＣ）分析に供し、紫外部吸収を測定し組成比（モル比）を求めたところ、グルコース重合度が１の糖質部分を有する没食子酸ｎ−プロピル糖転移物が７６％、グルコース重合度が２の糖質部分を有する没食子酸ｎ−プロピル糖転移物が７％、グルコース重合度が３以上の糖質部分を有する没食子酸ｎ−プロピル糖転移物が２％、未反応のまま残った没食子酸ｎ−プロピルが１５％であった。
【００５０】
【実施例３】
＜没食子酸オクチル糖転移物の調製＞
【００５１】
【実施例３−１】
実施例１−１の方法に準じて、１質量部の没食子オクチル（関東化学株式会社製）を、１０質量部の澱粉部分分解物（ＤＥ８、商品名『パインデックス＃１』、松谷化学株式会社製）とともに２００質量部の水に添加して加熱溶解した。適量の水酸化ナトリウムを添加してｐＨ６．０に調整した後、澱粉部分分解物１ｇあたり２００Ｕのサイクロデキストリングルカノトランスフェラーゼ（株式会社林原生物化学研究所製）を加え、４０℃で２４時間インキュベートした。その後、１００℃で２０分間インキュベートして酵素を失活させた後、ろ紙でろ過して、さらに、合成吸着剤（商品名『ＨＰ１０』、三菱化学株式会社製）に負荷して吸着させ、水で洗浄して澱粉部分分解物を溶出させた後、３５乃至５０％（ｗ／ｗ）エタノール水溶液で溶出した画分を回収し、減圧乾固し、本発明の抗微生物剤に用いる没食子酸オクチル糖転移物を得た。
【００５２】
これを常法の高速液体クロマトグラフィー（ＨＰＬＣ）分析に供し、紫外部吸収を測定し組成比（モル比）を求めたところ、グルコース重合度が１の糖質部分を有する没食子酸オクチル糖転移物が１４％、グルコース重合度が２の糖質部分を有する没食子酸オクチル糖転移物が５％、グルコース重合度が３以上の糖質を有する没食子酸オクチル糖転移物が７％、未反応のまま残った没食子酸オクチルが７４％であった。
【００５３】
【実施例３−２】
実施例１−２の方法に準じて、実施例３−１で得られたサイクロデキストリングルカノトランスフェラーゼ処理後の反応液に、適量の水酸化ナトリウムを添加してｐＨ４．５に調整した後、澱粉部分分解物１ｇあたり５０Ｕのグルコアミラーゼ（商品名『グルコチーム』、ナガセ生化学工業株式会社製）を加え、４０℃で２０時間インキュベートした。次いで、８０℃で１０分間インキュベートして酵素を失活させた後、実施例３−１におけると同様に、ろ紙でろ過して、さらに、合成吸着剤（商品名『ＨＰ１０』、三菱化学株式会社製）に負荷して吸着させ、水で洗浄して澱粉部分分解物を溶出させた後、３５乃至５０％（ｗ／ｗ）エタノール水溶液で溶出した画分を回収し、減圧乾固し、本発明の抗微生物剤に用いる没食子酸オクチル糖転移物を得た。
【００５４】
これを常法の高速液体クロマトグラフィー（ＨＰＬＣ）分析に供し、紫外部吸収を測定し組成比（モル比）を求めたところ、グルコース重合度が１の糖質部分を有する没食子酸オクチル糖転移物が７６％、グルコース重合度が２の糖質部分を有する没食子酸オクチル糖転移物が２％、グルコース重合度が３以上の糖質部分を有する没食子酸オクチル糖転移物が１％、未反応のまま残った没食子酸オクチルが２１％であった。
【００５５】
【実施例４】
＜カフェ酸糖転移物の調製＞
【００５６】
【実施例４−１】
実施例１−１の方法に準じて、１質量部のカフェ酸（和光純薬工業株式会社製）を、１０質量部の澱粉部分分解物（ＤＥ８、商品名『パインデックス＃１』、松谷化学株式会社製）とともに２００質量部の水に添加して加熱溶解した。適量の水酸化ナトリウムを添加してｐＨ６．０に調整した後、澱粉部分分解物１ｇあたり２００Ｕのサイクロデキストリングルカノトランスフェラーゼ（株式会社林原生物化学研究所製）を加え、４０℃で２４時間インキュベートした。その後、１００℃で２０分間インキュベートして酵素を失活させた後、ろ紙でろ過して、さらに、合成吸着剤（商品名『ＨＰ１０』、三菱化学株式会社製）に負荷して吸着させ、水で洗浄して澱粉部分分解物を溶出させた後、３５乃至５０％（ｗ／ｗ）エタノール水溶液で溶出した画分を回収し、減圧乾固し、本発明の抗微生物剤に用いるカフェ酸糖転移物を得た。
【００５７】
これを常法の高速液体クロマトグラフィー（ＨＰＬＣ）分析に供し、紫外部吸収を測定し組成比（モル比）を求めたところ、グルコース重合度が１の糖質部分を有するカフェ酸糖転移物が１９％、グルコース重合度が２の糖質部分を有するカフェ酸糖転移物が９％、グルコース重合度が３以上の糖質を有するカフェ酸糖転移物が７％、未反応のまま残ったカフェ酸が６５％であった。
【００５８】
【実施例４−２】
実施例１−２の方法に準じて、実施例４−１で得られたサイクロデキストリングルカノトランスフェラーゼ処理後の反応液に、適量の水酸化ナトリウムを添加してｐＨ４．５に調整した後、澱粉部分分解物１ｇあたり５０Ｕのグルコアミラーゼ（商品名『グルコチーム』、ナガセ生化学工業株式会社製）を加え、４０℃で２０時間インキュベートした。次いで、８０℃で１０分間インキュベートして酵素を失活させた後、実施例４−１におけると同様に、ろ紙でろ過して、さらに、合成吸着剤（商品名『ＨＰ１０』、三菱化学株式会社製）に負荷して吸着させ、水で洗浄して澱粉部分分解物を溶出させた後、２０乃至３０％（ｗ／ｗ）エタノール水溶液で溶出した画分を回収し、減圧乾固し、本発明の抗微生物剤に用いるカフェ酸糖転移物を得た。
【００５９】
これを常法の高速液体クロマトグラフィー（ＨＰＬＣ）分析に供し、紫外部吸収を測定し組成比（モル比）を求めたところ、グルコース重合度が１の糖質部分を有するカフェ酸糖転移物が７８％、グルコース重合度が２の糖質部分を有するカフェ酸糖転移物が３％、グルコース重合度が３以上の糖質部分を有するカフェ酸糖転移物が１％、未反応のまま残ったカフェ酸が１８％であった。
【００６０】
【実施例５】
＜ターミナリア ベリリカ抽出物の糖転移物の調製＞
モモタマナ属のターミナリア ベリリカ （Ｔｅｒｍｉｎａｌｉａ ｂｅｌｌｉｒｉｃａ）の果実部１質量部をブレンダーにより粉砕し、１０質量部の５０％（ｗ／ｗ）エタノール水溶液を加え、５０℃で６０分間攪拌した。ろ紙でろ過して、ろ液を採取し、遠心分離により固形成分を除去した上清を減圧乾燥して固形分を得た。この固形分１質量部を、実施例１−１の方法に準じて、１０質量部の澱粉部分分解物（ＤＥ８、商品名『パインデックス＃１』、松谷化学株式会社製）とともに２００質量部の水に添加して加熱溶解した。適量の水酸化ナトリウムを添加してｐＨ６．０に調整した後、澱粉部分分解物１ｇあたり２００Ｕのサイクロデキストリングルカノトランスフェラーゼ（株式会社林原生物化学研究所製）を加え、４０℃で２４時間インキュベートした。その後、１００℃で２０分間インキュベートして酵素を失活させた後、実施例１−２の方法に準じて、適量の水酸化ナトリウムを添加してｐＨ４．５に調整した後、澱粉部分分解物１ｇあたり５０Ｕのグルコアミラーゼ（商品名『グルコチーム』、ナガセ生化学工業株式会社製）を加え、４０℃で２０時間インキュベートした。次いで、８０℃で１０分間インキュベートして酵素を失活させた後、ろ紙でろ過して、ろ液を採取し、減圧乾固して、本発明の抗微生物剤に用いるターミナリア ベリリカ抽出物の糖転移物を得た。
【００６１】
このろ液を常法の高速液体クロマトグラフィー（ＨＰＬＣ）法に供し、没食子酸糖転移物の含量を測定したところ、固形分当り、有効成分である没食子酸糖転移物は８ｍｇ／ｇ、没食子酸は７ｍｇ／ｇであった。また、微量ながら、ペンタガロイルグルコースの糖転移物が検出された。
【００６２】
【実施例６】
＜藍抽出物の糖転移物＞
タデ藍（Ｐｏｌｙｇｏｎｕｍ Ｔｉｎｃｏｒｉｕｍ）の地上部１質量部をブレンダーで粉砕した後、水１．５質量部を加え、ミキサーで攪拌した。ろ紙でろ過して、ろ液を採取し、１２１℃で１０分間加熱し、遠心分離をして固形分を除去し、上清を回収した。次に、この上清を減圧乾燥して固形分を得た。これを実施例１−１の方法に準じて、藍抽出物１質量部に対し、１０質量部の澱粉部分分解物（ＤＥ８、商品名『パインデックス＃１』、松谷化学株式会社製）とともに２００質量部の水に添加して加熱溶解した。適量の水酸化ナトリウムを添加してｐＨ６．０に調整した後、澱粉部分分解物１ｇあたり２００Ｕのサイクロデキストリングルカノトランスフェラーゼ（株式会社林原生物化学研究所製）を加え、４０℃で２４時間インキュベートした。その後、１００℃で２０分間インキュベートして酵素を失活させた後、ろ紙でろ過して、ろ液を採取し、減圧乾固して、本発明の抗微生物剤に用いるタデ藍抽出物の糖転移物を得た。
【００６３】
このろ液を常法の高速液体クロマトグラフィー（ＨＰＬＣ）法に供し、カフェ酸糖転移物の含量を測定したところ、固形分当り、有効成分であるカフェ酸糖転移物は５ｍｇ／ｇ、カフェ酸が１２ｍｇ／ｇの濃度で含まれていた。
【００６４】
【実施例７】
＜口腔清浄剤＞
常法にしたがって、下記に示す口腔清浄剤の基本処方に対して、実施例１−１の方法により得た没食子酸糖転移物、実施例２−１の方法により得た没食子酸ｎ−プロピル糖転移物、実施例３−１の方法により得た没食子酸オクチル糖転移物、実施例４−１の方法により得たカフェ酸糖転移物、実施例５の方法により得たターミナリア ベリリカ抽出物の糖転移物、又は実施例６の方法により得たタデ藍抽出物の糖転移物をそれぞれ濃度１％（ｗ／ｗ）になるように配合して液状の口腔清浄剤を得た。
【００６５】
エタノール ４０質量部
グリセリン １５質量部
ポリオキシエチレン硬化ヒマシ油 １質量部
サッカリン 適量
香料 適量
クロルヘキシジン 適量
水 ４４重量部
【００６６】
本例の口腔洗浄剤は、通常のうがい薬と同様に常用することによって、口腔内のグラム陰性菌、グラム陽性菌、真菌、酵母などに抗微生物性を示す。例えば、バチラス属、ミコバクテリウム属、シュードモナス属、エッセリシア属、スタフィロコッカス属、ストレプトコッカス属、サルモネラ属、クレブシラ属、プロテウス属、ビブリオ属、シゲラ属、ヘリコバクター属、クラミジア属、アスペルギルス属及びカンディダ属の微生物の増殖を効果的に抑制し、それらが発症原因をなる各種疾患を効果的に治療・予防することができる。
【００６７】
【実施例８】
＜錠剤＞
常法にしたがって、マルトース粉末に、実施例１−２の方法により得た没食子酸糖転移物、実施例２−２の方法により得た没食子酸ｎ−プロピル糖転移物、実施例３−２の方法により得た没食子酸オクチル糖転移物、実施例４−２の方法により得たカフェ酸糖転移物、実施例５の方法により得たターミナリア ベリリカ抽出物の糖転移物、又は実施例６の方法により得たタデ藍抽出物の糖転移物をそれぞれ濃度１％（ｗ／ｗ）になるように配合した後、打錠機により、直径１ｃｍ、厚さ３．２ｍｍに打錠し、一錠０．５ｇの錠剤を得た。
【００６８】
本例の抗微生物剤は、通常の錠剤と同様に常用することによって、生体内で速やかに溶解し、生体における微生物の増殖を抑制し、かつ、生体における持続性が優れるので長時間抗微生物効果を発揮することから、グラム陰性菌、グラム陽性菌、真菌、酵母などに抗微生物性を示す。例えば、バチラス属、ミコバクテリウム属、シュードモナス属、エッセリシア属、スタフィロコッカス属、ストレプトコッカス属、サルモネラ属、クレブシラ属、プロテウス属、ビブリオ属、シゲラ属、ヘリコバクター属、クラミジア属、アスペルギルス属、カンディダ属の微生物が発症原因になる疾患を効果的に治療・予防することができる。
【００６９】
【実施例９】
＜練歯磨＞
常法にしたがって、下記に示す練歯磨の基本処方に対して、実施例１−１の方法により得た没食子酸糖転移物、実施例２−１の方法により得た没食子酸ｎ−プロピル糖転移物、実施例３−１の方法により得た没食子酸オクチル糖転移物、実施例４−１の方法により得たカフェ酸糖転移物、実施例５の方法により得たターミナリア ベリリカ抽出物の糖転移物、又は実施例６の方法により得たタデ藍抽出物の糖転移物を、それぞれ濃度１％（ｗ／ｗ）になるように配合した後、アルミニウムラミネートチューブへ１００ｇずつ充填してペースト状の抗微生物用の練歯磨を得た。
【００７０】
第二燐酸カルシウム ４２質量部
グリセリン １８質量部
カラギーナン ０．９質量部
ラウリル硫酸ナトリウム １．１質量部
香料 適量
サッカリン 適量
水 ３８質量部
【００７１】
本例の抗微生物用の練歯磨は、通常の練歯磨と同様に常用することによって、口腔におけるグラム陰性菌、グラム陽性菌、真菌、酵母などに抗微生物効果を示す。したがって、バチラス属、ミコバクテリウム属、シュードモナス属、エッセリシア属、スタフィロコッカス属、ストレプトコッカス属、サルモネラ属、クレブシラ属、プロテウス属、ビブリオ属、シゲラ属、ヘリコバクター属、クラミジア属、アスペルギルス属、カンディダ属の微生物が発症原因になる疾患を効果的に治療・予防することができる。
【００７２】
【実施例１０】
＜トローチ剤＞
常法にしたがって、下記に示すトローチ剤の基本処方に対して、実施例１−１の方法により得た没食子酸糖転移物、実施例２−１の方法により得た没食子酸ｎ−プロピル糖転移物、実施例３−１の方法により得た没食子酸オクチル糖転移物、実施例４−１の方法により得たカフェ酸糖転移物、実施例５の方法により得たターミナリア ベリリカ抽出物の糖転移物、又は実施例６の方法により得たタデ藍抽出物の糖転移物をそれぞれ濃度１％（ｗ／ｗ）になるように配合し、成形して固状の抗微生物用のトローチ剤を得た。
【００７３】
マルトース ３５質量部
澱粉 ３４質量部
結晶セルロース １０質量部
ヒドロキシプロピルメチルセルロース １０質量部
ステアリン酸マグネシウム １質量部
【００７４】
本例の抗微生物用のトローチ剤は、通常のトローチ剤と同様に常用することによって、口腔内で速やかに溶解し、口腔内におけるグラム陰性菌、グラム陽性菌、真菌、酵母などに抗微生物効果を示す。したがって、バチラス属、ミコバクテリウム属、シュードモナス属、エッセリシア属、スタフィロコッカス属、ストレプトコッカス属、サルモネラ属、クレブシラ属、プロテウス属、ビブリオ属、シゲラ属、ヘリコバクター属、クラミジア属、アスペルギルス属、カンディダ属の微生物が発症原因になる疾患を効果的に治療・予防することができる。
【００７５】
【実施例１１】
＜液剤＞
常法にしたがって、イソロイシンを４質量％含有する注射用精製水に、実施例１−２の方法により得た没食子酸糖転移物、実施例２−２の方法により得た没食子酸ｎ−プロピル糖転移物、実施例３−２の方法により得た没食子酸オクチル糖転移物、実施例４−２の方法により得たカフェ酸糖転移物、実施例５の方法により得たターミナリア ベリリカ抽出物の糖転移物、又は実施例６の方法により得たタデ藍抽出物の糖転移物を濃度１％（ｗ／ｗ）になるように溶解させた後、精密濾過、容器に充填して液状の抗微生物効果を有する液剤を得た。
【００７６】
本例の抗微生物効果を有する液剤は、注射用剤、経粘皮用剤、経鼻腔用剤、経口用剤などとして、気道、鼻腔、口腔、視覚器、消化器、泌尿器、生殖器などにおけるグラム陰性菌、グラム陽性菌、真菌、酵母などに抗微生物効果を示す。したがって、バチラス属、ミコバクテリウム属、シュードモナス属、エッセリシア属、スタフィロコッカス属、ストレプトコッカス属、サルモネラ属、クレブシラ属、プロテウス属、ビブリオ属、シゲラ属、ヘリコバクター属、クラミジア属、アスペルギルス属、カンディダ属の微生物が発症原因になる疾患を効果的に治療・予防することができる。
【００７７】
【実施例１２】
＜軟膏＞
常法にしたがって、実施例１−１の方法により得た没食子酸糖転移物、実施例２−１の方法により得た没食子酸ｎ−プロピル糖転移物、実施例３−１の方法により得た没食子酸オクチル糖転移物、実施例４−１の方法により得たカフェ酸糖転移物、実施例５の方法により得たターミナリア ベリリカ抽出物の糖転移物、又は実施例６の方法により得たタデ藍抽出物の糖転移物を濃度１％（ｗ／ｗ）になるように適量の燐酸緩衝生理食塩水（ｐＨ７．２）に溶解し、その水溶液１００質量部を無水結晶マルトース４９質量部と均一に配合した後、さらに４５０質量部の白色ワセリンと練り合わせて抗微生物用の軟膏を得た。
【００７８】
本例の抗微生物用の軟膏は、皮膚外用剤、経粘皮用剤などとして、皮膚、泌尿器、生殖器などにおけるグラム陰性菌、グラム陽性菌、真菌、酵母などに抗微生物効果を示す。したがって、バチラス属、ミコバクテリウム属、シュードモナス属、エッセリシア属、スタフィロコッカス属、ストレプトコッカス属、サルモネラ属、クレブシラ属、プロテウス属、ビブリオ属、シゲラ属、ヘリコバクター属、クラミジア属、アスペルギルス属、カンディダ属の微生物が発症原因になる疾患を効果的に治療・予防することができる。
【００７９】
【実施例１３】
＜点眼剤＞
常法にしたがって、下記に示す点眼剤の基本処方に対して、実施例１−２の方法により得た没食子酸糖転移物、実施例２−２の方法により得た没食子酸ｎ−プロピル糖転移物、実施例３−２の方法により得た没食子酸オクチル糖転移物、実施例４−２の方法により得たカフェ酸糖転移物、実施例５の方法により得たターミナリア ベリリカ抽出物の糖転移物、又は実施例６の方法により得たタデ藍抽出物の糖転移物を濃度１％（ｗ／ｗ）になるように配合して液状の抗微生物用の点眼剤を得た。
【００８０】
トレハロース（２含水物） ２０質量部
塩化ナトリウム ０．０４質量部
塩化カリウム ０．０２質量部
燐酸２水素ナトリウム ０．０３質量部
硼砂 ０．０４質量部
塩化ベンザルコニウム ０．００４質量部
精製水 ８０質量部
【００８１】
本例の抗微生物用の点眼剤は、通常の点眼剤と同様に常用することによって、視覚器におけるグラム陰性菌、グラム陽性菌、真菌、酵母などに抗微生物効果を示す。したがって、バチラス属、ミコバクテリウム属、シュードモナス属、エッセリシア属、スタフィロコッカス属、ストレプトコッカス属、サルモネラ属、クレブシラ属、プロテウス属、ビブリオ属、シゲラ属、ヘリコバクター属、クラミジア属、アスペルギルス属、カンディダ属の微生物が発症原因になる疾患を効果的に治療・予防することができる。
【００８２】
【発明の効果】
以上説明したとおり、本発明の抗微生物剤は、生体に対する毒性が低いうえ、水溶性に優れ生体内における持続性に優れるので、注射又は経皮、経粘皮、経鼻腔若しくは経口経路など生体に適用することによって、生体にとって好ましくない微生物の増殖を著明に抑制することができる。したがって、鼻腔、口腔、食道や気道、さらには視覚器、消化器、泌尿器、生殖器などへ侵入した微生物が発症原因となる、例えば、肺炎などの呼吸器系疾患や、動脈硬化、虚血性心疾患などの循環器系疾患、結膜炎などの視覚器系疾患、直腸炎などの消化器系疾患、尿道炎などの泌尿器系疾患、副睾丸炎、卵管炎、子宮頚管炎、切迫早産、卵管の炎症、癒着、閉塞などの生殖器系疾患、食中毒などの治療、予防に著効を発揮する。
とりわけ、本発明の抗微生物用口腔用剤は、抗生物質を有効成分とする経口用剤とは違って、耐性菌の出現を招来したり、長期間連用しても大腸における有用な菌叢を破壊する恐れが少ないないので、食欲不振、嘔吐、下痢、便秘、膨満感を生じたり、倦怠感、発湿などの副作用を惹起する恐れが少ない。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to an antimicrobial agent, and more specifically, one or more selected from a glycosyl transfer product of hydroxybenzoic acid, a glycosyl transfer product of hydroxycinnamic acid, and derivatives thereof, and salts thereof. The present invention relates to an antimicrobial agent contained as an active ingredient.
[0002]
[Prior art]
In recent years, a clean living environment has been demanded, and antimicrobial agents are used in many articles such as foods, clothes, cosmetics, daily necessities, and building materials in addition to pharmaceutical applications. However, antimicrobial agents contain metal compounds and organic compounds as active ingredients, and many of them have a risk of being harmful to the environment and the human body. For example, halogenated phenols such as hexachlorophene and bithionol, halogenated salicylanilide, boric acid, hydrogen peroxide, mercury compounds, formaldehyde, etc. are now highly sensitive to skin and respiratory organs. Use is prohibited. In particular, formaldehyde is contained in a large amount in building materials and is well known as a causative substance of sick house syndrome. In addition, antibiotics used as pharmaceuticals have a risk of destroying useful flora in the large intestine and generating resistant bacteria.
[0003]
Currently, antimicrobial agents with high safety are required, and plant extracts or components contained therein are being used as antimicrobial agents for foods and food additives from the viewpoint of high safety. For example, it has been proposed to use hinokitiol, Shoso bamboo extract, green tea extract, indigo extract, propolis extract and the like as antimicrobial agents.
However, although these extracts are excellent in safety and antimicrobial effect, they have a problem that handling property is not good because the in vivo sustainability and water solubility of the compound contained as an active ingredient are poor. .
[0004]
[Problems to be solved by the invention]
In view of such a situation, an object of the present invention is to provide an antimicrobial agent that has a safe and sufficient antimicrobial effect on the living body and has good handleability.
[0005]
[Means for Solving the Problems]
In order to solve the above-mentioned problems, the present inventors have conducted research focusing on hydroxyphenol compounds contained in plant extracts. As a result, gallic acid, which is a kind of hydroxybenzoic acid, and hydroxysilicate Caffeic acid, a kind of cinnamate, has an advantageous characteristic that it is relatively superior in antimicrobial effect among plant-derived antimicrobial compounds and has low toxicity to the human body. It has been found that there is a problem that sustainability in vivo is extremely low. Therefore, when glycated products of gallic acid or caffeic acid were prepared and the antimicrobial effect and persistence in vivo were examined, these glycosylated products were compared with gallic acid or caffeic acid that had not been transglycosylated. Although the antimicrobial effect is somewhat inferior, the present inventors have found that the compound has a level of activity that can be used as an antimicrobial agent, is excellent in water solubility, and has good handleability, thereby completing the present invention.
[0006]
That is, the present invention contains, as an active ingredient, one or more selected from a glycosylated product of hydroxybenzoic acid, a glycosylated product of hydroxycinnamic acid, and derivatives thereof, and salts thereof. The problem is solved by providing an antimicrobial agent.
[0007]
DETAILED DESCRIPTION OF THE INVENTION
Hydroxybenzoic acid as referred to in the present invention is a component constituting the plant component tannin, and its sugar transfer product is used as an active ingredient of the antimicrobial agent of the present invention. In this specification, a kind of hydroxybenzoic acid gallic acid (3,4,5-trihydroxybenzoic acid, molecular weight of about 170) will be described. However, as long as the effect of the present invention is exhibited, other hydroxybenzoic acids are used. can do. As a method for preparing gallic acid used in the present invention, it is obtained by extracting from a plant rich in gallic acid or a derivative thereof, and hydrolyzing with an acid, alkali, heat, enzyme, etc., if necessary. Can do. The gallic acid used in the present invention can be obtained by a chemical synthesis method in addition to a method of extracting and purifying from a plant body regardless of its origin and preparation method, and it is optional to use a commercially available product. . In addition, a salt form can be used as long as the effects of the present invention are not impaired.
[0008]
Hydroxycinnamic acid as referred to in the present invention is a kind of plant component, and its glycosylated product is used as an active ingredient of the antimicrobial agent of the present invention. In this specification, a kind of hydroxycinnamic acid, caffeic acid (3,4-dihydroxycinnamic acid, molecular weight of about 180) will be described. However, as long as the effect of the present invention is exhibited, other hydroxycinnamic acids are used. can do. As a method for preparing caffeic acid used in the present invention, it is obtained by extracting from a plant rich in caffeic acid or a derivative thereof, and hydrolyzing with an acid, alkali, heat, enzyme or the like, if necessary. Can do. Caffeic acid used in the present invention is not limited to the method of extraction and purification from plant bodies regardless of its origin and preparation method, but also chemical synthesis, for example, 3,4-dihydroxybenzaldehyde, acetic anhydride and potassium acetate It can also be obtained by a method of synthesizing by reacting, and it is optional to use a commercially available product. In addition, a salt form can be used as long as the effects of the present invention are not impaired.
[0009]
Glycosyl transfer products and derivatives thereof of gallic acid used for the antimicrobial agent of the present invention, and glycosyl transfer products of caffeic acid and derivatives thereof are gallic acid or derivatives thereof by chemical or enzymatic glycosyl transfer reaction, and A sugar is transferred to caffeic acid or a derivative thereof via a hydroxyl group thereof. In the case where the sugar having undergone transglycosylation is glucose, examples of the typical gallic acid glycosyl transfer product include those of chemical formulas 1 to 3, and examples of the caffeic acid glycosyl transfer product include those of chemical formulas 4 and 5. In the formula, m and n represent an integer of 0 or more, and R represents a hydrogen atom or a hydrocarbon group. In Chemical Formulas 1 to 5, the bond between glucose is 1,4 bond, but it may be 1,3 bond or 1,6 bond. Further, in Chemical Formulas 1 to 5, although α-glucoside bonds are exemplified as the binding mode between the saccharide and gallic acid or caffeic acid, it is also possible to select β-glucoside bonds as necessary. . In addition, the caffeic acid sugar transfer product or the gallic acid sugar transfer product can be in the form of a salt as long as the effects of the present invention are not impaired.
[0010]
[Chemical 1]
Chemical formula 1:

(In the formula, n represents an integer of 0 or more, and R represents a hydrogen atom or a hydrocarbon group)
[0011]
[Chemical 2]
Chemical formula 2:

(In the formula, n represents an integer of 0 or more, and R represents a hydrogen atom or a hydrocarbon group)
[0012]
[Chemical Formula 3]
Chemical formula 3:

(In the formula, m and n represent an integer of 0 or more, and R represents a hydrogen atom or a hydrocarbon group)
[0013]
[Formula 4]
Chemical formula 4:

(In the formula, n represents an integer of 0 or more, and R represents a hydrogen atom or a hydrocarbon group)
[0014]
[Chemical formula 5]
Chemical formula 5:

(In the formula, n represents an integer of 0 or more, and R represents a hydrogen atom or a hydrocarbon group)
[0015]
As the antimicrobial agent of the present invention, a gallic acid sugar transfer product and a caffeic acid sugar transfer product derivative can be used. The order of sugar transfer and derivatization to gallic acid and caffeic acid does not matter. As derivatives of gallic acid transfer products and caffeic acid transfer products, compounds that can be hydrolyzed relatively easily by treatment with acid, alkali, heating, etc. to produce gallic acid or caffeic acid are used. They can be natural or artificially synthesized. For example, as derivatives of gallic acid that exist in nature, a compound in which 1 to 5 molecules of gallic acid are ester-linked to one molecule of glucose, which is generically referred to as hydrolyzable tannin, such as galloylglucose, kebric acid, and ketebraic acid On the other hand, chlorogenic acid and the like can be exemplified as a derivative of caffeic acid. Since the glycosylated products of these derivatives produce a glycosylated product of gallic acid or a glycosylated product of caffeic acid after hydrolysis and exhibit an antimicrobial effect, they can be used as a component of the antimicrobial agent of the present invention. . When transglycosylating gallic acid or a derivative of caffeic acid, a compound having a hydroxyl group capable of transglycosylation to gallic acid and caffeic acid moiety is preferred, and sugars other than gallic acid or caffeic acid moiety are also preferred. A compound having no transferable hydroxyl group is preferred. As such a derivative, for example, pentagalloylglucose (1,2,3,4,6-pentakis-O-galloyl-β-D-glucose) which is a kind of galloylglucose represented by Chemical Formula 6 can be mentioned. .
[0016]
[Chemical 6]
Chemical formula 6:

[0017]
Furthermore, as derivatives of gallic acid and caffeic acid, derivatives having appropriate modifications can be used in order to improve the antimicrobial effect of gallic acid and caffeic acid. Examples of such derivatives include compounds in which a hydrocarbon group is ester-bonded to a carboxyl group of gallic acid and caffeic acid. The hydrocarbon group to be ester-bonded may be a chain hydrocarbon group or a cyclic hydrocarbon group, and may be a saturated hydrocarbon group or an unsaturated hydrocarbon group. Preferred hydrocarbon groups in the present invention are chain hydrocarbon groups, so-called aliphatic hydrocarbon groups. An ester derivative of gallic acid or caffeic acid and an aliphatic hydrocarbon group (hereinafter sometimes simply referred to as an ester derivative) has a feature that antimicrobial properties are improved according to the number of carbon atoms of the aliphatic hydrocarbon group. And is advantageously used as the antimicrobial agent of the present invention. In addition, although ester derivatives of gallic acid or caffeic acid and an aliphatic hydrocarbon group tend to become less water-soluble as the number of carbon atoms in the hydrocarbon group increases, water solubility is improved by sugar transfer. The number of carbon atoms of the hydrocarbon group may be appropriately selected according to the method of using the antimicrobial agent of the present invention, the form thereof, the target microorganism, and the like, and is usually in the range of 1 to 12 carbon atoms, preferably 3 to 8 carbon atoms. Those obtained by sugar transfer to ester derivatives with a range of hydrocarbon groups are used as the antimicrobial agent of the present invention. Since ester derivatives with hydrocarbon groups having more than 12 carbon atoms have significantly reduced water solubility, the water solubility is not sufficiently improved even by sugar transfer, and the antimicrobial agent of the present invention is applied to a living body. May not be preferable. In addition, n-propyl gallate, which is an ester derivative of gallic acid and an aliphatic hydrocarbon group having 3 carbon atoms, has high antimicrobial properties despite the small number of carbon atoms in the hydrocarbon group. It is particularly advantageously used as the antimicrobial agent of the invention.
[0018]
As a method for obtaining a glycosylated product of gallic acid, a glycosylated product of caffeic acid, and derivatives thereof (hereinafter sometimes simply referred to as “glycosylated product of gallic acid” or “glycosylated product of caffeic acid”). Is not particularly limited, and can be produced by a chemical synthesis method or an enzymatic reaction method. Among these, since the enzyme reaction method using glycosyltransferase is simple, it can be advantageously used in the present invention. For example, dextrin, cyclodextrin, starch or a mixture thereof is added to gallic acid or a derivative thereof, or caffeic acid or a derivative thereof, and this is added to, for example, a glycosyltransferase produced by a microorganism belonging to the genus Bacillus, for example, Bacillus・ Cyclodextrin glucanotransferase derived from stearothermophilus (EC 2.4.1.19) is allowed to act on sugar, and then, if desired, glucoamylase and / or β-amylase is allowed to act on the enzyme reaction solution. The method of letting it be mentioned. According to this method, it is possible to adjust the degree of polymerization of the saccharide portion of the glycated gallic acid transfer product and the caffeic acid saccharide transfer product.
[0019]
Moreover, as a method for obtaining a glycosylated product of gallic acid or a glycosylated product of caffeic acid, an extract of a plant body containing these can be glycosylated as it is or at an appropriate stage of the purification process. In this case, gallic acid or a derivative thereof, or caffeic acid or a derivative thereof, and some of the other components become a sugar transfer product, which also improves the water solubility of those components, so the water solubility of the extract It can be expected to greatly improve As a plant containing gallic acid or a derivative thereof, or caffeic acid or a derivative thereof, green tea, black tea, coffee, and the like are common. In addition, plants belonging to the genus Momotamana, Tade indigo (Polygonum genus) and the like can be mentioned. However, it is not limited to these.
[0020]
The antimicrobial agent of the present invention includes gallic acid or a derivative thereof, or a caffeic acid or a derivative thereof (hereinafter referred to as a non-glycosyl transfer product derived from the production method), in addition to a glycated sugar-transferred product or a caffeic acid-transferred product May be simply referred to as “unglycosylated product”). The unglycosylated product can be removed from the glycosylated product by a technique such as high performance liquid chromatography (HPLC), but it can also be added to the antimicrobial agent of the present invention while containing the unglycosylated product. If necessary, a non-glycosylated product corresponding to the glycosylated product can be blended with the antimicrobial agent of the present invention. In this case, the content of the glycosylated product is usually 10% or more, preferably 20% or more, and more preferably 50% or more, in molar ratio with respect to the sum of the glycosylated product and the unglycosylated product. If the ratio of the glycosylated product is less than 10%, the effect of improving the in vivo sustainability and water solubility cannot be sufficiently expected, which may be undesirable.
[0021]
Further, in the glycated sugar transfer product or caffeic acid transfer product used in the present invention, in addition to glucose, monosaccharides such as galactose, mannose, and fucose, amino sugars such as glucosamine and galactosamine, and the like can be transferred to sugar. You can choose. The degree of polymerization of the saccharide moiety in the glycosylated product is preferably selected from 1 to 5, more preferably 1 to 3, and most preferably 1. The higher the degree of polymerization of the saccharide part, the better the effect of improving the in vivo persistence and water solubility of gallic acid and caffeic acid. Tend to decrease. Therefore, when an immediate effect is expected, a sugar transfer product having a saccharide portion having a degree of polymerization exceeding 5 may not be preferable.
[0022]
The antimicrobial agent of the present invention contains a gallic acid transfer product and / or a caffeic acid transfer product as an active ingredient, and optionally gallic acid or a derivative thereof, and / or caffeic acid or a derivative thereof. Can be blended. In general, one or more of the components for constituting an injectable agent, an external skin agent, a transmucosal agent, a nasal agent, or an oral agent can be blended.
[0023]
The antimicrobial agent of the present invention is prepared in the form of, for example, liquid, paste or solid injection, external skin preparation, transmucosal agent, nasal cavity agent, oral agent, etc., depending on the use. Specifically, for oral preparations such as injections, eye drops, suppositories, ointments, oral cleansing agents such as toothpastes and mouthwashes, and oral bactericides in the dental field. Form is preferred. Therefore, the antimicrobial agent of the present invention includes powders, fine granules, pills, tablets, capsules, troches, limonades, elixirs, syrups, fragrances, suspensions, emulsions, dipping agents, decoctions, Commonly used in tinctures, extracts, flow extracts, spirits, liniments, lotions, ointments, poultices, plasters, suppositories, eye drops, eye ointments, injections, for example, abrasives, Foaming agent, wetting agent, binder, flavoring agent, sweetener, preservative, fluorine compound, enzyme agent, deodorant, and further, excipient, ointment base, solubilizer, flavoring agent, flavoring agent, coloring Use in combination with one or more preparations such as an agent, an emulsifier, a propellant, etc. In addition, it is widely used in drugs of such dosage forms without departing from the object of the present invention, for example, hemostatic agents, anti-inflammatory agents, tissue activators, antimicrobial agents, antibiotics, interferon α, interferon Combination with one or a plurality of other medicinal components including β and interferon γ is not hindered. In addition, as a preparation for injections, transmucosal agents, nasal cavity, oral preparations, it has the property of stabilizing glycated acid transfer products and caffeic acid transfer products, for example, Sugar, sugar alcohols such as sucrose, glucose, maltose, trehalose, fructose, cyclic tetrasaccharide, mannitol, maltitol, as well as alanine, arginine, asparagine, aspartic acid, citrulline, cysteine, cystine, glutamine, glycine, histidine, isoleucine , Leucine, lysine, methionine, ornithine, phenylalanine, phenylglycine, proline, serine, threonine, tryptophan, tyrosine, valine, hydroxyproline, γ-carboxyglutamate, O-phosphoserine, β-alanine, α-aminobutyric acid, γ- Amino dairy , Α-aminoisobutyric acid, 4- (4-aminophenyl) butyric acid, aminophenylbutyric acid, aminobenzoic acid, 4-aminohippuric acid, aminomethylbenzoic acid, ε-aminocaproic acid, 7-aminoheptanoic acid, β-asparagine Preferred are amino acids such as acid, γ-glutamic acid, ε-lysine, methionine sulfone, norleucine, norvaline, ornithine, d-ornithine, p-nitro-phenylalanine, hydroxyproline, thioproline, or derivatives or oligopeptides of these amino acids, if necessary Accordingly, these are used in combination.
[0024]
The antimicrobial spectrum of the antimicrobial agent of the present invention is relatively wide and exhibits an antimicrobial effect against many microorganisms such as gram-negative bacteria, gram-positive bacteria, fungi and yeast. Examples of the microorganisms targeted by the antimicrobial agent of the present invention include, for example, Bacillus, Mycobacterium, Pseudomonas, Esseria, Staphylococcus, Streptococcus, Salmonella, Klebsiella, Proteus, Vibrio, Examples include microorganisms belonging to the genus Shigella, Helicobacter, Chlamydia, Aspergillus, Candida and the like.
[0025]
The amount of the gallic acid glycosylated product and / or the caffeic acid glycosylated product to be blended with the antimicrobial agent of the present invention may be appropriately determined in consideration of its dosage form, method of use, etc. Alternatively, in terms of caffeic acid, the total amount of gallic acid and caffeic acid is 0.001 to 100% (w / w), preferably 0.01 to 100% (w / w), more preferably 0.1 to It is selected from the range of 100% (w / w). The dose depends on the application target and administration route of the antimicrobial agent, but the total amount of gallic acid and caffeic acid is usually 1 ng to 1 g / day / adult in terms of gallic acid or caffeic acid, Preferably, 10 ng to 100 mg / day / adult.
[0026]
The antimicrobial agent of the present invention can be applied to various kinds of living organisms by injection or transdermal, transmucosal, nasal agent or oral route at such a dose at a frequency of once or more per day or week. Because it significantly suppresses the growth of pathogenic microorganisms, pathogenic microorganisms that have entered the nasal cavity, oral cavity, respiratory tract, visual organs, digestive organs, urinary organs, genitals, etc. cause onset, for example, respiration such as pneumonia Systemic diseases, circulatory system diseases such as arteriosclerosis, ischemic heart disease, visual system diseases such as conjunctivitis, digestive system diseases such as proctitis, urinary system diseases such as urethritis, accessory testicularitis, fallopian tube It is effective for the treatment and prevention of genital system diseases such as inflammation, cervicitis, imminent premature labor, fallopian tube inflammation, adhesion, obstruction, and food poisoning. In particular, the antimicrobial agent of the present invention, unlike oral preparations containing antibiotics as an active ingredient, may cause the appearance of resistant bacteria or destroy the useful flora in the large intestine even after prolonged use. It does not cause loss of appetite, vomiting, diarrhea, constipation, bloating, or causing side effects such as malaise and moisture.
[0027]
As described above, the glycosylated product of gallic acid and / or the glycosylated product of caffeic acid have the advantage of being excellent in sustainability and water solubility in a living body. These characteristics are also exhibited when a compound having poor water solubility, such as a gallic acid ester derivative having an aliphatic hydrocarbon group having 3 or more carbon atoms, caffeic acid and a caffeic acid ester derivative, is transglycosylated. In addition, it is known that three adjacent hydroxyl groups of gallic acid form a complex with iron ions and discolor, and therefore, an effect of preventing discoloration of gallic acid can be expected by sugar transfer.
[0028]
Hereinafter, the properties and effects of the antimicrobial agent according to the present invention will be described based on experimental examples.
[0029]
[Experiment 1]
<Preparation of sugar transfer product of gallic acid, gallic acid ester derivative and caffeic acid>
1 part by weight of gallic acid, methyl gallate, ethyl gallate, n-propyl gallate, octyl gallate, dodecyl gallate (manufactured by Kanto Chemical Co., Ltd.) or caffeic acid (manufactured by Wako Pure Chemical Industries, Ltd.) Each of them was added to 200 parts by mass of water together with 10 parts by mass of a partially decomposed starch (DE8, trade name “Paindex # 1”, manufactured by Matsutani Chemical Co., Ltd.) and dissolved by heating. After adjusting the pH to 6.0 by adding an appropriate amount of sodium hydroxide, 200 U cyclodextrin glucanotransferase (produced by Hayashibara Biochemical Laboratories) was added per 1 g of the partially decomposed starch and incubated at 40 ° C. for 24 hours. . Then, after incubating at 100 ° C. for 20 minutes to inactivate the enzyme, an appropriate amount of sodium hydroxide was added to adjust the pH to 4.5, and then 50 U of glucoamylase (trade name “gluco” Team ”, manufactured by Nagase Seikagaku Corporation), and incubated at 40 ° C. for 20 hours. Next, the enzyme was inactivated by incubating at 80 ° C. for 10 minutes, followed by filtration with filter paper. In each reaction solution, 200 ml of synthetic adsorbent (trade name “HP10”, Mitsubishi Chemical) per gram of gallic acid, methyl gallate, ethyl gallate, n-propyl gallate, octyl gallate, dodecyl gallate, or caffeic acid Loaded onto a column packed with Co., Ltd., washed with water, further washed with 5% (w / w) aqueous ethanol solution, and 15 to 50% (w / w) depending on the hydrophobicity of each sample. w) Elution with an aqueous ethanol solution. After these were dried under reduced pressure, redissolved in water, the same operation was repeated with the same column, the eluate was dried under reduced pressure, and the sugar transfer product of gallic acid used for the antimicrobial agent of the present invention, gallic acid A methyl glycosyl transfer product, an ethyl gallate glycosyl transfer product, an n-propyl gallate glycosyl transfer product, an octyl gallate glycosyl transfer product, a dodecyl gallate glycosyl transfer product, and a caffeic acid glycosyl transfer product were obtained. When these were measured for ultraviolet absorption by conventional high performance liquid chromatography (HPLC) analysis and the composition ratio (molar ratio) was determined, all the glycosylated products prepared were monoglucosides, and their purity was All were over 95%.
1U of cyclodextrin glucanotransferase is prepared by adding 0.2 ml of enzyme solution to 5 ml of 0.3% (w / w) soluble starch powder solution dissolved in 20 mM acetate buffer (pH 5.5) containing 1 mM calcium chloride. The mixture was reacted at 40 ° C. for 10 minutes, 0.5 ml of this reaction solution was added to 15 ml of 0.02N sulfuric acid to stop the reaction, 0.2 ml of 0.1N iodine solution was added thereto, and the absorbance at 660 nm was then added. Is defined as the amount of enzyme that reduces the absorbance by 10%. In addition, 1U of glucoamylase is added to 0.2 ml of enzyme solution to 5 ml of 1.0% (w / w) soluble starch powder solution (20 mM acetate buffer, pH 4.5), and reacted at 40 ° C. for 10 minutes. When the amount of produced reducing sugar is measured by the Somogy-Nelson method, it is defined as the amount of enzyme that liberates reducing power corresponding to 1 μmol of glucose per minute at 40 ° C.
[0030]
[Experimental example 2]
<Water-solubility of sugar transfer products of gallic acid, gallic acid ester derivatives and caffeic acid>
The solubility of the glycosylated product obtained in Experimental Example 1 in water at 37 ° C. was measured and compared with each of the unglycosylated products. The results are shown in Table 1.
[0031]
[Table 1]

[0032]
From the results shown in Table 1, gallic acid, gallic acid ester derivatives, and caffeic acid sugar transfer products are superior in water solubility to their respective non-sugar transfer products, and in particular, gallic acid n-, which is relatively poor in water solubility. For propyl, octyl gallate, dodecyl gallate and caffeic acid, the effect of improving water solubility was significant. Separately, a similar experiment was performed on a glycosylated product in which the degree of polymerization of the transferred sugar exceeded 1, and the higher the degree of polymerization of the sugar-transferred saccharide part, the better the water-solubility effect. It was.
[0033]
[Experiment 3]
<Antimicrobial effect of various sugar transfer products>
Antimicrobial effects of various glycosylated products prepared in Experimental Example 1, a glycosylated product of Terminaria berylica extract prepared in Example 5 described later, and a glycosylated product of Tade indigo extract prepared in Example 6 described later The minimum growth inhibitory concentration (MIC) of the microorganisms listed in Table 2 was measured according to a conventional method. Moreover, each non-sugar transfer thing was used as a control. The results are shown in Table 2. In addition, the gram number of methyl gallate, ethyl gallate, n-propyl gallate, octyl gallate, dodecyl gallate and its sugar transfer product in Table 2 indicates the value in terms of gallic acid, and the terminaria berylica extract and Tade indigo extracts and their sugar transfer products are shown in grams of solids.
[0034]
[Table 2]

[0035]
From the results in Table 2, it was found that the minimum growth inhibitory concentration for microorganisms was reduced in the range of equivalent to 1/32 as a result of sugar transfer, but the antimicrobial effect was retained. On the other hand, in the case of Terminaria berylica extract and indigo extract, the antimicrobial effect is unexpectedly considered considering that the content of the glycated product of gallic acid or caffeic acid as an active ingredient is about 1% per solid content of the extract. It was excellent and it was assumed that the effects of other components were added. In addition, when the same test was conducted with a high degree of polymerization of the saccharide portion of the sugar transfer product, a tendency was observed that the antimicrobial activity decreased as the degree of polymerization increased.
[0036]
[Experimental Example 4]
<Durability in vivo>
Various glycosylated products prepared in Experimental Example 1 were prepared in 0.1 mg / ml aqueous solution in terms of gallic acid or caffeic acid, and sterilized by filtration. On the other hand, the glycosyl transfer product of Terminaria berylica extract prepared in Example 5 described later and the glycosyl transfer product of Tade indigo extract prepared in Example 6 described later have 0.1 mg of gallic acid or caffeic acid contained therein. The solution was prepared to be a / ml aqueous solution, and sterilized by filtration. These were each injected 1 ml (0.1 mg) into the femoral vein of a 5-week-old female Wistar rat. After standing for a certain period of time, blood was collected over time from the tail vein of the rat, and the contents of gallic acid and caffeic acid in the serum were measured by a conventional liquid high-performance chromatography method. Moreover, it tested similarly using each non-sugar transfer thing as a control | contrast. As a result, unglycosylated products can no longer be detected in the blood approximately 2 hours after intravenous injection, whereas in the case of glycosylated products, gallic acid, gallate derivatives or cafes from blood until 8 hours after intravenous injection. The acid could be detected. This result shows that the glycosylated product is superior in sustainability in vivo to the unsuccessed product. At the same time, when the same test was performed on a sugar transfer product having a high degree of polymerization of the saccharide moiety, the sustainability of gallic acid, gallic acid ester derivatives and caffeic acid in vivo increased as the degree of polymerization increased. A trend was observed.
[0037]
[Experimental Example 5]
<Acute toxicity test>
The various glycosylated products prepared in Experimental Example 1 were administered percutaneously, orally or intraperitoneally to 10 male mice each 8 weeks old according to a conventional method. As a result, these LDs₅₀Was about 100 mg / kg (mouse body weight) or more for any route of administration. This result shows that the glycosylated product of gallic acid and the glycosylated product of caffeic acid are safe as pharmaceuticals premised on administration to humans.
[0038]
From the above results, the glycated and / or caffeic acid glycosylated product of gallic acid is inferior to the antimicrobial effect of these unglycosylated products, but has an antimicrobial effect exceeding the level useful as an antimicrobial agent. Furthermore, it was excellent in in vivo sustainability and water solubility. The characteristics possessed by these sugar transfer products indicate that the antimicrobial agent of the present invention is advantageous as an antimicrobial agent applied to a living body.
[0039]
Hereinafter, embodiments of the present invention will be described based on examples.
[0040]
[Example 1]
<Preparation of gallic acid sugar transfer product>
[0041]
Example 1-1
1 part by weight of gallic acid (manufactured by Kanto Chemical Co., Ltd.) is added to 200 parts by weight of water together with 10 parts by weight of a partially decomposed starch (DE8, trade name “Paindex # 1”, manufactured by Matsutani Chemical Co., Ltd.) And dissolved by heating. After adjusting the pH to 6.0 by adding an appropriate amount of sodium hydroxide, 200 U cyclodextrin glucanotransferase (produced by Hayashibara Biochemical Laboratories) was added per 1 g of the partially decomposed starch and incubated at 40 ° C. for 24 hours. . Thereafter, the enzyme is inactivated by incubating at 100 ° C. for 20 minutes, followed by filtration with filter paper. Further, it is loaded onto a synthetic adsorbent (trade name “HP10”, manufactured by Mitsubishi Chemical Corporation) and adsorbed, The fractions eluted with 10 to 20% (w / w) ethanol aqueous solution were collected after washing with starch and the gallic saccharide used for the antimicrobial agent of the present invention. A transfer was obtained.
[0042]
When this was subjected to a conventional high performance liquid chromatography (HPLC) analysis, ultraviolet absorption was measured and a composition ratio (molar ratio) was determined, a gallic acid sugar transfer product having a saccharide portion having a glucose polymerization degree of 1 was obtained. 10%, 4% gallic acid glycosyl transfer product having a saccharide moiety having a glucose polymerization degree of 2 and 6% gallic acid glycosyl transfer product having a saccharide having a glucose polymerization degree of 6 or more, remaining unreacted gallic acid The acid acid was 80%.
[0043]
Example 1-2
An appropriate amount of sodium hydroxide was added to the reaction solution after the cyclodextrin glucanotransferase treatment obtained in Example 1-1 to adjust the pH to 4.5, and then 50 U of glucoamylase (product) per gram of the partially degraded starch. The name “Glucoteam” (manufactured by Nagase Seikagaku Corporation) was added and incubated at 40 ° C. for 20 hours. Subsequently, the enzyme was inactivated by incubating at 80 ° C. for 10 minutes, followed by filtration with filter paper as in Example 1-1, and further, a synthetic adsorbent (trade name “HP10”, Mitsubishi Chemical Corporation). The product was washed with water to elute the partially decomposed starch, and the fraction eluted with 20-30% (w / w) ethanol aqueous solution was collected and dried under reduced pressure. A gallic acid sugar transfer product used for the antimicrobial agent of the invention was obtained.
[0044]
When this was subjected to a conventional high performance liquid chromatography (HPLC) analysis, ultraviolet absorption was measured and a composition ratio (molar ratio) was determined, a gallic acid sugar transfer product having a saccharide portion having a glucose polymerization degree of 1 was obtained. 80%, 4% of the gallic acid glycosyl transfer product having a saccharide moiety having a glucose polymerization degree of 2 and 1% of the gallic acid glycosyl transfer product having a saccharide part having a glucose polymerization degree of 3 or more remained unreacted. The gallic acid was 15%.
[0045]
[Example 2]
<Preparation of gallic acid n-propyl sugar transfer product>
[0046]
Example 2-1
In accordance with the method of Example 1-1, 1 part by mass of n-propyl gallate (manufactured by Kanto Chemical Co., Ltd.) was changed to 10 parts by mass of a partially decomposed starch (DE8, trade name “Paindex # 1”, Matsutani). It was added to 200 parts by mass of water together with Chemical Co., Ltd. and dissolved by heating. After adjusting the pH to 6.0 by adding an appropriate amount of sodium hydroxide, 200 U cyclodextrin glucanotransferase (produced by Hayashibara Biochemical Laboratories) was added per 1 g of the partially decomposed starch and incubated at 40 ° C. for 24 hours. .
Thereafter, the enzyme is inactivated by incubating at 100 ° C. for 20 minutes, followed by filtration with filter paper. Further, it is loaded onto a synthetic adsorbent (trade name “HP10”, manufactured by Mitsubishi Chemical Corporation) and adsorbed, The fractions eluted with 35% to 50% (w / w) ethanol aqueous solution were collected by washing with ethanol, and the fraction eluted with ethanol aqueous solution was collected, dried under reduced pressure, and gallic acid n used in the antimicrobial agent of the present invention. A propyl sugar transfer product was obtained.
[0047]
This was subjected to conventional high performance liquid chromatography (HPLC) analysis, ultraviolet absorption was measured to determine the composition ratio (molar ratio), and n-propyl sugar gallate having a saccharide portion having a glucose polymerization degree of 1 N-propyl gallate having a saccharide portion having a sugar moiety having a sugar content of 14% and a glucose polymerization degree of 2%, and 9% of a n-propyl gallate having a sugar having a glucose polymerization degree of 3 or more is 9%. %, N-propyl gallate remaining unreacted was 67%.
[0048]
Example 2-2
In accordance with the method of Example 1-2, the reaction solution after the cyclodextrin glucanotransferase treatment obtained in Example 2-1 was adjusted to pH 4.5 by adding an appropriate amount of sodium hydroxide, and then starch. 50 U of glucoamylase (trade name “Glucoteam”, manufactured by Nagase Seikagaku Corporation) per 1 g of the partially decomposed product was added and incubated at 40 ° C. for 20 hours. Next, after incubating at 80 ° C. for 10 minutes to inactivate the enzyme, it was filtered with a filter paper as in Example 2-1, and further a synthetic adsorbent (trade name “HP10”, Mitsubishi Chemical Corporation). The product was adsorbed by loading and washed with water to elute the partially decomposed starch, and then the fraction eluted with 35-50% (w / w) aqueous ethanol solution was collected and dried under reduced pressure. A gallic acid n-propyl glycosyl transfer product used for the antimicrobial agent of the invention was obtained.
[0049]
This was subjected to conventional high performance liquid chromatography (HPLC) analysis, ultraviolet absorption was measured to determine the composition ratio (molar ratio), and n-propyl sugar gallate having a saccharide portion having a glucose polymerization degree of 1 N-propyl gallate having a saccharide portion having a saccharide moiety having a transfer rate of 76% and a glucose polymerization degree of 2 is 7%, and an n-propyl gallate transfer product having a saccharide portion having a glucose polymerization degree of 3 or more. 2%, n-propyl gallate remaining unreacted was 15%.
[0050]
[Example 3]
<Preparation of octyl gallate transfer product>
[0051]
Example 3-1
In accordance with the method of Example 1-1, 1 part by mass of gallic octyl (manufactured by Kanto Chemical Co., Ltd.) and 10 parts by mass of a partially decomposed starch (DE8, trade name “Paindex # 1”, Matsutani Chemical Co., Ltd.) The product was added to 200 parts by mass of water and dissolved by heating. After adjusting the pH to 6.0 by adding an appropriate amount of sodium hydroxide, 200 U cyclodextrin glucanotransferase (produced by Hayashibara Biochemical Laboratories) was added per 1 g of the partially decomposed starch and incubated at 40 ° C. for 24 hours. . Thereafter, the enzyme is inactivated by incubating at 100 ° C. for 20 minutes, followed by filtration with filter paper. Further, it is loaded onto a synthetic adsorbent (trade name “HP10”, manufactured by Mitsubishi Chemical Corporation) and adsorbed, The fractions eluted with 35 to 50% (w / w) ethanol aqueous solution were collected by washing with ethanol, and the fraction eluted with ethanol aqueous solution was collected, dried under reduced pressure, and octyl gallate used for the antimicrobial agent of the present invention. A glycosylated product was obtained.
[0052]
This was subjected to a conventional high performance liquid chromatography (HPLC) analysis, and ultraviolet absorption was measured to determine a composition ratio (molar ratio). As a result, an octyl gallate transfer product having a saccharide portion having a glucose polymerization degree of 1 was obtained. 14%, octyl gallate glyceryl transfer product having a saccharide portion having a glucose polymerization degree of 2 is 5%, octyl gallate transfer product having a saccharide having a glucose polymerization degree of 3 or more is 7%, and remains unreacted The remaining octyl gallate was 74%.
[0053]
Example 3-2
In accordance with the method of Example 1-2, the reaction liquid after the cyclodextrin glucanotransferase treatment obtained in Example 3-1 was adjusted to pH 4.5 by adding an appropriate amount of sodium hydroxide, and then starch. 50 U of glucoamylase (trade name “Glucoteam”, manufactured by Nagase Seikagaku Corporation) per 1 g of the partially decomposed product was added and incubated at 40 ° C. for 20 hours. Subsequently, after incubating at 80 ° C. for 10 minutes to inactivate the enzyme, it was filtered with a filter paper as in Example 3-1, and further a synthetic adsorbent (trade name “HP10”, Mitsubishi Chemical Corporation). The product was adsorbed by loading and washed with water to elute the partially decomposed starch, and then the fraction eluted with 35-50% (w / w) aqueous ethanol solution was collected and dried under reduced pressure. The octyl gallate transfer product used for the antimicrobial agent of the invention was obtained.
[0054]
This was subjected to a conventional high performance liquid chromatography (HPLC) analysis, and ultraviolet absorption was measured to determine a composition ratio (molar ratio). As a result, an octyl gallate transfer product having a saccharide portion having a glucose polymerization degree of 1 was obtained. Of octyl gallate having a saccharide portion with a glucose polymerization degree of 2 and 2%, 1% of an octyl gallate transferase having a saccharide portion with a glucose polymerization degree of 3 or more, unreacted The remaining octyl gallate was 21%.
[0055]
[Example 4]
<Preparation of caffeic acid sugar transfer product>
[0056]
Example 4-1
In accordance with the method of Example 1-1, 1 part by mass of caffeic acid (manufactured by Wako Pure Chemical Industries, Ltd.) was changed to 10 parts by mass of a partially decomposed starch (DE8, trade name “Paindex # 1”, Matsutani Chemical). The product was added to 200 parts by mass of water together with the product and dissolved by heating. After adjusting the pH to 6.0 by adding an appropriate amount of sodium hydroxide, 200 U cyclodextrin glucanotransferase (produced by Hayashibara Biochemical Laboratories) was added per 1 g of the partially decomposed starch and incubated at 40 ° C. for 24 hours. . Thereafter, the enzyme is inactivated by incubating at 100 ° C. for 20 minutes, followed by filtration with filter paper. Further, it is loaded onto a synthetic adsorbent (trade name “HP10”, manufactured by Mitsubishi Chemical Corporation) and adsorbed, After washing with starch to elute the partially decomposed starch, the fraction eluted with 35-50% (w / w) ethanol aqueous solution was collected, dried under reduced pressure, and caffeic acid sugar used for the antimicrobial agent of the present invention A transfer was obtained.
[0057]
When this was subjected to a conventional high performance liquid chromatography (HPLC) analysis, ultraviolet absorption was measured and a composition ratio (molar ratio) was obtained, a caffeic acid sugar transfer product having a saccharide portion having a glucose polymerization degree of 1 was obtained. 19%, 9% caffeic acid glycosyl transfer product having a sugar moiety having a glucose polymerization degree of 2, 9% caffeic acid glycosyl transfer product having a sugar having a glucose polymerization degree of 3 or more, and remaining unreacted cafe The acid was 65%.
[0058]
Example 4-2
In accordance with the method of Example 1-2, after adding the appropriate amount of sodium hydroxide to the reaction solution after the cyclodextrin glucanotransferase treatment obtained in Example 4-1, the starch was adjusted to pH 4.5. 50 U of glucoamylase (trade name “Glucoteam”, manufactured by Nagase Seikagaku Corporation) per 1 g of the partially decomposed product was added and incubated at 40 ° C. for 20 hours. Subsequently, after incubating at 80 ° C. for 10 minutes to inactivate the enzyme, it was filtered with a filter paper as in Example 4-1, and further a synthetic adsorbent (trade name “HP10”, Mitsubishi Chemical Corporation). The product was washed with water to elute the partially decomposed starch, and the fraction eluted with 20-30% (w / w) ethanol aqueous solution was collected and dried under reduced pressure. A caffeic acid sugar transfer product used for the antimicrobial agent of the invention was obtained.
[0059]
When this was subjected to a conventional high performance liquid chromatography (HPLC) analysis, ultraviolet absorption was measured and a composition ratio (molar ratio) was obtained, a caffeic acid sugar transfer product having a saccharide portion having a glucose polymerization degree of 1 was obtained. 78%, 3% caffeic acid glycosyl transfer product having a sugar moiety having a glucose polymerization degree of 2 and 1% caffeic acid glycosyl transfer product having a sugar part having a glucose polymerization degree of 3 or more remained unreacted. Caffeic acid was 18%.
[0060]
[Example 5]
<Preparation of glycosylated product of Terminaria berylica extract>
Momotamana Terminaria Berylica (Terminaria  bellicica) Was pulverized with a blender, 10 parts by mass of 50% (w / w) ethanol aqueous solution was added, and the mixture was stirred at 50 ° C. for 60 minutes. The filtrate was collected by filtration with filter paper, and the supernatant from which the solid components were removed by centrifugation was dried under reduced pressure to obtain a solid content. In accordance with the method of Example 1-1, 1 part by mass of this solid content was 200 parts by mass with 10 parts by mass of a partially decomposed starch (DE8, trade name “Paindex # 1”, manufactured by Matsutani Chemical Co., Ltd.). It was added to water and dissolved by heating. After adjusting the pH to 6.0 by adding an appropriate amount of sodium hydroxide, 200 U cyclodextrin glucanotransferase (produced by Hayashibara Biochemical Laboratories) was added per 1 g of the partially decomposed starch and incubated at 40 ° C. for 24 hours. . Then, after incubating at 100 ° C. for 20 minutes to inactivate the enzyme, according to the method of Example 1-2, an appropriate amount of sodium hydroxide was added to adjust the pH to 4.5, and then the starch partially decomposed product 50 U of glucoamylase (trade name “Glucoteam”, manufactured by Nagase Seikagaku Corporation) per 1 g was added and incubated at 40 ° C. for 20 hours. Next, the enzyme is inactivated by incubating at 80 ° C. for 10 minutes, and then filtered through a filter paper. The filtrate is collected, dried under reduced pressure, and sugar of the terminaria berylica extract used in the antimicrobial agent of the present invention. A transfer was obtained.
[0061]
The filtrate was subjected to a conventional high performance liquid chromatography (HPLC) method, and the content of the gallic acid sugar transfer product was measured. As a result, the gallic acid sugar transfer product as an active ingredient was 8 mg / g, gallic acid per solid content. Was 7 mg / g. In addition, a small amount of pentagalloylglucose sugar transfer product was detected.
[0062]
[Example 6]
<Glucose transfer product of indigo extract>
After crushing 1 part by mass of the above ground part of Polygonum Tincorium with a blender, 1.5 parts by mass of water was added and stirred with a mixer. The filtrate was collected by filtration with filter paper, heated at 121 ° C. for 10 minutes, centrifuged to remove solids, and the supernatant was collected. Next, the supernatant was dried under reduced pressure to obtain a solid content. In accordance with the method of Example 1-1, 200 parts by mass of starch partial decomposition product (DE8, trade name “Paindex # 1”, Matsutani Chemical Co., Ltd.) of 10 parts by mass with respect to 1 part by mass of the indigo extract. It was added to parts by weight of water and dissolved by heating. After adjusting the pH to 6.0 by adding an appropriate amount of sodium hydroxide, 200 U cyclodextrin glucanotransferase (produced by Hayashibara Biochemical Laboratories) was added per 1 g of the partially decomposed starch and incubated at 40 ° C. for 24 hours. . Thereafter, the enzyme is inactivated by incubating at 100 ° C. for 20 minutes, and then filtered through filter paper. The filtrate is collected, dried under reduced pressure, and the sugar of the Tade indigo extract used for the antimicrobial agent of the present invention. A transfer was obtained.
[0063]
The filtrate was subjected to a conventional high performance liquid chromatography (HPLC) method, and the content of caffeic acid glycosylated product was measured. As a result, the caffeic acid glycosylated product as an active ingredient was 5 mg / g, caffeic acid per solid content. Was included at a concentration of 12 mg / g.
[0064]
[Example 7]
<Oral cleaner>
According to a conventional method, a gallic acid sugar transfer product obtained by the method of Example 1-1 and an n-propyl sugar gallate obtained by the method of Example 2-1 with respect to the basic formulation of the oral cleaning agent shown below. Transfer product, octyl gallate transfer product obtained by the method of Example 3-1, caffeic acid transfer product obtained by the method of Example 4-1, sugar of terminaria berylica extract obtained by the method of Example 5 The transferred product or the sugar transfer product of the Tade indigo extract obtained by the method of Example 6 was blended to a concentration of 1% (w / w) to obtain a liquid oral cleanser.
[0065]
40 parts by mass of ethanol
15 parts by mass of glycerin
1 part by weight of polyoxyethylene hydrogenated castor oil
Saccharin appropriate amount
Perfume
Chlorhexidine appropriate amount
44 parts by weight of water
[0066]
The oral cleansing agent of this example exhibits antimicrobial properties against gram-negative bacteria, gram-positive bacteria, fungi, yeast, etc. in the oral cavity by being used in the same manner as normal mouthwashes. For example, Bacillus, Mycobacterium, Pseudomonas, Esseria, Staphylococcus, Streptococcus, Salmonella, Klebsiella, Proteus, Vibrio, Shigella, Helicobacter, Chlamydia, Aspergillus and Candida It is possible to effectively suppress the growth of microorganisms and effectively treat and prevent various diseases that cause their onset.
[0067]
[Example 8]
<Tablets>
In accordance with a conventional method, maltose powder was transferred to a gallic acid transfer product obtained by the method of Example 1-2, a gallic acid n-propyl transfer product obtained by the method of Example 2-2, and Example 3-2. Octyl gallate transfer product obtained by the method, caffeic acid transfer product obtained by the method of Example 4-2, transfer product of Terminaria berylica extract obtained by the method of Example 5, or the method of Example 6 After blending the sugar transfer product of Tade indigo extract obtained in 1 to a concentration of 1% (w / w), each tablet was tableted to a diameter of 1 cm and a thickness of 3.2 mm with a tableting machine. .5 g tablets were obtained.
[0068]
The antimicrobial agent of this example is dissolved in a living body rapidly by being used in the same manner as a normal tablet, suppresses the growth of microorganisms in the living body, and has excellent sustainability in the living body. It exhibits antimicrobial properties against Gram-negative bacteria, Gram-positive bacteria, fungi, yeasts and the like. For example, Bacillus, Mycobacterium, Pseudomonas, Esseria, Staphylococcus, Streptococcus, Salmonella, Klebsiella, Proteus, Vibrio, Shigella, Helicobacter, Chlamydia, Aspergillus, Candida It is possible to effectively treat and prevent diseases caused by other microorganisms.
[0069]
[Example 9]
<Toothpaste>
According to a conventional method, the gallic acid sugar transfer product obtained by the method of Example 1-1 and the n-propyl sugar gallate transfer obtained by the method of Example 2-1 with respect to the basic formulation of the toothpaste shown below. , Octyl gallate glyceryl transfer product obtained by the method of Example 3-1, caffeic acid sugar transfer product obtained by the method of Example 4-1, and sugar transfer of Terminaria berylica extract obtained by the method of Example 5 Or the sugar transfer product of the Tade indigo extract obtained by the method of Example 6 to a concentration of 1% (w / w), respectively, and then filled into an aluminum laminate tube by 100 g. An antimicrobial toothpaste was obtained.
[0070]
42 parts by weight of dibasic calcium phosphate
18 parts by mass of glycerin
Carrageenan 0.9 parts by mass
Sodium lauryl sulfate 1.1 parts by mass
Perfume
Saccharin appropriate amount
38 parts by weight of water
[0071]
The antibacterial toothpaste of the present example exhibits antimicrobial effects on gram-negative bacteria, gram-positive bacteria, fungi, yeast, etc. in the oral cavity by being used in the same manner as normal toothpastes. Therefore, Bacillus, Mycobacterium, Pseudomonas, Esseria, Staphylococcus, Streptococcus, Salmonella, Klebsiella, Proteus, Vibrio, Shigella, Helicobacter, Chlamydia, Aspergillus, Candida It is possible to effectively treat and prevent diseases caused by other microorganisms.
[0072]
[Example 10]
<Troche>
According to a conventional method, a gallic acid sugar transfer product obtained by the method of Example 1-1 and a gallic acid n-propyl sugar transfer obtained by the method of Example 2-1 with respect to the basic formulation of the lozenge shown below. , Octyl gallate glyceryl transfer product obtained by the method of Example 3-1, caffeic acid sugar transfer product obtained by the method of Example 4-1, and glycosylation of Terminaria berylica extract obtained by the method of Example 5 Or a sugar-transferred product of Tade indigo extract obtained by the method of Example 6 so as to have a concentration of 1% (w / w), respectively, and molded into a solid antimicrobial troche agent It was.
[0073]
35 parts by weight of maltose
34 parts by weight of starch
10 parts by mass of crystalline cellulose
Hydroxypropyl methylcellulose 10 parts by mass
Magnesium stearate 1 part by mass
[0074]
The antimicrobial lozenges of this example are dissolved in the oral cavity by regular use in the same way as normal lozenges, and have antimicrobial effects on gram negative bacteria, gram positive bacteria, fungi, yeast, etc. in the oral cavity. Indicates. Therefore, Bacillus, Mycobacterium, Pseudomonas, Esseria, Staphylococcus, Streptococcus, Salmonella, Klebsiella, Proteus, Vibrio, Shigella, Helicobacter, Chlamydia, Aspergillus, Candida It is possible to effectively treat and prevent diseases caused by other microorganisms.
[0075]
Example 11
<Liquid>
According to a conventional method, gallic acid sugar transfer product obtained by the method of Example 1-2 and purified gallic acid n-propyl sugar obtained by the method of Example 2-2 in purified water for injection containing 4% by mass of isoleucine Transfer product, octyl gallate transfer product obtained by the method of Example 3-2, caffeic acid transfer product obtained by the method of Example 4-2, sugar of terminaria berylica extract obtained by the method of Example 5 The transferred product, or the sugar transferred product of the Tade indigo extract obtained by the method of Example 6 is dissolved to a concentration of 1% (w / w) and then microfiltered and filled into a container to form a liquid antimicrobial A liquid having an effect was obtained.
[0076]
The liquid agent having an antimicrobial effect of this example is an injectable agent, a transmucosal agent, a nasal agent, an oral agent, and the like in the respiratory tract, nasal cavity, oral cavity, visual organ, digestive organ, urinary organ, genital organ, etc. Has antimicrobial effects on negative bacteria, gram positive bacteria, fungi, yeast, etc. Therefore, Bacillus, Mycobacterium, Pseudomonas, Esseria, Staphylococcus, Streptococcus, Salmonella, Klebsiella, Proteus, Vibrio, Shigella, Helicobacter, Chlamydia, Aspergillus, Candida It is possible to effectively treat and prevent diseases caused by other microorganisms.
[0077]
Example 12
<Ointment>
According to a conventional method, a gallic acid sugar transfer product obtained by the method of Example 1-1, a gallic acid n-propyl sugar transfer product obtained by the method of Example 2-1, and a method of Example 3-1. Octyl gallate transfer product, caffeic acid transfer product obtained by the method of Example 4-1, Terminaria berylica extract transfer product obtained by the method of Example 5, or the feed obtained by the method of Example 6 The sugar transfer product of indigo extract is dissolved in an appropriate amount of phosphate buffered saline (pH 7.2) so as to have a concentration of 1% (w / w), and 100 parts by mass of the aqueous solution is homogeneous with 49 parts by mass of anhydrous crystalline maltose. Then, the mixture was further kneaded with 450 parts by mass of white petrolatum to obtain an antimicrobial ointment.
[0078]
The antimicrobial ointment of this example exhibits antimicrobial effects on gram-negative bacteria, gram-positive bacteria, fungi, yeast, etc. in the skin, urinary organs, genital organs, etc., as a skin external preparation, transmucosal preparation and the like. Therefore, Bacillus, Mycobacterium, Pseudomonas, Esseria, Staphylococcus, Streptococcus, Salmonella, Klebsiella, Proteus, Vibrio, Shigella, Helicobacter, Chlamydia, Aspergillus, Candida It is possible to effectively treat and prevent diseases caused by other microorganisms.
[0079]
Example 13
<Eye drops>
According to a conventional method, the gallic acid sugar transfer product obtained by the method of Example 1-2 and the n-propyl sugar gallate transfer obtained by the method of Example 2-2 with respect to the basic formulation of the eye drops shown below. , Octyl gallate glyceryl transfer product obtained by the method of Example 3-2, caffeic acid saccharide transfer product obtained by the method of Example 4-2, and glyceryl transfer of Terminaria berylica extract obtained by the method of Example 5 Or a sugar-transfer product of Tade indigo extract obtained by the method of Example 6 was blended to a concentration of 1% (w / w) to obtain a liquid antimicrobial eye drop.
[0080]
20 parts by mass of trehalose (2 hydrates)
Sodium chloride 0.04 parts by mass
0.02 parts by mass of potassium chloride
Sodium dihydrogen phosphate 0.03 parts by mass
0.04 parts by mass of borax
Benzalkonium chloride 0.004 parts by mass
80 parts by mass of purified water
[0081]
The antimicrobial ophthalmic solution of this example exhibits antimicrobial effects on Gram-negative bacteria, Gram-positive bacteria, fungi, yeast, etc. in the visual instrument by being used in the same manner as ordinary eye drops. Therefore, Bacillus, Mycobacterium, Pseudomonas, Esseria, Staphylococcus, Streptococcus, Salmonella, Klebsiella, Proteus, Vibrio, Shigella, Helicobacter, Chlamydia, Aspergillus, Candida It is possible to effectively treat and prevent diseases caused by other microorganisms.
[0082]
【The invention's effect】
As described above, the antimicrobial agent of the present invention has low toxicity to the living body and is excellent in water solubility and excellent sustainability in the living body, so that it can be applied to living bodies such as injection or transdermal, transmucosal, nasal cavity or oral route. By applying it, it is possible to markedly suppress the growth of microorganisms that are undesirable for the living body. Therefore, microorganisms that have invaded the nasal cavity, oral cavity, esophagus, respiratory tract, visual organs, digestive organs, urinary organs, genital organs, etc. cause respiratory diseases such as pneumonia, arteriosclerosis, ischemic heart disease. Cardiovascular diseases such as conjunctivitis, digestive system diseases such as proctitis, urinary system diseases such as urethritis, accessory testicularitis, fallopian tubeitis, cervicitis, imminent premature birth, fallopian tube It is effective for the treatment and prevention of genital diseases such as inflammation, adhesion and obstruction, and food poisoning.
In particular, the oral preparation for antimicrobial use of the present invention, unlike oral preparations containing antibiotics as an active ingredient, causes the appearance of resistant bacteria, or produces a useful bacterial flora in the large intestine even after prolonged use. Since there is little risk of destruction, there is little risk of causing anorexia, vomiting, diarrhea, constipation, bloating, and causing side effects such as malaise and moisture.

Claims (1)

Bacillus genus, Esseria genus, Staphylococcus genus, Helicobacter genus, and Candida genus, comprising as an active ingredient a sugar transfer product in which a sugar having a glucose polymerization degree of 1 to 5 is added to caffeic acid through an α-glucoside bond anti-microbial agents that want to apply a microorganism belonging to the.