JP4599480B2

JP4599480B2 - Infectious immune function enhancer, metabolic function promoter, biological function deterioration prevention / amelioration agent, and functional foods containing these

Info

Publication number: JP4599480B2
Application number: JP2002057972A
Authority: JP
Inventors: 行雄柳本; 隆司渡辺
Original assignee: 柳本邦雄
Priority date: 2002-03-04
Filing date: 2002-03-04
Publication date: 2010-12-15
Anticipated expiration: 2022-03-04
Also published as: JP2003252787A

Description

【０００１】
【発明の属する技術分野】
本発明は、竹酢液及び/又は竹酢液を電気分解処理後に陽極から得られた酸性竹酢液（以下、電解竹酢酸性液と言う）を主成分とする皮膚組織新陳代謝機能促進剤、感染免疫機能強化剤、生体諸機能低下予防改善剤及びこれらを含有する機能食品に関し、詳しくは竹酢液及び/又は電解竹酢酸性液にシェア脂及びシェア脂含有成分を組み合わせて添加した組成物を有効成分皮膚組織新陳代謝機能促進剤、感染免疫機能強化剤、生体諸機能低下予防改善剤及びこれらを含有する機能食品に関する。
【０００２】
【従来の技術】
超高齢化社会現象が現実化した今日、加齢変化に随伴して生じ易くなる生活習慣病等の様々な疾患は医療費の増加のみならず医療制度そのものも崩壊寸前の状態であることは周知のところである。
一方、今後２１世紀の日常生活における高齢者介護、長期慢性疾患患者介護や在宅医療介護等に与える影響も大きく、重大な社会問題であり、また、若年・学童層の日常生活における不規則な食生活や不均衡栄養食摂取等から生じ易くなる免疫系機能低下症、神経系機能低下に伴う若年性痴呆症あるいはアトピー性皮膚炎やアレルギー性疾患などの発生・発症率の上昇も近年来顕著であり、介護関連問題と同様に重大な問題になりつつある。このような医療上の諸問題から、医療技術の更なる向上や介護制度の改善・改革と共に、年齢層を問わず食生活（介護食）の見直しと改善と改良が早急に要求されるところであう。
【０００３】
また、アトピー性皮膚炎や各臓器器官に生じる諸種アレルギー性疾患は、近年来、年齢層を問わず、これら疾患の発病・発症率は極めて高く、就中、乳幼期、学童期及び青年期におけるこれらの発病・発生率は顕著であり、日常の育児面や私生活面で多種多様の傷害作用因子を生み出しているのが現状である。一方、健康食品群の中にも、従来より、鶏卵、そば・麦粉、大豆（大豆製品）等は強力なアレルギー誘発食材あるいは商品群であることは周知のところであるが、最近では、これらに加えて米、コラーゲンや茸類等も高位置を占めるようになってきた。このような食生活面の観点からも、以前にも増して、抗アトピー剤や抗アレルギー剤の研究開発および商品化が盛んに行われているものの、即時的・即効的な抗アトピー剤や抗アレルギー剤は、何と言っても化学物質製剤やステロイド系製剤に依存しているが、完全治癒・治療に至っていない。他方、加齢変化と共に生体恒常性維持の低下、特に新陳代謝機能や免疫系機能が低下し、高年齢層での日和見感染症を伴ったアレルギー性皮膚疾患の治療対策は難問である。即ち、養護・介護施設や在宅における完全介護を必要とする寝たきり老人層の混合感染性・潰瘍性皮膚炎や褥蒼の治療対策は更に重大な社会問題となりつつある。多種に亘る治療剤の中で、注目されているのが、化学物質と天然物質を配合した製剤類である。例えば、褥蒼や潰瘍性皮膚疾患に対して有効であると言われている白糖・ポビドンヨード軟膏剤においても、ハロゲン族のヨウ素を主成分としているために多量投与及び長期投与による甲状腺機能への副作用、長期間保存下での効能安定性の低下や二層分離・水飴状による展延性劣化等の短所がみられ、今尚満足するに至る製剤とは言い難い（二神孝次郎、他．最新医学：１７；１３６９−１３７４，１９９２）。
【０００４】
これに対して、近年来竹酢液や木酢液がアトピー性皮膚炎やアレルギー性皮膚疾患の改善・治療に有効であることが、多くの研究報告に見られ、現在汎用されている多くのの竹酢液・木酢液商品は発癌性物質のフェナントレン誘導体（ベンズピレン）等やアルヒド誘導体（ホルムアルデヒド、アセトン）等の有害成分を多量に含有するタール分は殆ど除去されている（特開平５−９５６７９、特開平６−４９４５４、特開平９−９４２９１）。しかしながら、これら竹酢液・木酢液は、タール分除去製造工程中に、脱臭作用や殺菌作用を有する有効成分をも同時に相当量除去されている場合が多い。これに対して、本発明に供試した本発明者らによって開発された竹酢液及び/又は電解竹酢酸性液は、有害成分は限りなく除去され、且つ高い脱臭効能と殺菌効果が保持されていることが確認されている（特開２００１−３２３２８２）。更に、本竹酢液は、すでに世間で使用されているあるいは今後商品化されるであろう竹酢液類とは異なり、竹酢液そのものを多種食品に添加出来ること、即ち、食品添加物として、利用可能な製造法をすでに特許出願済みである（特開２０００−５３９７３）。
【０００５】
【発明が解決しようとする課題】
西アフリカから中央アフリカの乾燥地帯に自生するシェア木に実るプラムに似たシェア果実に含まれるシェア脂及びシェア脂の主要構成成分（リノール酸、リノレン酸等の不飽和脂肪酸、テルペノイド系、フィトステロイド系複素環式芳香化合物等）は、アフリカの強烈な日差しや乾燥から皮膚や肌を保護するために、古来よりシェアバター（ウイメンズゴールド）として伝承されており、アフリカの人々にとっては貴重な果実脂である。
そこで、本発明は、本発明者らによって開発された有害成分が除去され、且つ強い脱臭性能及び殺菌能を有する特定保険食品認定竹酢液及び/又はその電解竹酢酸性液の利点を更に生かすために、上記のシェア脂及び/又はシェア脂由来有効成分と配合して成る製剤は、２種組成物の相乗効果を期待し、全ての組成物が天然物由来から成るより優れたアトピー性皮膚炎治療改善剤やアレルギー性皮膚炎治療改善剤、潰瘍性皮膚炎治療改善剤あるいは熱傷や褥蒼等の損傷を受けた皮膚組織再生機能促進剤、日和見感染免疫機能強化剤、生体諸機能低下予防改善剤及びこれらを含有する機能食品を提供することを課題とする。
【０００６】
【課題を解決するための手段】
本発明者らは上記課題を解決するために、
（１）竹酢液、電解竹酢酸性液及び/又は電解竹酢アルカリ液の代表的日和見病原細菌のメチシリン耐性黄色ブドウ球菌（ＭＲＳＡ菌）、緑膿菌及び病原真菌のカンジダアルビカンス菌（カンジダ菌）に対する殺菌効果の比較検討並びに供試３種液の濃度依存的殺菌効果の検討
（２）強度熱傷マウスへの竹酢液及び/又は電解竹酢酸性液をシェア脂含有クリーム（以下、シェアクリームと言う）あるいはシェア脂含有ローション（以下、シェアローションと言う）に添加した配合剤の塗布あるいは塗抹による熱傷マウス創傷面におけるＭＲＳＡ菌とカンジダ菌混合感染防御機能促進効果の検討
（３）上記配合剤の塗布あるいは塗抹による熱傷マウス創傷面治癒促進効果の検討
（４）熱傷マウス皮下組織内への免疫担当細胞群の浸潤・遊走促進効果の検討並びに標的部位への皮下組織固有細胞群の増殖促進効果と結合組織主要蛋白成分（コラーゲン、エラスチン、ムコ蛋白質類）の産生促進効果の病理組織学的検討等を主眼として、鋭意検討した結果、本願の発明を完成するに至ったものである。
【０００７】
即ち、本願発明は、本発明者らによって開発され、しかも食品添加物である竹酢液（特開２０００−５３９７３）を独特の方法で処理した竹酢液及び/又は電解竹酢酸性液（特開２００１−３２３２８２）の有する特色（有害成分非含有、高度脱臭性能・殺菌性能成分含有）を生かすべく、アトピー性皮膚炎、アレルギー性皮膚炎や潰瘍性皮膚炎等の皮膚疾患に対する抗菌剤・殺菌剤並びにこれら皮膚疾患における皮膚組織再生機能促進剤を提供するものである。
【０００８】
即ち、本願の請求項１の発明は、竹酢液及び/又は竹酢液を電気分解処理後に陽極から得られた竹酢酸性液と、シェア木果実由来シェア脂及び/又はその抽出精製画分組成物とを必須成分とし、この竹酢酸性液の濃度が５〜２５％であるように配合されたことを特徴とする日和見菌混合感染症に対する抗菌及び/又は殺菌のための薬剤を提供する。
本願の請求項２の発明は、竹酢液及び/又は竹酢液を電気分解処理後に陽極から得られた竹酢酸性液と、シェア木果実由来シェア脂及び/又はその抽出精製画分組成物とを必須成分とし、この竹酢酸性液の濃度が５〜２５％であるように配合されたことを特徴とする皮膚組織新陳代謝機能促進剤を提供する。
本願の請求項３の発明は、竹酢液及び/又は竹酢液を電気分解処理後に陽極から得られた竹酢酸性液と、シェア木果実由来シェア脂及び/又はその抽出精製画分組成物を必須成分とし、この竹酢酸性液の濃度が５〜２５％であるように配合されたことを特徴とする損傷皮膚組織再生機能促進剤を提供する。
本願の請求項４の発明は、竹酢液及び/又は竹酢液を電気分解処理後に陽極から得られた竹酢酸性液と、シェア木果実由来シェア脂及び/又はその抽出精製画分組成物を必須成分とし、この竹酢酸性液の濃度が５〜２５％であるように配合されたことを特徴とする感染免疫機能強化剤を提供する。
【０００９】
【発明の実施の形態】
竹酢及び/又は電解竹酢酸性液は、イネ科タケ亜科植物群に属するモウソウ、マダケ、ハチク等３０種以上あり、本発明に関しては、そのいずれの品種でも実施可能であるが、本発明の課題・目的を達成するためには、本発明者らによって開発された製造方法（特開２００１−３２３２８２）に基づいて製造・製品化されたものを用いることが最も好ましい。即ち、植物から抽出された粗竹酢液に含まれる有害成分を除去後、電解処理することにより得られた酸化力のある物質を含有することを特徴とする竹酢液で、特に、植物を炭化処理した際に発生した煙を冷却して得られる粗竹酢液に含まれるタール成分の少なくとも一部を除去し、この除去処理を経た液を電解処理するものであることが最も好ましいが、タール成分などを除去していない粗竹酢液を、本願請求項に言う竹酢液として用いることもできる。また、本願発明にける竹酢液は、上記のタケ亜科植物群に属する竹を原材料としたもの以外に、他の木材を原材料とした木酢液を含む概念であり、所謂狭義の竹酢液と木酢液との両者を含む概念である。
このように、植物を炭化処理した際に発生した煙を冷却して得られる粗竹酢液を、必要に応じて濾過処理してタール成分等を除去し、処理された液のｐＨを３〜５の範囲に調整した後、電解処理することが望ましい。上記の電解処理が、粗竹酢液を隔膜を有する電解槽により、直流電解するものであり、陽極側で得られる竹酢電解酸性水を利用目的の原料として用いるか、粗竹酢液を無隔膜電解槽により、直流電解するものであり、得られた電解後竹酢液を利用目的の原料として用いるものである。上記の濾過処理としては、１．濾紙で濾過したのち、粉末状活性炭で処理したもの２．織布フィルターで濾過したのち、繊維状活性炭で処理したもの３．不織布フィルターで濾過したのち、ゼオライトで処理したもの４．竹炭で処理したもの５．木炭で処理したもの６．織布フィルターで濾過したのち、粒状活性炭で処理したもの等々、種々の方法を利用できる。
【００１０】
この製造法で製造・製品化された竹酢液は、現在広く市販されておらず、容易に入手することが困難であり、唯一の製造会社である立花バンブー株式会社の製品を本発明の実施に用いることができる。
他方、本発明に供試したシェア脂含有シェアクリーム及びシェアローションは、現在広く市販されておらず、アイ・エム・シー株式会社から入手可能である。
【００１１】
竹酢液及び/又は電解竹酢酸性液をシェア脂及びシェア脂主要構成成分に添加して成る組成物を有効成分とする本発明の感染免疫機能強化剤、新陳代謝機能促進剤及び生体諸機能低下予防改善剤の投与形態は、塗布・塗抹投与が一般的であるが、これに限定されるものではない。また、剤型も用途に応じて適宣選択でき、薬剤学的に許容される賦形剤、結合剤、崩壊剤、軟膏基剤、乳化剤、クリーム基剤、外用洗浄用基剤等を用いて、坐剤、軟膏剤、塗布剤、塗抹剤、乳状液剤及び/又は各種薬用ローション剤（薬用制汗剤、薬用抗口臭剤等）の医療用製剤及び/又は外用治療剤として供することができる。
また、本発明の感染免疫機能強化剤、新陳代謝機能促進剤及び生体諸機能低下予防改善剤は、それらの効能を失わない限り、他の薬効や生体諸機能賦活能を有する動・植物由来あるいは乳酸菌などの共生細菌由来成分を配合して成る医療用製剤として供することも可能である。例えば、本発明の感染免疫強化剤に各種のサイトカイン、抗酸化作用を有するビタミンや共生細菌由来成分を配合したり、更には天然物由来保湿成分を配合することにより、皮下組織成分再生の促進や血行の促進させ、皮膚や肌のシミ・シワ形成予防・消去を促進・亢進させる化粧品、化粧用乳状液及び/又は化粧用ローションとして供することができる。
【００１２】
【実施例】
次に、本願発明の目的と内容を理解させるべく実施例を挙げて説明するが、本発明は以下の実施例に限定されるものではない。
【００１３】
＜非電解竹酢液、電解竹酢酸性液及び電解竹酢アルカリ液の３種日和見病原菌に対する試験管内殺菌効果試験＞
試験に用いた非電解竹酢液、電解竹酢酸性液及び電解竹酢アルカリ液は、立花バンブー株式会社の製品を用いたが、これは、下記の製造方法に記載の方法と実質的に同様の方法で生産されたものである。
【００１４】
＜実施例１、２＞
非電解竹酢液、電解竹酢酸性液及び電解竹酢アルカリ液の製造方法
モウソウチクの乾留による竹炭の製法過程で採取された粗竹酢液を用いる。この粗竹酢液の組成は、有機物含有率５．５％、ホルマリン０．０３％、水分率９２．０％、酸含有率３．５％、ｐＨ２．４、比重１．０１８、暗黄褐色を呈する。そして、この粗竹酢液３００ｍｌに対して粒状活性炭３０ｇを添加し、約１時間攪拌し、その後２時間放置することで、透明な黄褐色の竹酢液（非電解竹酢液という）が得られた。得られた非電解竹酢液のｐＨは９．２であり、有機物含有率は０．３％に減少していた。この結果より、タール分が極端に減少し、ホルマリンも検出されなかつた。この処理液のｐＨを塩酸によりｐＨ３．８に調整した。そして、この調整液を、隔膜を有するバッチ式の電解槽を用いて電解処理を行った。電解条件は、電極問距離６０ｍｍ、電極白金−チタン、電極電解隔膜ポリエステル中性隔膜、電極面積７０×１２５ｍｍ、電解槽容積約５００ｃｃ、電解方法定電圧電解ＤＣ５５Ｖ、電解時間１０分であった。電解後に陽極から得られた竹酢液（電解竹酢酸性液という）のｐＨは２．４であり、陰極から得られた竹酢液（電解竹酢アルカリ液と言う）のｐＨは９．４であった。
ここで、非電解竹酢液を実施例１とし、陽極から得られた竹酢液（電解竹酢酸性液）を実施例２とする。
【００１５】
上記の非電解竹酢液（実施例１）、電解竹酢酸性液（実施例２）及び電解竹酢アルカリ液の３種日和見病原菌に対して、試験管内殺菌効果試験を行った。
（３種竹酢液の殺菌効果の比較）
非電解竹酢液、電解竹酢酸性液及び電解竹酢アルカリ液の各原液（ここでの原液とは、非希釈＝１００％濃度を表すものである）４．５ｍｌにあらかじめ分光光度計にて、菌液の濁度をＯＤ＝５４０ｎｍで調整した患者由来のＭＲＳＡ菌（７．５ｘ１０６ＣＦＵ／ｍｌ）、緑膿菌（５．８ｘ１０６ＣＦＵ／ｍｌ）あるいはカンジダ菌（８．５ｘ１０６ＣＦＵ／ｍｌ）の各菌液０．５ｍｌづつを添加する。［その際、理論的には３種竹酢液は９０％に希釈され、また各菌液の生菌単位（ＣＦＵ）は、ＭＲＳＡ菌＝３．８ｘ１０６ＣＦＵ／０．５ｍｌ＝３．８ｘ１０５ＣＦＵ／ｍｌ、緑膿菌では５．８ｘ１０６ＣＦＵ／０．５ｍｌ＝５．８ｘ１０５ＣＦＵ／ｍｌ、カンジダ菌では２．９ｘ１０６ＣＦＵ／０．５ｍｌ＝２．９ｘ１０５ＣＦＵ／ｍｌに希釈されることになる］。９種の混合試料液（５ｍｌ）を２５℃下、６０分間放置し、１０分毎に撹拌する。６０分後、９種の混合試料液をリン酸緩衝液（ＰＢＳ）にて１０倍階段希釈試し、その０．２ｍｌづつをプラスチックプレートに入れた後、ハートインフュージョン（ＨＩ）寒天培地（１５ｍｌ）を添加・混釈し、寒天が固まるまで放置する。これらＨＩ寒天培地プレートを３７℃下で１８時間培養した後、ＣＦＵを算定し、ｍｌ当たりのＣＦＵ値を求めた。その結果、表１に示すように、電解竹酢酸性液（即ち、実施例２）の供試３菌種日和見病原菌に対する殺菌効果（殺菌率）は非電解竹酢液及び電解竹酢アルカリ液のそれらよりも著しく、いずれの菌に対しても９３％以上の殺菌率であった。また、非電解竹酢液の３菌種に対する殺菌作用は、電解竹酢アルカリ液よりも相対的に強いものの、有意性は認められなかった。他方、３種竹酢液に対する供試３菌種の抵抗性は、ＭＲＳＡ菌（平均１６．６％）、カンジダ菌（平均１４．５％）及び緑膿菌（平均７．７％）の順であった。
【００１６】
【表１】

【００１７】
（３種竹酢液の濃度依存的殺菌効果）
上記の非電解竹酢液（実施例１）、電解竹酢酸性液（実施例２）及び電解竹酢アルカリ液の各原液をＰＢＳ液にて、５０％、２５％あるいは１０％に希釈した各試料液（４．５ｍｌ）にＭＲＳＡ菌液あるいはカンジダ菌液０．５ｍｌづつを加え、上記の比較実験と同様の方法を用いてＣＦＵを算定した。その結果、表２−ａと表２−ｂに示すように、３種供試竹酢液のＭＲＳＡ菌（表２−ａ）及びカンジダ菌（表２−ｂ）に対する殺菌効果（殺菌率）は、これら供試試料の濃度の上昇に伴って増加し、就中、５０％電解竹酢酸性液の殺菌率は、いずれの菌に対しても８３％以上であった。一方、カンジダ菌の方が、ＭＲＳＡ菌よりも３種竹酢液に対する感受性は高かった。
即ち、カンジダ菌の抵抗性はＭＲＳＡ菌よりも弱かった。これら結果（表１、２）からも、抗生剤に耐性を示す薬剤耐性菌の院内・養老施設内における水平的感染伝播は、重大な課題であり、汎用されている抗生剤は即効的抗菌・殺菌性を示すものの、生体に対する副作用・副反応の発生を避けることは、極めて困難である。これに対して、天然物由来抗菌・殺菌成分の耐性菌に対する効力は劣るものの、副作用を示すことが極めて低いことは、生理学的、細菌学的にも極めて興味深いものであろう。
【００１８】
【表２】

【００１９】
＜熱傷マウス創傷部位の作製法並びに熱傷創傷面積の測定＞
７週齢のＩＣＲ系雌マウス（平均体重：３２ｇ；日本クレア社製）の腹腔内にチオペンタール・ナトリウム麻酔剤（ラボナール、万有製薬社製）の０．１ｍｌ（２４ｍｇ／ｍｌ）を投与した後、電気バリカンにて背部の毛を剃り、電気ゴテ（２ｃｍｘ３ｃｍ、１００Ｖ/１００Ｗ）を除毛表面に３秒間当てて、熱傷創傷部位（６ｃｍ２＝１０％／体表面積）を作製した。この熱傷創はヒトでの生存限界である３度熱傷あるいは体表面積の１０％に相当する。熱傷直後及び熱傷後７日目毎、２８日間に亘り麻酔下のマウス背部創傷面にセロファン紙を当てて、セロファン紙上に創傷面をマーカーでトレースした後、画像解析装置（ＭｏｄｕｌａｔｏｒＳｙｓｔｅｍ，ＭＯＰ−ＡＭ０３、コントロン社製、ドイツ）にて創傷面積を計測した。
【００２０】
＜実施例３＞
シェアクリーム（アイ・エム・シー社製）３容に実施例２の電解竹酢酸性液（立花バンブー社製）１容を混合し作成した。この２５％電解竹酢酸性液・シェアクリーム配合剤（以下、２５％ＡＢＶ・ＳＣと略す）を実施例３とする。
尚、このＡＢＶ・ＳＣ配合剤の電解竹酢酸性液は濃度範囲の５〜２５％でも配合できる。また、シェアクリームの代わりにシェアローション（アイ・エム・シー社製）に上記濃度範囲の竹酢液、電解竹酢酸性液及び/又は電解竹酢アルカリ液を配合できる。
【００２１】
＜混合感染試験＞
混合感染実験系は、（１）配合クリーム非塗布対照群、（２）シェアクリーム（ＳＣ）単独塗布群、（３）電解竹酢酸性液（ＡＢＶ）、（４）２５％ＡＢＶ・ＳＣ塗布群の計３群（各群３匹宛）を準備し、以下の実験的混合感染実験を行った。
熱傷直後に熱傷創傷面の壊死表皮部分を剥離し、剥離面にＭＲＳＡ菌（５ｘ１０７ＣＦＵ）とカンジダ菌（５ｘ１０７ＣＦＵ）を等量づつ混合した混合菌液０．１ｍｌを滅菌綿棒で塗抹感染させ、感染後１日目と３日目に上記のシェアクリームあるいは２５％ＡＢＶ・ＳＣシェアクリームの各々０．５ｇづつを、滅菌爪楊枝で塗布・展延する。塗抹感染直後、７、１４、２１及び２８日目に滅菌ＰＢＳ液を浸した綿棒で感染創傷面を拭い取り、綿棒を０．５ｍｌＰＢＳ加試験管に移し、１０倍階段希釈した。希釈菌液中のＭＲＳＡ菌のＣＦＵ値はスタフィロコッカス１１０寒天培地にて、カンジダ菌のＣＦＵ値はカンジダＧＥ寒天培地にて各々算定し、熱傷創傷面積当たりのこれら感染菌のＣＦＵを求め、log ＣＦＵ／ｃｍ^２に換算した。その結果、シェアクリーム（ＳＣ）単独塗布感染群におけるＭＲＳＡ菌及びカンジダ菌の創傷面からのこれら感染菌の還元生菌数（log ＣＦＵ／ｃｍ^２) の減少推移を非塗布感染対照群のそれと比較した場合、表３の上段と中段に示すように、感染後７〜２８日間に亘って減少傾向は認められたものの、実験期間中のオーダー値は対照群のそれらと殆ど同値であり、両群間に有意差は認められなかった。これに対して、２５％電解竹酢酸性液（ＡＢＶ）単独塗布感染群での生菌単位の減少推移は著しく、感染１４日目以降では有意性がみられ、非塗布感染対照群間との有意差を検定 (Student's t-test 法) したところ、ＭＲＳＡ菌感染では、感染後１４日目に危険率５％以下、２１及び２８日目には危険率１％以下の有意差が（表：上段）、そしてカンジダ菌感染の場合には、感染１４日目及び２１日目に危険率５％以下、２８日目には危険率１％以下の有意差（表：中段）が得られた。一方、２５％電解竹酢酸性液添加シェアクリーム（ＡＢＶ・ＳＣ）塗布感染群でのこれら感染菌の減少推移は、電解竹酢酸性液単独塗布群のそれよりも極めて著しく、ＭＲＳＡ菌及びカンジダ菌のいずれに対しても、感染後１４日目の有意差は危険率１％以下、２１日目以降での両群間の有意差は、更に顕著であった（危険率０．１％以下）。他方、ＭＲＳＡ菌とカンジダ菌の混合感染における熱傷創傷面よりの減少推移を調べたところ、表の下段に示すように、２５％ＡＢＶ・ＳＣ塗布感染群の２１日目以降での混合感染菌の還元生菌数の減少は極めて顕著であった（危険率０．１％以下）。これら結果から、シェアクリームに電解竹酢酸性液を添加することにより、熱傷創傷面からの混合感染菌排除作用は相乗的に促進されることが明らかにされた。また、シェアクリームの硬度性と電解竹酢酸性液の殺菌作用を考慮した場合、電解竹酢酸性液の添加量の割合は、最終濃度が５〜２５％（クリーム濃度：９５〜７５％）になるように添加するのが適切であると考えられた。
また、シェアローション（ＳＬ）に非電解竹酢液（ＢＶ）を添加して作製されるＢＶ・ＳＬ液の場合でも、上記の濃度範囲（５〜２５％）が妥当であろうと考えられる。
【００２２】
【表３】

【００２３】
＜混合菌塗抹感染熱傷創傷面の皮下組織内浸潤細胞数と浸潤細胞分類＞
上記＜混合感染試験＞に記載した感染実験系４群（非塗布対照群及び塗布群３群）の塗抹感染後、７日目における創傷面の真皮層と皮下組織層間に１０％胎児牛血清加イーグルＭＥＭ培養液（ヘパリン：４単位/ｍｌ）５ｍｌを注入し、背部を十分揉みほぐした後、注入培養液をい注射器で回収、その１．０ｍｌをプラスチックマイクロプレート（直径３０ｍｍ；６ウエル）に移した。マイクロプレートを３７℃下、２時間保温（５％炭酸ガスインキュベーター）後、培養液を除去した。マイクロプレートをメタノール固定、ギムザ染色した後、ウエル上の総浸潤細胞数及び浸潤細胞の分類を顕微鏡下で算定した（図１参照）。その結果、表４：左側に示すように、電解竹酢酸性液（ＡＢＶ）単独塗布感染群の総浸潤細胞数は、非塗布感染対照群のそれよりも多少とも増加傾向が認められたものの、有意差はみられなかった。一方、塗布２群での総浸潤細胞数は、いずれも増加しており、非塗布感染対照群に対する有意差は、シェアクリーム（ＳＣ）単独塗布感染群では危険率５％以下そして２５％ＡＢＶ・ＳＣ塗布感染群では危険率１％以下であった。
他方、これら浸潤細胞の細胞分類を行ったところ、表：右側に示すように、非塗布感染対照群では、リンパ系細胞の比率が最高値（平均８９．５％）であり、マクロファージ等の網内系細胞は平均５．２％、上皮性及び繊維芽性細胞群は５．３％にすぎなかった。これに対して、電解竹酢酸性液単独塗布感染群では、マクロファージ様細胞は、約９％に増加していたものの、非塗布感染対照群の細胞分類比率と大差がなかった。
一方、シェアクリーム単独塗布感染群及び２５％ＡＢＶ・ＳＣ塗布感染群におけるマクロファージ様細胞比率は、著しく上昇し、それぞれ平均２３．４％と４６．８％を示し、２５％ＡＢＶ・ＳＣ塗布感染群における上皮性及び繊維芽性細胞群の上昇（平均１３．７％）も顕著であった（図１：参照）。
これら結果は、前述の＜混合感染試験＞における塗布感染２群の１４日目以降にみられた有意性のある混合感染菌排除促進作用の結果と一致していたことから、シェア脂及び/又はシェア脂主要構成成分は、免疫担当細胞群の第一細胞であるマクロファージの損傷部位や患部への遊走促進作用とマクロファージの機能亢進能（免疫強化能）を有することが、強く示唆された。更に、竹酢液は、糖質（３６３ｍｇ／ｇ）や食物繊維質（１７７ｍｇ／ｇ）を多量に含有していることから、これら成分も又、マクロファージに代表される免疫担当細胞群を活性化させている可能性も表４の結果から十分考えられる。
【００２４】
【表４】

【００２５】
＜混合菌感染熱傷創傷面皮下組織の病理組織学的検査＞
つぎに、上記の＜混合菌塗抹感染熱傷創傷面の皮下組織内浸潤細胞数と浸潤細胞分類＞で述べた推論を立証するために、以下の検討を行った。上記の４群中、電荷竹酢酸性液単独塗布群を除いた１４日目における平均創傷面積値に最も近いマウス創傷面の皮膚組織を切除し、定法に準じて処理した。即ち、脱水（アルコールシリーズ）、固定（１％グルタルアルデヒド、１％パラホルムアルデヒド）、パラフィン包埋した後、組織切片を作製した。更に、スライドグラスに処理した組織切片を付着・固定させた後、結合組織繊維性蛋白質を特異的に染め分けられるエラスチカファンギーソン染色液にて染色し、皮膚組織内の結合組織（膠原繊維、弾力繊維、コラーゲン・エラスチン等の繊維質）の再生・形成の程度を顕微鏡下で観察したところ、図２に示すよ感染免疫機能強化剤、新陳代謝機能促進剤、生体諸機能低下予防改善剤及うに、熱傷直後の皮下組織切片を、結合組織の状態、存在部位、形成量等についての組織学的観察が可能なエラスチカファンギーソン染色液で染色したところ、桃紅色に染色されている層は全く認められず、皮膚組織そのものが完全に壊死状態にあり、ヒトの３度熱傷に相当する熱傷の影響が真皮層のみならず皮下組織層にまで達していた。他方、熱傷後、１４日目における非塗布対照群の皮膚組織切片像では、上皮性（円形）細胞や繊維芽性細胞の真皮層内への浸潤が顕著であったが、桃紅色に染まった膠原繊維や弾力繊維（コラーゲン、エラスチン等）は極めて少なく、表皮層真皮層の再生は殆ど観察されなかった。これに対して、シェアクリーム（ＳＣ）単独塗布群での、熱傷後１４日目における皮膚組織切片を観察したところ、表皮層の再生は、殆ど完了しており、鮮やかな桃紅色に染色された部位は、表皮層から皮下組織層にまで認められ、繊維芽性細胞や上皮性細胞も殆どみられなかったが、染色された皮下組織層は、網目状が目立ち、皮下組織の完全なる再生には至ってはいなかった。一方、２５％ＡＢＶ・ＳＣ塗布群での皮膚組織切片を観察したところ、真皮層から皮下組織層に至るまで、全面濃桃紅色に染色されており、しかも染色された皮下組織層の緻密性は、ＳＣ単独塗布群のそれよりも極めて顕著であった。これら結果から、シェアクリームに電解竹酢酸性液を添加することにより、熱傷創傷面の日和見病原菌排除促進効果のみならず、竹酢液の構成成分である糖質、食物繊維質更には蛋白質（３６ｍｇ／ｇ）によって、上皮細胞や繊維芽細胞の諸機能が亢進されと同時にシェア脂の主要構成成分も又結合組織の再生促進、結合組織成分の産生促進、ひいては生体組織の新陳代謝機能が相乗的に促進されるであろうことが、病理組織学的観察からも確認された。
【００２６】
＜混合菌塗抹感染熱傷創傷面積の縮小・治癒推移＞
混合菌塗抹感染創傷面塗布実験系は、＜混合感染試験＞に記載したのと同様の３群のマウス（各群１０匹宛）を準備し、熱傷直後、熱傷後４、１４、２１及び２８日目の創傷面積を実施例２の方法で計測した。その結果、表５に示すように、熱傷直後の創傷面積を計測したところ、６．５３ ± ０．４１ cm 2 であり、電気ゴテの表面積 (６．０cm²) とほぼ近似の創傷面積が作製された。このような熱傷創傷面を用いて実験を行ったところ、電解竹酢酸性液（ＡＢＶ）単独塗布感染群における熱傷後７、１４、２１及び２８日目の創傷面積は、各々平均６．０６、４．９９、２．１６及び１．０１ｃｍ2であり、日数の経過と共に徐々に縮小し、非塗布感染対照群における７〜２８日目の平均創傷面積よりも低い示したものの、有意差は２８日目のみに観察されたに過ぎなかった（危険率５％以下）。これに対して、シェアクリーム（ＳＣ）単独塗布感染群では、熱傷後７日目で有意差（危険率５％以下、面積：５．４６±０．４９cm²)が認められ、１４日目以降では、更に高い有意差（危険率１％以下）を示した。一方、２５％ＡＢＶ・ＳＣ塗布感染群における創傷面積の縮小（治癒）は、前２群よりも著しく促進され、熱傷後７日目において、極めて高い有意性（危険率１％以下）のある創傷面積の縮小（４．８１±０．２９cm²)が観察され、１４日目以降での危険率はいずれも０．１％であった（図３参照）。
これらの結果を総括した場合、シェアクリームに電解竹酢酸性液を添加することにより、電解竹酢酸性液の混合感染菌排除・殺菌作用によって、シェアクリームが有する本来の皮膚炎・皮膚疾患治癒促進効果が感染熱傷創傷面治癒に対しても相乗的且つ効率よく発揮されたものであろうことが十分示唆された。
【００２７】
【表５】

【００２８】
＜毒性試験＞
非電解竹酢液、電解竹酢酸性液、シェアクリーム、シェアローション及びこれら竹酢液添加シェアクリーム及び/又はシェアローションの健康者（４０名）に対する毒性試験、即ち皮膚一次刺激性試験、光感作性（アレルギー）試験、光毒性試験を行ったところ、いずれの試験においても陰性であった。他方、ＩＣＲマウスを用いての非電解竹酢液及び電解竹酢酸性液の連続５回経口投与（１回/日）による毒性試験においても、毒性反応は観察されなかった（図表：省略）。
【００２９】
＜実施例４＞
褥蒼用・熱傷用湿布剤及び/又は軟膏剤
１０ｇのシェアクリーム（アイ・エム・シー社製）に下記表６［湿布剤］の電解竹酢酸性液、米由来糖セラミド及びマルチビタミン（Ａ，Ｅ，Ｃ）を混和し、更にシェアクリーム９０ｇを加えてシェアクリームの全含有量１００ｇの褥蒼・熱傷創傷面感染菌排除用及び創傷面治癒促進用湿布剤を製した。
同様に、１０ｇのシェアクリーム（アイ・エム・シー社製）に下記表６［軟膏剤］の電解竹酢酸性液、米由来糖セラミド及びマルチビタミン（Ａ，Ｅ，Ｃ）を混和し、さらに、白色ワセリン１００ｇを加えて褥蒼・熱傷創傷面感染菌排除用及び創傷面治癒促進用軟膏剤を製した。
【００３０】
【表６】
［湿布剤］
シェアクリーム１００ｇ
電解竹酢酸性液２５ｍｌ
米由来糖セラミド１００ｍｇ
マルチビタミン（Ａ，Ｅ，Ｃ）５００ｍｇ
白色ワセリン０ｇ
［軟膏剤］
シェアクリーム１０ｇ
電解竹酢酸性液２５ｍｌ
米由来糖セラミド１００ｍｇ
マルチビタミン（Ａ，Ｅ，Ｃ）５００ｍｇ
白色ワセリン１００ｇ
【００３１】
＜実施例５＞
１０ｍｌのシェアローション（アイ・エム・シー社製）に下記表７の成分を混合し、更にシェアローション９０ｍｌを加えてシミ、シワ形成予防・消去促進薬用・化粧用ローションを製した。
【００３２】
【表７】
［薬用・化粧用ローション］
シェアローション１００ｍｌ
非電解竹酢液１０ｍｌ
米由来糖セラミド５０ｍｇ
マルチビタミン（Ａ，Ｅ，Ｃ）１００ｍｇ
【００３３】
＜実施例６＞
非電解竹酢液、難溶性オリゴ糖及びマルチビタミンに、適量のブドウ糖液及び香料を加えて、滅菌精製水にて、全量１００ｍｌの新陳代謝機能促進用及び生体諸機能低下予防改善用の飲料（機能食品）を製した。
この機能食品（１００ｍｌ中含有成分量）の下記に示す。
【００３４】
【表８】
［機能食品］
非電解竹酢液１０ｍｌ
難溶性オリゴ糖２ｇ
マルチビタミン３００ｍｇ
【００３５】
【発明の効果】
本願発明は、竹酢酸性液と、シェア木果実由来シェア脂及び/又はその抽出精製画分組成物とを必須成分とした薬剤を提供し得たものであり、請求項１の発明は、日和見菌混合感染症に対する抗菌及び/又は殺菌効果のため薬剤を提供し得たものであり、請求項２の発明は、皮膚組織新陳代謝機能促進剤を提供し得たものであり、請求項３の発明は、損傷皮膚組織再生機能促進剤を提供し得たものであり、請求項４の発明は、感染免疫機能強化剤を提供し得たものである。
【図面の簡単な説明】
【図１】本願実施例に係る熱傷・混合感染後７日目における熱傷創傷面皮下組織内浸潤細胞像である。
【図２】本願実施例に係る熱傷・混合感染後１４日目における熱傷創傷面皮下組織切片像である。
【図３】本願実施例に係る熱傷・混合感染後２１日目における熱傷創傷面の縮小及び治療像である。[0001]
BACKGROUND OF THE INVENTION
The present invention is a skin tissue metabolic function promoter comprising, as a main component, an acidic bamboo vinegar solution obtained from an anode after electrolysis of bamboo vinegar solution and / or bamboo vinegar solution (hereinafter referred to as electrolytic bamboo acetic acid solution), Infectious immunity function enhancer, biological function deterioration prevention and improvement agent, and functional foods containing these, in particular, a composition comprising a combination of bamboo vinegar solution and / or electrolytic bamboo acetic acid solution combined with shear fat and shear fat-containing components The present invention relates to an active ingredient skin tissue metabolism function promoter, an infectious immune function enhancer, a biological function deterioration preventive / ameliorating agent, and a functional food containing these.
[0002]
[Prior art]
Now that the super-aging social phenomenon has become a reality, it is well known that various diseases such as lifestyle-related diseases that are likely to occur with aging changes are not just an increase in medical expenses, but the medical system itself is on the verge of collapse. It is the place.
On the other hand, it will have a significant impact on elderly care, long-term chronic care and long-term care for the 21st century, and is a serious social problem. Increasing incidence and incidence of immunological dysfunction, juvenile dementia, atopic dermatitis, and allergic diseases associated with decreased nervous system function, which are likely to occur due to daily life and unbalanced nutrition Yes, it is becoming a serious problem as well as nursing-related problems. Because of these medical problems, along with further improvement of medical technology and improvement / reform of the nursing care system, a review, improvement and improvement of dietary habits (care food) are urgently required regardless of age group. .
[0003]
In addition, atopic dermatitis and various allergic diseases that occur in various organs have been extremely high in recent years regardless of age group, especially during childhood, schoolchildhood and adolescence. The incidence and incidence of these in Japan are remarkable, and they currently produce a wide variety of injury-causing factors in terms of daily childcare and personal life. On the other hand, it is well known in the health food group that chicken eggs, buckwheat flour, soybeans (soybean products), etc. are strong allergy-inducing foods or product groups. Rice, collagen, and moss have come to occupy high positions. From the viewpoint of eating habits as well, although research and development and commercialization of anti-atopic and anti-allergic agents are more active than before, immediate and immediate anti-atopic agents and Allergic drugs depend on chemicals and steroids, but have not yet been completely cured or treated. On the other hand, the treatment of allergic skin diseases associated with opportunistic infections in the elderly is a difficult problem because the maintenance of living homeostasis, particularly the metabolic function and immune system function, declines with age. In other words, treatment measures for mixed infectious / ulcerative dermatitis and hemorrhoids in bedridden elderly people who need complete care at nursing homes or nursing homes or at home are becoming more serious social problems. Among a wide variety of therapeutic agents, attention is given to preparations in which chemical substances and natural substances are blended. For example, sucrose and povidone iodine ointment, which is said to be effective against epilepsy and ulcerative skin diseases, also have side effects on thyroid function due to high-dose and long-term administration because it mainly contains halogen group iodine. However, it is difficult to say that it is still a satisfactory formulation due to the decrease in efficacy stability under long-term storage and the deterioration of spreadability due to bilayer separation and varicella (Kojiro Futagami et al. : 17; 1369-1374, 1992).
[0004]
On the other hand, in recent years, many research reports have shown that bamboo vinegar and wood vinegar are effective for the improvement and treatment of atopic dermatitis and allergic skin diseases. Bamboo vinegar liquid and wood vinegar liquid products are almost free of tars containing a large amount of harmful components such as carcinogenic phenanthrene derivatives (benzpyrene) and alkyd derivatives (formaldehyde, acetone) (Japanese Patent Laid-Open No. 5-95679, JP-A-6-49454, JP-A-9-94291). However, in these bamboo vinegar and wood vinegar, a considerable amount of active ingredients having a deodorizing action and a bactericidal action is often removed at the same time during the tar removal process. On the other hand, the bamboo vinegar and / or electrolytic bamboo acetic acid solution developed by the present inventors who have been used in the present invention has no harmful components removed and retains a high deodorizing effect and bactericidal effect. (Japanese Patent Laid-Open No. 2001-323282). Furthermore, the bamboo vinegar is different from bamboo vinegar that has already been used in the world or will be commercialized in the future, and the bamboo vinegar itself can be added to various foods, that is, as a food additive. A patent application has already been filed for an available production method (Japanese Patent Laid-Open No. 2000-53973).
[0005]
[Problems to be solved by the invention]
Shear fat and main constituents of shear fat (saturated fatty acids such as linoleic acid, linolenic acid, terpenoids, phytosteroids) contained in fruit that resembles plums that grow on a share tree that grows naturally in dry areas from West Africa to Central Africa Heterocyclic aromatic compounds have been handed down as shea butter (women's gold) since ancient times to protect the skin and skin from the intense sunlight and dryness of Africa. is there.
Therefore, the present invention further takes advantage of the special insurance food certified bamboo vinegar liquid and / or its electrolytic bamboo acetic acid liquid that has been developed by the present inventors and has removed harmful components and has strong deodorizing performance and bactericidal ability. Therefore, a preparation comprising the above-mentioned shear fat and / or an active ingredient derived from shear fat is expected to have a synergistic effect of two kinds of compositions, and a superior atopic skin in which all compositions are derived from natural products. Remedies for inflammation treatment, treatment for allergic dermatitis, treatment for ulcerative dermatitis, skin tissue regeneration function damaged by burns, wrinkles, etc., opportunistic infectious immune function enhancement, prevention of deterioration of various biological functions It is an object to provide an improving agent and a functional food containing these.
[0006]
[Means for Solving the Problems]
In order to solve the above problems, the present inventors have
(1) Representative opportunistic pathogenic bacteria of methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa and pathogenic fungi Candida albicans (Candida) ) And the concentration-dependent bactericidal effect of the three test liquids
(2) A compounding agent obtained by adding bamboo vinegar solution and / or electrolytic bamboo acetic acid solution to severe burn mice to shear fat-containing cream (hereinafter referred to as shear cream) or shear fat-containing lotion (hereinafter referred to as shear lotion) Of the protective effect of MRSA and Candida mixed infection protection on the wound surface of burned mice by applying or smearing
(3) Examination of healing effect on wound surface of burned mice by applying or smearing the above combination
(4) Invasion / migration promotion effect of immunocompetent cells in burned mouse subcutaneous tissue, growth promotion effect of subcutaneous tissue specific cell group on target site and connective tissue major protein components (collagen, elastin, mucoproteins) As a result of intensive studies focusing on the histopathological examination of the production promoting effect), the present invention has been completed.
[0007]
That is, the present invention is a bamboo vinegar solution and / or an electrolytic bamboo acetic acid solution (specialized) obtained by treating the bamboo vinegar solution (Japanese Patent Laid-Open No. 2000-53973), which is a food additive, by a unique method. In order to take advantage of the characteristics (containing no harmful components, high deodorizing performance / bactericidal performance components) of the open 2001-323282), antibacterial agents / sterilization against skin diseases such as atopic dermatitis, allergic dermatitis and ulcerative dermatitis The present invention provides an agent and a skin tissue regeneration function promoter for these skin diseases.
[0008]
That is, the invention of claim 1 of the present application is the bamboo vinegar solution and / or bamboo acetate solution obtained from the anode after the electrolysis treatment, the share fruit-derived share fat and / or the extract and purified fraction thereof. An antibacterial and / or bactericidal agent against opportunistic mixed infections, characterized in that the composition is an essential component and is formulated so that the concentration of this bamboo acetate solution is 5 to 25% For Provide drugs.
Invention of Claim 2 of this application is the bamboo acetic acid liquid obtained from the anode after electrolyzing bamboo vinegar liquid and / or bamboo vinegar liquid, the share fat derived from a share tree fruit, and / or its extraction refinement | purification fraction composition Is provided as an essential component, and a skin tissue metabolic function promoter is provided, wherein the bamboo acetic acid solution has a concentration of 5 to 25%.
The invention of claim 3 of the present application is a bamboo acetic acid solution obtained from an anode after electrolysis of bamboo vinegar and / or bamboo vinegar, and a share fat derived from a share tree fruit and / or an extract / purified fraction composition thereof. Is provided as an essential component, and a damaged skin tissue regeneration function promoter characterized by being formulated so that the concentration of this bamboo acetate solution is 5 to 25% is provided.
Invention of Claim 4 of this application is the bamboo acetic acid liquid obtained from the anode after electrolyzing the bamboo vinegar liquid and / or the bamboo vinegar liquid, the share fat derived from the share tree fruit and / or the extract purified fraction composition thereof Is provided as an essential component, and an infectious immunity function-enhancing agent characterized in that the concentration of this bamboo acetic acid solution is 5 to 25% is provided.
[0009]
DETAILED DESCRIPTION OF THE INVENTION
There are more than 30 kinds of bamboo vinegar and / or electrolytic bamboo acetic acid solution such as Moso, Mushroom, Hachiku, etc. belonging to the family Gramineae, Bambooaceae, and in the present invention, any of the varieties can be implemented. In order to achieve the above-mentioned problems / objects, it is most preferable to use a product manufactured and commercialized based on a manufacturing method developed by the present inventors (Japanese Patent Laid-Open No. 2001-323282). That is, a bamboo vinegar liquid characterized by containing an oxidative substance obtained by electrolytic treatment after removing harmful components contained in a crude bamboo vinegar liquid extracted from a plant. It is most preferable to remove at least a part of the tar component contained in the crude bamboo vinegar obtained by cooling the smoke generated when carbonized, and to electrolyze the liquid that has undergone this removal treatment, A crude bamboo vinegar solution from which tar components and the like have not been removed can also be used as the bamboo vinegar solution described in the claims of this application. In addition, the bamboo vinegar in the present invention is a concept including a wood vinegar made from other wood, in addition to those made from bamboo belonging to the above-mentioned bamboo subfamily group. It is a concept that includes both vinegar and wood vinegar.
Thus, the crude bamboo vinegar obtained by cooling the smoke generated when carbonizing the plant is filtered as necessary to remove tar components, etc., and the pH of the treated liquid is adjusted to 3 to 3. It is desirable to perform electrolytic treatment after adjusting to the range of 5. The above electrolytic treatment is to subject the crude bamboo vinegar solution to direct current electrolysis in an electrolytic bath having a diaphragm. Either the bamboo vinegar electrolytic acid water obtained on the anode side is used as a raw material for utilization, or the crude bamboo vinegar solution is not used. In the diaphragm electrolytic cell, direct current electrolysis is performed, and the obtained post-electrolysis bamboo vinegar solution is used as a raw material for utilization. As said filtration process, 1. 1. Filtered with filter paper and then treated with powdered activated carbon 2. Filtered with a woven fabric filter and then treated with fibrous activated carbon. 3. Filtered with a nonwoven filter and then treated with zeolite. 4. Treated with bamboo charcoal Treated with charcoal 6. Various methods can be used, such as those that are filtered through a woven fabric filter and then treated with granular activated carbon.
[0010]
Bamboo vinegar produced and commercialized by this production method is not widely available at present and is difficult to obtain easily. The product of Tachibana Bamboo Co., Ltd., the only manufacturer, is used in the present invention. Can be used.
On the other hand, the shear fat-containing shear cream and shear lotion used in the present invention are not widely marketed at present and are available from IBM Corporation.
[0011]
Infectious immune function enhancer of the present invention, metabolic function promoter and decreased biological functions, comprising as an active ingredient a composition comprising bamboo vinegar and / or electrolytic bamboo acetic acid as a main component of shear fat and shear fat The administration form of the preventive / ameliorating agent is generally applied or smeared, but is not limited thereto. In addition, the dosage form can be appropriately selected according to the use, and using pharmaceutically acceptable excipients, binders, disintegrants, ointment bases, emulsifiers, cream bases, external cleaning bases, etc. Suppositories, ointments, coating agents, smears, emulsions and / or various medicinal lotions (medicine antiperspirants, medicinal anti-bad breath agents, etc.) and can be used as a medical preparation and / or an external therapeutic agent.
In addition, the infectious immune function enhancer, metabolic function promoter, and biological function deterioration prevention / amelioration agent of the present invention have other medicinal and biological function-activating activities or lactic acid bacteria, as long as they do not lose their efficacy. It is also possible to provide a medical preparation comprising a symbiotic bacterium-derived component. For example, various infectious immune enhancers of the present invention can be combined with various cytokines, antioxidant vitamins and symbiotic bacteria-derived components, and further with natural product-derived moisturizing components to promote the regeneration of subcutaneous tissue components. It can be used as cosmetics, cosmetic emulsions and / or cosmetic lotions that promote blood circulation and promote / enhance the prevention and elimination of skin and skin spots and wrinkles.
[0012]
【Example】
Next, in order to make the purpose and contents of the present invention understood, examples will be described, but the present invention is not limited to the following examples.
[0013]
<Test tube sterilization effect test against non-electrolytic bamboo vinegar solution, electrolytic bamboo acetic acid solution and electrolytic bamboo vinegar alkaline solution against opportunistic pathogens>
The non-electrolytic bamboo vinegar solution, the electrolytic bamboo acetic acid solution, and the electrolytic bamboo vinegar alkaline solution used in the test used products of Tachibana Bamboo Co., Ltd., which is substantially the same as the method described in the following production method. It was produced by the method of.
[0014]
<Examples 1 and 2>
Method for producing non-electrolytic bamboo vinegar, electrolytic bamboo acetate, and electrolytic bamboo vinegar alkaline solution
The crude bamboo vinegar liquid collected in the process of making bamboo charcoal by dry distillation of moso bamboo is used. The composition of this crude bamboo vinegar is: organic content 5.5%, formalin 0.03%, moisture content 92.0%, acid content 3.5%, pH 2.4, specific gravity 1.018, dark yellow Shows brown color. Then, 30 g of granular activated carbon is added to 300 ml of this crude bamboo vinegar, stirred for about 1 hour, and then left for 2 hours to obtain a transparent tan bamboo vinegar (referred to as non-electrolytic bamboo vinegar). It was. The obtained non-electrolytic bamboo vinegar had a pH of 9.2, and the organic matter content was reduced to 0.3%. From this result, the tar content was extremely reduced, and formalin was not detected. The pH of this treatment solution was adjusted to pH 3.8 with hydrochloric acid. And this electrolytic solution was electrolyzed using the batch type electrolytic cell which has a diaphragm. The electrolysis conditions were an electrode distance of 60 mm, an electrode platinum-titanium, an electrode electrolytic diaphragm, a polyester neutral diaphragm, an electrode area of 70 × 125 mm, an electrolytic cell volume of about 500 cc, an electrolysis method constant voltage electrolysis of DC 55 V, and an electrolysis time of 10 minutes. The pH of bamboo vinegar obtained from the anode after electrolysis (referred to as electrolytic bamboo acetic acid) is 2.4, and the pH of bamboo vinegar obtained from the cathode (referred to as electrolytic bamboo vinegar alkaline solution) is 9.4. Met.
Here, the non-electrolytic bamboo vinegar solution is referred to as Example 1, and the bamboo vinegar solution (electrolytic bamboo acetate-based solution) obtained from the anode is referred to as Example 2.
[0015]
An in-vitro bactericidal effect test was performed on the above three types of opportunistic pathogens, the non-electrolytic bamboo vinegar solution (Example 1), the electrolytic bamboo acetic acid solution (Example 2), and the electrolytic bamboo vinegar alkaline solution.
(Comparison of bactericidal effect of 3 kinds of bamboo vinegar)
Each stock solution of non-electrolytic bamboo vinegar solution, electrolytic bamboo acetic acid solution and electrolytic bamboo vinegar alkaline solution (the undiluted solution here represents undiluted = 100% concentration) in 4.5 ml with a spectrophotometer in advance MRSA bacteria (7.5 × 10 6 CFU / ml), Pseudomonas aeruginosa (5.8 × 10 6 CFU / ml), Candida (8.5 × 10 6 CFU / ml) derived from a patient whose turbidity was adjusted to OD = 540 nm Add 0.5 ml each. [In this case, the three kinds of bamboo vinegar are theoretically diluted to 90%, and the viable unit (CFU) of each bacterial liquid is MRSA bacteria = 3.8 × 10 6 CFU / 0.5 ml = 3.8 × 10 5 CFU / ml, Pseudomonas aeruginosa is diluted to 5.8 × 10 6 CFU / 0.5 ml = 5.8 × 10 5 CFU / ml, and Candida is diluted to 2.9 × 10 6 CFU / 0.5 ml = 2.9 × 10 5 CFU / ml]. Nine kinds of mixed sample liquids (5 ml) are allowed to stand at 25 ° C. for 60 minutes and stirred every 10 minutes. Sixty minutes later, nine mixed sample solutions were diluted 10-fold with phosphate buffered saline (PBS), and 0.2 ml of each was placed on a plastic plate, and then Heart Infusion (HI) agar medium (15 ml). Add and pour and leave until the agar has hardened. After culturing these HI agar plates at 37 ° C. for 18 hours, the CFU was calculated and the CFU value per ml was determined. As a result, as shown in Table 1, the bactericidal effect (bactericidal rate) of the electrolytic bamboo acetate-based liquid (ie, Example 2) against the test 3 bacterial species opportunistic pathogens of the non-electrolytic bamboo vinegar and the electrolytic bamboo vinegar alkaline liquid The sterilization rate was 93% or more for all the bacteria. Moreover, although the bactericidal action with respect to three microbe species of nonelectrolytic bamboo vinegar liquid is relatively stronger than electrolytic bamboo vinegar alkaline liquid, the significance was not recognized. On the other hand, the resistance of the three test species to the three kinds of bamboo vinegar was in the order of MRSA (average 16.6%), Candida (average 14.5%) and Pseudomonas aeruginosa (average 7.7%). Met.
[0016]
[Table 1]

[0017]
(Concentration-dependent bactericidal effect of 3 kinds of bamboo vinegar)
Each undiluted solution of the above non-electrolytic bamboo vinegar solution (Example 1), electrolytic bamboo acetic acid solution (Example 2) and electrolytic bamboo vinegar alkaline solution was diluted to 50%, 25% or 10% with PBS solution. 0.5 ml of MRSA bacterial solution or Candida bacterial solution was added to the sample solution (4.5 ml), and CFU was calculated using the same method as in the above comparative experiment. As a result, as shown in Table 2-a and Table 2-b, the bactericidal effect (bactericidal rate) against MRSA bacteria (Table 2-a) and Candida bacteria (Table 2-b) of the three types of bamboo vinegars is As the concentration of these test samples increased, the sterilization rate of the 50% electrolytic bamboo acetic acid solution was 83% or more for all bacteria. On the other hand, Candida was more sensitive to three kinds of bamboo vinegar than MRSA.
That is, the resistance of Candida was weaker than that of MRSA. From these results (Tables 1 and 2), the horizontal transmission of drug-resistant bacteria that are resistant to antibiotics in hospitals and nursing homes is a serious issue. Although it exhibits bactericidal properties, it is extremely difficult to avoid the occurrence of side effects and side reactions on living bodies. On the other hand, the antibacterial and bactericidal components derived from natural products are less effective against resistant bacteria, but their extremely low side effects are of great interest both physiologically and bacteriologically.
[0018]
[Table 2]

[0019]
<Method for preparing burned mouse wound site and measurement of burn wound area>
After administering 0.1 ml (24 mg / ml) of thiopental sodium anesthetic (labonal, manufactured by Manyu Pharmaceutical Co., Ltd.) into the abdominal cavity of 7-week-old ICR female mice (average body weight: 32 g; manufactured by CLEA Japan, Inc.) The hair on the back was shaved with an electric clipper, and an electric iron (2 cm × 3 cm, 100 V / 100 W) was applied to the hair removal surface for 3 seconds to prepare a burn wound site (6 cm 2 = 10% / body surface area). This burn wound corresponds to a third-degree burn, which is the limit of survival in humans, or 10% of the body surface area. Immediately after the burn and every 7 days after the burn, cellophane paper was applied to the wound surface of the back of the mouse under anesthesia for 28 days, and the wound surface was traced with a marker on the cellophane paper, and then an image analyzer (Modulator System, MOP-AM03). The wound area was measured at Kontron, Germany).
[0020]
<Example 3>
It was prepared by mixing 1 volume of the electrolytic bamboo acetate solution (made by Tachibana Bamboo Co., Ltd.) of Example 2 with 3 volumes of the share cream (made by IMC Corporation). This 25% electrolytic bamboo acetic acid solution / shear cream combination agent (hereinafter abbreviated as 25% ABV · SC) is referred to as Example 3.
In addition, the electrolytic bamboo acetic acid solution of this ABV / SC compounding agent can be blended even at a concentration range of 5 to 25%. Further, bamboo vinegar, electrolytic bamboo acetate and / or electrolytic bamboo vinegar alkaline solution in the above-mentioned concentration range can be blended in share lotion (manufactured by IMC) in place of shear cream.
[0021]
<Mixed infection test>
Mixed infection experiment system is (1) Formulation cream non-application control group, (2) Shear cream (SC) single application group, (3) Electrolytic bamboo acetate solution (ABV), (4) 25% ABV / SC application group A total of 3 groups (3 for each group) were prepared, and the following experimental mixed infection experiment was conducted.
Immediately after the burn, the necrotic epidermis part of the burn wound surface is peeled off, and 0.1 ml of a mixed bacterial solution in which MRSA bacteria (5 × 107 CFU) and Candida bacteria (5 × 107 CFU) are mixed in equal amounts on the peel surface is smeared and infected with a sterile cotton swab. On the first and third days, 0.5 g each of the above-mentioned shear cream or 25% ABV / SC shear cream is applied and spread with a sterile toothpick. Immediately after smear infection, the infected wound surface was wiped off with a cotton swab dipped in sterile PBS on days 7, 14, 21 and 28, and the swab was transferred to a 0.5 ml PBS test tube and diluted 10-fold in steps. The CFU value of MRSA bacteria in the diluted bacterial solution was calculated with Staphylococcus 110 agar medium, the CFU value of Candida bacteria was calculated with Candida GE agar medium, and the CFU of these infectious bacteria per burn wound area was determined. CFU / cm ² Converted into As a result, the number of viable reduced bacteria (log CFU / cm) from the wound surface of MRSA bacteria and Candida bacteria in the group infected with share cream (SC) alone ² ) In comparison with that of the non-application-infected control group, as shown in the upper and middle stages of Table 3, a decrease trend was observed over 7 to 28 days after infection, but the order value during the experiment period. Was almost the same as those in the control group, and no significant difference was observed between the two groups. On the other hand, the decrease in the number of viable bacteria in the group infected with 25% electrolytic bamboo acetic acid solution (ABV) alone was significant, and was significant after the 14th day of infection. When a significant difference was tested (Student's t-test method), in MRSA infection, there was a significant difference of a risk rate of 5% or less on the 14th day after infection, and a risk rate of 1% or less on the 21st and 28th days (Table: In the case of Candida infection, a significant difference (table: middle) was obtained with a risk rate of 5% or less on the 14th and 21st days of infection and a risk rate of 1% or less on the 28th day. On the other hand, the decrease in these infectious bacteria in the group infected with 25% electrolytic bamboo acetic acid added share cream (ABV / SC) was significantly more significant than that in the group treated with electrolytic bamboo acetic acid alone, MRSA and Candida. In any of the cases, the significant difference on the 14th day after infection was 1% or less of the risk rate, and the significant difference between the two groups after the 21st day was more remarkable (risk rate of 0.1% or less). . On the other hand, when the change of the burn wound surface in the mixed infection of MRSA bacteria and Candida bacteria was examined, as shown in the lower part of the table, the mixed infection bacteria after the 21st day in the 25% ABV / SC application infection group were observed. The reduction in the number of viable reduced bacteria was extremely remarkable (risk rate of 0.1% or less). From these results, it was clarified that the action of eliminating mixed infection bacteria from the burn wound surface is synergistically promoted by adding electrolytic bamboo acetate solution to the shear cream. Moreover, when considering the hardness of the shear cream and the bactericidal action of the electrolytic bamboo acetic acid solution, the ratio of the added amount of the electrolytic bamboo acetic acid solution is 5 to 25% (cream concentration: 95 to 75%). It was thought that it was appropriate to add so that.
Further, even in the case of a BV / SL solution prepared by adding a non-electrolytic bamboo vinegar solution (BV) to a share lotion (SL), the above concentration range (5 to 25%) is considered to be appropriate.
[0022]
[Table 3]

[0023]
<Number of infiltrating cells and classification of infiltrating cells on the wound surface of burns infected with mixed bacteria>
After smear infection of the infection experiment system group 4 (non-application control group and application group 3 group) described in <Mixed infection test> above, 10% fetal bovine serum was added between the dermis layer and the subcutaneous tissue layer on the wound surface on the seventh day After injecting 5 ml of Eagle MEM culture medium (heparin: 4 units / ml) and thoroughly squeezing the back, the injected culture medium was collected with a syringe and 1.0 ml was transferred to a plastic microplate (diameter 30 mm; 6 wells). did. After incubating the microplate at 37 ° C. for 2 hours (5% carbon dioxide incubator), the culture solution was removed. After fixing the microplate with methanol and Giemsa staining, the total number of infiltrating cells in the well and the classification of infiltrating cells were calculated under a microscope (see FIG. 1). As a result, as shown in Table 4: on the left side, the total number of infiltrating cells in the infected group with electrolytic bamboo acetate solution (ABV) alone was observed to increase slightly more than that in the non-coated infection control group, There was no significant difference. On the other hand, the total number of infiltrating cells in the two application groups increased, and the significant difference from the non-application infection control group was less than 5% in the share cream (SC) infection application group and 25% ABV · In the SC application infection group, the risk rate was 1% or less.
On the other hand, when the cell classification of these infiltrating cells was performed, as shown in the table: right side, in the non-coated infection control group, the ratio of lymphoid cells was the highest value (average 89.5%), and the network of macrophages and the like The average number of internal cells was 5.2%, and the epithelial and fibroblast group was only 5.3%. In contrast, macrophage-like cells increased to about 9% in the group infected with electrolytic bamboo acetate solution alone, but there was no significant difference from the cell classification ratio in the non-coated infection control group.
On the other hand, the ratio of macrophage-like cells in the share cream-only-infected group and the 25% ABV / SC-infected group was significantly increased, showing an average of 23.4% and 46.8%, respectively, and the 25% ABV / SC-infected group The rise of the epithelial and fibroblastic cell groups in (13.7% on average) was also significant (see FIG. 1 :).
These results were consistent with the results of the significant effect of promoting the elimination of mixed infections observed after 14th day of the group 2 of application infection in the <mixed infection test> described above. It was strongly suggested that the main component of shear fat has an action of promoting the migration of macrophages, which are the first cells of the immunocompetent cell group, to the damaged site and affected area, and the ability to enhance macrophage function (immunity enhancement ability). Furthermore, since bamboo vinegar contains a large amount of carbohydrates (363 mg / g) and dietary fiber (177 mg / g), these components also activate immunocompetent cells represented by macrophages. The possibility of the occurrence is sufficiently considered from the results in Table 4.
[0024]
[Table 4]

[0025]
<Histopathological examination of subcutaneous tissue of wound surface of mixed bacteria-infected burn>
Next, in order to establish the reasoning described in the above <Number of infiltrating cells in the subcutaneous tissue and classification of infiltrating cells on the wound surface of the mixed bacterial smear infected burn>, the following examination was performed. Among the above four groups, the skin tissue of the mouse wound surface closest to the average wound area value on the 14th day excluding the charged bamboo acetate solution alone application group was excised and treated according to a conventional method. Specifically, tissue sections were prepared after dehydration (alcohol series), fixation (1% glutaraldehyde, 1% paraformaldehyde), and paraffin embedding. Furthermore, after attaching and fixing the treated tissue section on a slide glass, the tissue was stained with an elastica fungison staining solution that can specifically dye connective tissue fibrous protein, and connective tissue (collagen fiber, The degree of regeneration and formation of fibers (elastic fibers, fibers such as collagen and elastin) was observed under a microscope. As shown in FIG. 2, an infectious immune function enhancer, a metabolic function promoter, a biological function deterioration preventive and improving agent When the subcutaneous tissue section immediately after the burn was stained with an elastica fungison stain that allows histological observation of the state of connective tissue, the site of presence, the amount of formation, etc., the layer stained pink was completely The skin tissue itself was in a completely necrotic state, and the influence of the burn corresponding to human third degree burn reached not only the dermis layer but also the subcutaneous tissue layer. On the other hand, in the skin tissue slice image of the non-application control group on day 14 after the burn, infiltration of epithelial (circular) cells and fibroblasts into the dermis layer was remarkable, but it was stained pink. There were very few collagen fibers and elastic fibers (collagen, elastin, etc.), and almost no regeneration of the epidermis layer or dermis layer was observed. On the other hand, when the skin tissue section on the 14th day after the burn in the shear cream (SC) single application group was observed, the regeneration of the epidermis layer was almost completed and it was stained in a bright pink-red color. The site was found from the epidermis layer to the subcutaneous tissue layer, and there were almost no fibroblasts or epithelial cells, but the stained subcutaneous tissue layer was conspicuous and the complete regeneration of the subcutaneous tissue was observed. Was not reached. On the other hand, when the skin tissue section in the 25% ABV / SC application group was observed, the entire skin was stained in deep pink from the dermis layer to the subcutaneous tissue layer, and the denseness of the stained subcutaneous tissue layer was , Significantly more than that of the SC alone application group. From these results, by adding an electrolytic bamboo acetate solution to the share cream, not only the effect of promoting the elimination of opportunistic pathogens on the burn wound surface, but also sugar, dietary fiber and protein (36 mg) which are constituent components of bamboo vinegar solution / G) enhances the functions of epithelial cells and fibroblasts, and at the same time, the main constituents of shear fat also synergistically promote the regeneration of connective tissue, the production of connective tissue components, and thus the metabolic function of living tissues. It was also confirmed from histopathological observation that it would be accelerated.
[0026]
<Reduction / Healing Transition of Mixed Bacterial Smear Infection Burn Wound Area>
The mixed bacterial smear-infected wound surface application experiment system was prepared with 3 groups of mice (10 per group) similar to those described in <Mixed infection test>, immediately after the burn, 4, 14, 21, and 28 after the burn. The wound area on the day was measured by the method of Example 2. As a result, as shown in Table 5, when the wound area immediately after the burn was measured, it was 6.53 ± 0.41 cm 2, and the surface area of the electric iron (6.0 cm ² ) Was created. When an experiment was performed using such a burn wound surface, the wound areas on the 7, 14, 21 and 28th days after the burn in the group infected with electrolytic bamboo acetate (ABV) alone averaged 6.06 on average, 4.9, 2.16 and 1.01 cm 2, which gradually decreased with the passage of days and showed lower than the mean wound area at 7-28 days in the non-application infected control group, but the significant difference was 28 days It was only observed in the eyes (risk rate 5% or less). On the other hand, in the share cream (SC) single application infection group, a significant difference (risk rate of 5% or less, area: 5.46 ± 0.49 cm) on the seventh day after the burn. ² ) Was observed, and after 14th day, a further significant difference (risk rate of 1% or less) was shown. On the other hand, the reduction (healing) of the wound area in the 25% ABV / SC application-infected group was significantly promoted compared with the previous two groups, and the wound with extremely high significance (risk rate of 1% or less) on the seventh day after the burn. Area reduction (4.81 ± 0.29cm ² ) Was observed, and the risk rate after the 14th day was 0.1% (see FIG. 3).
Summarizing these results, by adding an electrolytic bamboo acetate solution to the share cream, the healing of the original dermatitis and skin diseases of the share cream can be achieved by eliminating and sterilizing the mixed infection of the electrolytic bamboo acetate solution. It was fully suggested that the effect may have been synergistically and efficiently exerted on wound healing of infected burns.
[0027]
[Table 5]

[0028]
<Toxicity test>
Non-electrolytic bamboo vinegar solution, electrolytic bamboo acetic acid solution, shear cream, shear lotion, and these vinegar-added shear cream and / or share lotion healthy test (40 people), ie, primary skin irritation test, light sensitivity A cropping property (allergy) test and a phototoxicity test were conducted. On the other hand, no toxic reaction was observed in the toxicity test by continuous oral administration (once / day) of non-electrolytic bamboo vinegar solution and electrolytic bamboo acetic acid solution using ICR mice (Chart: omitted).
[0029]
<Example 4>
Warm / burn wound poultice and / or ointment
10 g of the share cream (manufactured by IMC Co., Ltd.) is mixed with the electrolytic bamboo acetic acid solution in Table 6 below, a rice-derived sugar ceramide and multivitamins (A, E, C), and 90 g of the share cream. In addition, a poultice for scalding / burning wound surface infection bacteria removal and wound surface healing promotion with a total content of shear cream of 100 g was produced.
Similarly, 10 g of the share cream (manufactured by IC Co., Ltd.) is mixed with the electrolytic bamboo acetic acid solution of the following Table 6 [ointment], rice-derived sugar ceramide and multivitamins (A, E, C). In addition, 100 g of white petrolatum was added to prepare an ointment for eliminating wrinkle / burn wound surface infection bacteria and promoting wound surface healing.
[0030]
[Table 6]
[Poultice]
Share cream 100g
Electrolytic bamboo acetate solution 25ml
Rice-derived sugar ceramide 100mg
Multivitamin (A, E, C) 500mg
White petrolatum 0g
[Ointment]
10g share cream
Electrolytic bamboo acetate solution 25ml
Rice-derived sugar ceramide 100mg
Multivitamin (A, E, C) 500mg
100g white petrolatum
[0031]
<Example 5>
The ingredients shown in Table 7 below were mixed with 10 ml of a share lotion (manufactured by IMC), and 90 ml of the share lotion was further added to prepare a lotion for use in preventing stains, wrinkle formation prevention / erasure promoters, and cosmetics.
[0032]
[Table 7]
[Medical and cosmetic lotion]
Share lotion 100ml
Non-electrolytic bamboo vinegar solution 10ml
Rice sugar ceramide 50mg
Multivitamin (A, E, C) 100mg
[0033]
<Example 6>
Add the appropriate amount of glucose solution and fragrance to non-electrolytic bamboo vinegar solution, sparingly soluble oligosaccharides and multivitamins, and use 100 ml of sterilized purified water to promote metabolism function and prevent deterioration of various biological functions (functions) Food).
This functional food (the amount of components contained in 100 ml) is shown below.
[0034]
[Table 8]
[Functional food]
Non-electrolytic bamboo vinegar solution 10ml
Slightly soluble oligosaccharide 2g
Multivitamin 300mg
[0035]
【The invention's effect】
The invention of the present application can provide a drug comprising a bamboo acetic acid solution and a shear fat derived from a sheared tree fruit and / or an extract / purified fraction composition thereof as essential components. Antibacterial and / or bactericidal effects against mixed bacterial infections for The invention of claim 2 can provide a skin tissue metabolic function promoter, and the invention of claim 3 can provide a damaged skin tissue regeneration function promoter. The invention of claim 4 is , Infectious immunity function-enhancing agent can be provided.
[Brief description of the drawings]
FIG. 1 is an image of infiltrating cells in a burn wound surface subcutaneous tissue on the seventh day after burn / mixed infection according to an example of the present application.
FIG. 2 is a subcutaneous tissue section image of a burn wound surface 14 days after burn / mixed infection according to an example of the present application.
FIG. 3 is a reduction and treatment image of a burn wound surface on the 21st day after burn / mixed infection according to an example of the present application.

Claims

Bamboo vinegar and / or bamboo acetic acid obtained from the anode after electrolysis, bamboo shear vinegar-derived share fat and / or extract and purified fraction composition thereof as essential components, An antibacterial and / or bactericidal agent for opportunistic mixed infections, characterized in that the concentration of the sex fluid is 5 to 25%.

The bamboo acetic acid and / or bamboo acetic acid obtained from the anode after the electrolysis treatment and the share-fruit-derived shear fat and / or its extracted and purified fraction composition are essential ingredients, and this bamboo acetic acid A skin tissue metabolic function promoter, characterized in that the concentration of the sex fluid is 5 to 25%.

Bamboo vinegar and / or bamboo acetic acid obtained from the anode after the electrolysis treatment, shear tree fruit-derived shear fat and / or its extracted and purified fraction composition as essential components, and this bamboo acetic acid An agent for promoting regeneration of damaged skin tissue, which is formulated so that the concentration of the liquid is 5 to 25%.

Bamboo vinegar and / or bamboo acetic acid obtained from the anode after the electrolysis treatment, shear tree fruit-derived shear fat and / or its extracted and purified fraction composition as essential components, and this bamboo acetic acid An infectious immunity function-enhancing agent, which is formulated so that the concentration of the liquid is 5 to 25%.