JP5449780B2 - 酸化還元電位水溶液を用いた炎症及び過敏症の治療又は予防方法 - Google Patents
酸化還元電位水溶液を用いた炎症及び過敏症の治療又は予防方法 Download PDFInfo
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Description
本特許出願は、2006年1月20日に出願された米国仮特許出願第60/760,635号;2006年1月20日に出願された同第60/760,567号;2006年1月20日に出願された同第60/760,645号;及び2006年1月20日に出願された同第60/760,557号の利益を主張しており;これら全ては参照によって全体として本明細書に組み込まれる。
本発明は、治療有効量の酸化還元電位(ORP)水溶液を患者に投与することによる、患者の炎症の予防又は治療の方法を提供し、ここで該溶液は少なくとも24時間安定である。本発明の方法は、様々な原因因子(例、アレルギー反応、自己免疫反応、感染、1種以上の炎症誘発物質との接触、及びそのような原因因子の組み合わせ)に起因する炎症の治療において使用され得る。
本発明は、患者における炎症を予防又は治療する方法を提供し、当該方法は該患者に治療有効量の酸化還元電位(ORP)水溶液(超酸化水(SOW)としても知られる)を投与することを含み、該溶液は少なくとも約24時間安定である。本発明の方法は、過敏症(例えばアレルギーにおける過敏症など)を含む急性炎症及び慢性炎症の治療又は予防(例、発症の阻害、進行の阻害、可能性の減少)のために使用することが出来る。本発明の方法に従って治療可能又は予防可能な炎症及び過敏症としては、例えば有害刺激との接触、負傷、感染、自己免疫反応、過敏症及びアレルギー反応(細胞性ヒスタミン及び炎症促進性サイトカインの放出に関連するアレルギー反応を含む)に起因する炎症を挙げることが出来る。
小規模用途のために、ORP水溶液は、スタンドパイプ及びポンプを含むスプレーボトルを通じて分配され得る。或いは、ORP水溶液は、エアロゾル容器中に詰められ得る。エアロゾル容器は、分配される製品、推進剤、容器及びバルブを含み得る。バルブは、アクチュエータ及び浸漬チューブの両方を含み得る。容器の内容物は、アクチュエータを下に押すことによって分配され得る。エアロゾル容器の種々の成分は、ORP水溶液と適合性があるべきである。好適な推進剤としては、液化ハロカーボン、炭化水素、又はハロカーボン−炭化水素混合、或いは二酸化炭素、窒素又は亜酸化窒素などの圧縮気体を挙げることができる。エアロゾルシステムは、好ましくは、寸法が約0.15μmから約5μmに及ぶ液滴を与える。
これらの実施例は、本発明に従って用いられるORP水溶液の独自の特徴を示している。実施例1−3のORP水溶液のサンプルを、本明細書に記載の方法に基づいて分析し、各サンプルに存在するイオン種及び他の化学種の物理的特性及びレベルを決定した。二酸化塩素、オゾン及び過酸化水素について得られた結果は、そのような種を測定するために用いられる標準試験に基づいているが、これらの結果は、肯定的な試験結果を生じることもあり得る様々な種を示すものであり得る。さらに、二酸化塩素、オゾン及び過酸化水素が次亜塩素酸塩(hypocholrite)と反応し、これらの消費及び他の化合物(例、HCl及びO2)の生成をもたらすことが報告されている。ORP水溶液の各サンプルについて、pH、酸化還元電位(ORP)及び存在するイオン種を表1に示す。
これらの実施例は、本発明に従う、ORP水溶液への種々の量での漂白剤の添加について示している。特に、これらの実施例は、組成物の抗菌活性及び織物を漂白する能力を示している。
本研究の目的は、ウサギの鼻腔に滴として投与した場合の、被験(test)である例示的なORP水溶液であるMicrocynの安全性を評価することであった。33匹のウサギを、群I及び群IIの2つの群に無作為に割り当てた。群I(18匹)を対照群とし、群II(15匹)には被験物質を投与した。−1日目又は0日目に、体重を記録し、選択したパラメーターの分析のために血液サンプルを集めた。0日目に、群Iの動物に500μLの無菌食塩水を投与し、群nのアニュアル(annual)に500μLの被験物質(50%濃度)を投与した。対照及び被験物質は共に、右鼻孔に滴として1日2回投与した。1日目−6日目に同じ方法で動物に投与した。鼻に特に注意を払って、薬理的及び/又は毒性効果の兆候について、動物を毎日観察した。研究が終わるまで体重を毎週記録した。7日目に各群の3分の1の動物を、採血、屠殺及び剖検のために選抜した。残りの動物には14日目まで投与を継続し、14日目に各群の半分の動物を、採血、屠殺及び剖検のために選抜した。21日目(7日の回復期間後)に、残りの動物を採血し、屠殺し、剖検した。両鼻孔由来の鼻粘膜サンプルを、病理組織学的分析のために各動物から採取した。
本実施例は、例示的なORP水溶液の使用では、毒性がないことを示している。
本実施例は、咽頭炎の治療に対する例示的なORP水溶液の有効性を決定するために使用できる臨床研究を示している。
本実施例は、アデノウイルス血清型5に対する例示的なORP水溶液の殺ウイルス(viricidal)活性を示している。本実施例では、E1a、部分的にE1−b、及び部分的にE3欠損であるヒトアデノウイルス5型に基づいたアデノウイルス(Ad)ベクターを用いた。pCMVの転写調節下の緑色蛍光タンパク質(GFP)レポーター遺伝子を含むシャトルプラスミドを調製した(pAd−Track)。このpShuttleプラスミドとAdEasy1プラスミドとの相同的組み換えをエレクトロコンピテント細菌内で行った。挿入を有するクローンを、制限エンドヌクレアーゼ消化により試験した。確認されたら、大規模な増幅のために、超らせんプラスミドDMAをDH10B細胞に形質転換した。続いて、293細胞(ATCC 1573)を無血清培地(OptiMEM−GIBCO)で培養し、Padで消化した組み換えプラスミドでトランスフェクトした。細胞変性効果について感染細胞をモニターし、それらを収集し、凍結解凍を3サイクルして溶解した。得られたウイルス(AdGFP)を製造者の使用説明書に従ってAdenoPureカラム(BD Clontech)で精製した。ウイルスをOD260/280で定量した。最終収量は1.52X1011pfu/mLであった。
本実施例は、無生物の環境表面の殺菌に関する米国環境保護局のプロトコルを用いて、HIVに対する例示的なORP水溶液の殺ウイルスの有効性を示している。
本実施例は、過酸化水素(HP)に対する例示的なORP水溶液の、ヒト2倍体線維芽細胞(HDF)の生存能力に対する影響を示している。この潜在的な毒性を試験するために、HDFをインビトロでORP水溶液及び過酸化水素(HP)に曝露した。HPは真核細胞に対して毒性であることが知られており、アポトーシス及びネクローシスを増加させ、細胞の生存能力を減少させる。本実施例において、細胞の生存能力、アポトーシス及びネクローシスを、純粋なORP水溶液及び880mMのHP(HPの消毒用途で採用される濃度)に5分及び30分間曝露したHDFにおいて測定した。
本実施例は、HDFにおける酸化的DNA損傷及びDNA付加体8−ヒドロキシ−2’−デオキシグアノシン(8−OHdG)の形成への、過酸化水素(HP)と比較した例示的なORP水溶液の影響を示している。細胞内での8−OHdG付加体の生成は、DNAの特定の残基における酸化的損傷のマーカーであることが知られている。さらに、この付加体の高い細胞内レベルは、突然変異生成、発癌及び細胞の老化と相関がある。
本実施例は、HPと対比した、低濃度の例示的なORP水溶液への慢性曝露の、HDFへの影響を示している。慢性の酸化ストレスは、細胞の早期老化を引き起こすことが知られている。長期の酸化ストレスを模倣するために、20集団の倍増の間、初代HDF培養物を低濃度のORP水溶液(10%)又はHP(5μM)に慢性的に曝露した。SA−β−ガラクトシダーゼ酵素の発現及び活性は、以前からインビボ及びインビトロでの老化プロセスと関連付けられてきた。本実施例においては、SA−β−ガラクトシダーゼ酵素の発現を、ORP水溶液又はHPに対するHDFの1ヵ月の継続的な曝露後に解析した。結果を図6に示す。酵素SA−β−ガラクトシダーゼの発現を、20の顕微鏡視野における青色の細胞の数をカウントすることにより解析した。図6は、SA−β−ガラクトシダーゼを過剰発現している細胞の数により示されるように(n=3)、HP処理のみが細胞の老化を加速させたことを示している。低用量のHPでの慢性的な処理は、86%の細胞においてSA−β−Galの発現を増加させたが、ORP水溶液での処理はこのタンパク質の過剰発現を引き起こさなかった。ORP水溶液は細胞の早期老化を引き起こすものではないということが本実施例より結論付けられ得る。
本実施例は、腹膜炎患者における、腹膜の細菌負荷の減少及び入院の長さの減少に対する例示的なORP水溶液の効果を示している。2004年6月から2005年1月までにメキシコシティーのRuben Lenero病院に入院し、且つ、急性の汎発性の続発性腹膜炎と診断された全ての患者をORP水溶液治療群に含んだ。続発性腹膜炎は、腹膜腔の汚染に繋がる胃腸又は尿生殖路の完全性の喪失の結果と定義した。同施設において2003年から2004年までの間に同様の腹膜感染症を示した対照症例(paired -case)の遡及的解析を対照群について行った。20人の継続患者を、予めORP水溶液治療群(即ち試験群)に含めた。
本実施例は、例示的なORP水溶液(Mycrocyn)の、肥満細胞の脱顆粒の阻害における有効性を示している。肥満細胞は、I型過敏性疾患において役割を果たす主要なものとして認識されてきた。アトピー性皮膚炎、アレルギー性鼻炎及びアトピー性喘息において観察される複数の臨床症状は、異なる罹患組織にある肥満細胞のIgE抗原刺激によって引き起こされる。アトピー性喘息の発症機序について現在認められている見解は、アレルゲンが、IgE産生肺肥満細胞(MC)を誘発することによってプロセスを開始させて、いわゆる即時相反応においてヒスタミン、ロイコトリエン、プロスタグランジン、キニン、血小板活性化因子(PAF)などのメディエーターを放出させるということである。続いて、これらのメディエーターは、気管支収縮を引き起こし、血管透過性及び粘液産生を亢進する。このモデルによれば、肥満細胞活性化に続いて、それらの細胞は腫瘍壊死因子アルファ(TNF−α)、IL−4、IL−5及びIL−6を含む様々な炎症促進性サイトカインを遅延相において分泌し、それらが好酸球、抗塩基球、Tリンパ球、血小板及び単核食細胞などの他の炎症性細胞の局所的動員及び活性化に関与する。次に、これらの動員された細胞が、その後自律性となる可能性があり、また、喘息の症状を悪化させる可能性がある炎症反応の進行に寄与する。この遅延相反応は、周囲組織における可塑的な変化を誘導し得る長期の炎症プロセスを構成する(Kumar et al., pp. 193-268参照)。
本実施例は、例示的なORP水溶液の、カルシウムイオノフォアによる肥満細胞の活性化に対する阻害活性を示している。
本実施例は、例示的なORP水溶液の、肥満細胞のサイトカイン遺伝子の転写の活性化に対する影響を示している。
本実施例は、例示的なORP水溶液の、肥満細胞のTNF−α分泌に対する阻害活性を示している。
本実施例は、例示的なORP水溶液の、肥満細胞のMIP 1−α分泌への阻害活性を示している。
この実施例は、例示的なORP水溶液を使用した毒性調査の結果を示している。
この実施例は、例示的なORP水溶液の潜在的な細胞遺伝毒性を決定するために行った調査を表している。
この調査は、例示的なORP水溶液Dermacynには毒性がないことを示している。
この調査は16匹のラットで行い、例示的なORP水溶液Dermacynの局所耐容性、及び全層皮膚創傷治癒のモデルにおける創傷床の組織病理への影響を評価した。創傷を対象ラットの両側に作った。治癒過程の間、皮膚切片を左側又は右側のいずれかに置いた(例えば、それぞれDermacyn処理及び食塩水処理)。
本実施例は、例示的な酸化還元電位水Microcynの有効な抗菌性溶液としての本発明に従った使用を示している。
本実施例は、HIBICLENS(登録商標)グルコン酸クロルヘキシジン溶液4.0%(w/v)及び0.9%塩化ナトリウム洗浄液(USP)と対比した、本発明に従って用いた例示的な酸化還元電位水Microcynの抗菌活性の比較を示している。
本実施例は、ペニシリン耐性ストレプトコッカス・ニューモニエ(ATCC 51915)に対するORP水溶液の有効性を示している。
本実施例の目的は、細菌懸濁液アッセイを使用して、バシトラシンと対比した、例示的なORP水溶液(Dermacyn)の微生物活性を決定することである。
本実施例は、バイオフィルム中の細菌に対する2つの例示的なORP水溶液(M1及びM2)の有効性を示している。
Claims (6)
- アトピー性皮膚炎の治療又は予防のための薬剤を製造するための、酸化還元電位水溶液の使用であって、該水溶液は少なくとも2ヶ月安定であり、該水溶液は6.4から7.8のpHを有し、該水溶液はカソード水とアノード水との混合物を含み、該水溶液は10ppm〜400ppmの遊離塩素種を含み、かつ、該水溶液は、次亜塩素酸、次亜塩素酸イオン、次亜塩素酸ナトリウム、亜塩素酸イオン、塩化物イオン、溶解塩素ガス、二酸化塩素及びこれらの混合物からなる群から選択される少なくとも1種の遊離塩素種を含み、該酸化還元電位水溶液が肥満細胞の脱顆粒を阻害し、それによってアトピー性皮膚炎を治療する、使用。
- 酸化還元電位水溶液のpHが7.4から7.6である、請求項1記載の使用。
- 酸化還元電位水溶液が、10体積%から50体積%のカソード水及び50体積%から90体積%のアノード水を含む、請求項1記載の使用。
- 酸化還元電位水溶液が、15ppmから35ppmの次亜塩素酸、25ppmから50ppmの次亜塩素酸ナトリウムを含む、請求項1記載の使用。
- 酸化還元電位水溶液が、−400mVと+1300mVとの間の電位を有する、請求項1記載の使用。
- 薬剤が、抗生物質、抗ウイルス剤及び抗炎症剤、並びにそれらの組み合わせからなる群から選択される少なくとも1つの治療剤をさらに含む、請求項1記載の使用。
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