JP2021519812A - 抗菌性光増感剤の組成と方法 - Google Patents
抗菌性光増感剤の組成と方法 Download PDFInfo
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Abstract
Description
光線力学療法(PDT)は、活性酸素種(ROS)の形成を誘発します。治療の抗菌光線力学的効果は、光増感剤の物理的および化学的パラメーター、例えば、吸収ピーク(kmax)、吸収強度(emax)、および一重項酸素の量子収率に依存します。タイプI光線力学療法では、活性化された光増感剤が電子を伝達してフリーラジカルを形成し、これが活性酸素種(ROS)を形成します。ラジカルは酸素と相互作用してスーパーオキシドアニオンラジカル(O_−2)を生成します。これは細胞構造に酸化的損傷を引き起こす可能性があるため、細胞毒性があります。対照的に、タイプII光線力学療法では、活性化光増感剤はそのエネルギーを直接酸素に伝達して、活性酸素種(ROS)である一重項酸素(1O2)を形成します。生成された活性酸素種は、さまざまな細胞基質(すなわち、細胞膜、DNA、ミトコンドリア、リソソーム)を酸化し、それによって細胞毒性を示します。一重項酸素の寿命は非常に短く(〜3μs)、細胞毒性効果を生み出すには細胞のすぐ近くで生成する必要があります。メチレンブルーの場合、II型反応が起こると仮定されています。
抗菌剤は、微生物を殺す、阻止する、または増殖を停止させる化学種、組成物、または化合物である。抗菌剤は、細菌(抗菌剤)や真菌(抗真菌剤)を含むがこれらに限定されない、さまざまな感染を引き起こす微生物に対して活性を示す可能性があります。抗真菌剤、または殺菌剤は、真菌、真菌構造、または真菌胞子を殺すか、または増殖を阻害するために使用される殺生物性化合物、組成物、化合物のグループ、または生物である。海洋無脊椎動物、植物、および他のさまざまな天然資源からの化合物および代謝産物を含む、多種多様な天然抗真菌剤が知られている。天然の抗真菌性化合物は、シナモジアルステロール、フラノン、キニーネ、レゾルシノール、テルペノイドなど、さまざまな構造クラスに分類されます。天然源に由来する化合物の抗真菌活性は、通常、源のいくつかの有効成分内で達成され、通常、合成化合物と比較して副作用が少なく、より安全です。
光線力学療法のために選択された光源は、最大の効果を有するために、光増感剤の吸収または励起に対応する波長で放出されるべきである。光源は、光線療法治療専用の装置に組み込むことができます。特に、光源は体にエネルギーを与え、光増感剤の存在下で微生物の分解を引き起こします。光増感剤の励起波長は、使用される特定の光増感化合物および光強度に応じて変化し得る。光増感剤の吸収の波長帯もまた、皮膚の光毒性を回避するために、好ましくは600〜800nmの間であるべきである。この治療域は、典型的な日光(波長400〜600 nm)への曝露中の吸収を最小限に抑え、800nmを超える波長で増加する水分子による光吸収を最小限に抑えます。(Plaetzer et al.、LasersMed。Sci.24、pp.259−268、2009;Sekkat et al.、Molecules、17、pp.98−144、2012)光源はオプションでLED(発光ダイオード)です。光源。コヒーレント(レーザー)および非コヒーレント(ダイオード発光−LEDおよびランプ)光源の両方をPDTに使用できます(NymanおよびHynninen、J.Photochem。Photobiol。BBiol.73、pp.1−28、2004)。一例では、クロロフィルaは600〜700nmの範囲の吸収帯を有し、クロロフィルbは600〜675nmの範囲の吸収帯を有する。光源はまた、真菌の成長または死の遅延を引き起こし、治療結果を改善するために、光増感剤の用量およびすべての要因の最適な組み合わせに基づいてエネルギー密度を変化させることができる。レーザーは、光ファイバに焦点を合わせることができる高度な単色性の光を供給することができます。ただし、PDTでレーザーを使用する場合の欠点のいくつかは、コストが高く、輸送が難しいことです。LEDはより安価で、持ち運びが容易であり、より長い波長の光増感剤の発見により、PDTの実験的および臨床的応用でますます使用されています(Hamblin et al.、CancerRes.56、pp.5205−5210、1996;Nyman and Hynninen、J.Photochem.Photobiol.BBiol.73、pp.1−28、2004)。白色または蛍光灯の場合、宿主組織を加熱するリスクを最小限に抑えるために、赤外線だけでなく突然変異誘発を避けるために紫外線放射を最小限に抑えることが重要です(Donnelly et al.、Microbiol.Res.163、pp.1−12、2008)。さらに、赤色光は約3〜4nmを透過するのに対し、青色光は約1.5nmを透過することがわかっています(Donnelly et al.、Microbiol.Res.163、pp.1−12、2008;Garland et al.、FutureMed.Chem.1、pp.667−691、2009)。メチレンブルーの活性化には、単色の赤色光が好ましい。他の光増感剤の場合、治療光の単色波長の選択は、最大吸収ピークまたは光増感剤に依存します。
本明細書に記載の組成物の貯蔵寿命は、患者に様々な治療オプションを提供し、これは、局所微生物感染症の治療に有用である。爪真菌症の場合、治療の必要性は一般に数週間から数ヶ月であり、特別な貯蔵を必要としない長い貯蔵寿命は貯蔵された溶液の改善された効力を提供するので、安定した組成物は特に有用である。ビヒクル組成物は、水性成分、有機溶媒ベースの成分、または水性成分と有機または溶媒ベースの成分との組み合わせを含むことができる。多相組成物は、水相と有機相の両方を含み得る。有機または溶媒成分は極性または非極性であり得る、そして組成物は1つ以上の乳化剤または追加の溶媒または成分をさらに含み得る。一実施形態では、組成物は、1〜50v/v%のグリセリンを含む。イソプロパノール、エタノール、またはそれらの混合物(5〜90v/v%)、および水(10〜90v/v%)。好ましくは、ビヒクルは、10〜50v/v%のグリセリン、5〜35v/v%のイソプロパノールまたはエタノールまたはそれらの混合物、および25〜75v/v%の緩衝液を含む。別の実施形態では、組成物中のエタノールおよび/またはイソプロパノールの量は、10%〜90%の範囲で変化することができ、エタノールは、別の適切なアルコールまたは薬学的に許容される溶媒で置き換えることができる。光増感剤の量は、組成物中、0.01%から10%まで変化し得る。1つの好ましい光増感剤はメチレンブルーである。イヌラビスコサ抽出物の量は、組成において、例えば、0.1%〜10%の間で変化し得る。水の量は10%から90%まで変化し、緩衝液の形をとることができます。一実施形態では、緩衝液はPBS緩衝液である。組成物中のグリセリンの量は、1%〜50%の間で変動し得る。乳化剤はまた、例えば、0%〜5%の量で組成物に添加され得る。クリニックでのテストによると、これらのビヒクルは、組成物の粘度が高く、表面が低いため、水や緩衝液と比較して、爪への浸透が良く、爪への広がりが良く、一般に扱いやすいことがわかっています。テンション。
爪真菌症の患者を治療するために、最初に、薬学的に許容されるビヒクルに処方された、天然由来の抗菌剤、光増感剤を含む組成物で爪を治療する。次に、組成物は、それが吸収されるか、または感染部位に移動することができるように、十分な浸透時間を与えられる。組成物が爪を感染部位に浸透すると、その部位は、光増感剤によって吸収された波長の光源で照射される。光増感剤を光励起すると、活性酸素種が生成され、感染部位の微生物が破壊されます。光は釘を透過することができるので、照射ステップは、場所に直接(釘を取り除いて)、または光増感剤によって吸収された波長の光源を釘を通して間接的に照射して、微生物を破壊することによって達成することができる。感染の場所。この領域はさらに、皮膚治療、組織修復、および/または創傷治癒に有効であることが知られている第2の波長の光で治療することができる。光増感剤の吸収の波長帯において第1の波長で光を放出するための第1の光源と、組織修復、創傷治癒を促進するための第2の波長で光を放出するための第2の光源を提供する光線力学療法のための装置も使用できる。またはそれらの組み合わせ。第2の波長は、例えば、近赤外線、赤、青、および緑の範囲であり得る。
着色光増感剤による爪の治療に続いて、脱色溶液または製剤を使用して、爪から色を除去することができる。たとえば、メチレンブルー+の脱色または色の中和は、CSN−(部分)、H2O2、紫外線、アスコルビン酸、Ce(IV)などの強力な酸化剤、塩基性媒体中の−OHまたはH2O2、硫化物(Na2S)、および他の還元糖および還元剤、単独または組み合わせて使用。例えば、メチレンブルーの青色の中和または脱色は、アルカリ性の水溶液およびグルコースまたはデキストロースなどの還元剤を使用して爪上で起こり得る。処理組成物の一部として皮膚に適用される着色光増感剤を脱色するために使用できる追加の還元剤には、過酸化水素(H2O2)、ギ酸(HCOOH)、アスコルビン酸(C6H8O6)、一酸化炭素(CO)またはCO2、およびその他が含まれる還元剤。一例では、中和溶液は、グルコース−3%およびKOH−2.7%から調製され、pHは約8〜10の範囲である。
担体は、3つの成分、グリセロール、エタノール、および溶媒としての水からなる。グリセロールの場合、6つの体積パーセントが選択されます:60%、30%、15%、7.5%、3.75%、および0。エタノールの場合、6つの体積パーセントが選択されます:40%、20%、10%、5%、2.5%、および0。水については、その体積パーセントにグリセロールとエタノールの体積パーセントを加えたものが、すべての組み合わせで100%でした。この実験では、T.rubrumと2XRPMI培地の体積比は1:19でした。化合物を96ウェルマイクロタイタープレートのウェルに添加して、成長を決定するためにOD値を測定した。結果を表1に示す。グリセリンは、ネイル用途のための組成物の最良の表面張力および粘度を提供するという観点から、30%であることが好ましい。以下の結果と上記の要件に基づいて、抗菌効果の付加価値を得るために、最適化されたエタノール濃度は10%〜40%の範囲にあります。グリセリンは真菌の成長に有益な効果をもたらす可能性があり、プロピレングリコールで置き換えることができます。
滅菌した爪片を新鮮な紅色白癬菌の芝生に置き、背側を上にして少なくとも10日間感染させた。ネイルピースは、0.03125%〜0.25%の範囲の濃度のキャリアでメチレンブルー(MB)で処理されました。対照はメチレンブルーを含まない担体溶液であった。別の実験では、MBのみと光のみでは、真菌の増殖を殺したり阻害したりしませんでした。爪は30分間赤色光(640nm、9W−3つのLEDライト、それぞれ3W)にさらされました。対照と比較した治療後のTrichophytonrubrumの生存可能な真菌コロニーの量を顕微鏡を使用して観察し、一度視覚を肉眼で監視し続けた。結果を表2に示します。ここで、「G」は成長を示し、「N」は成長がないことを示します。示されているように、3日後、対照は真菌の増殖を示した。5日後、0.0625%および0.03125%の処理で菌糸体の成長が見られました。治療の6日後、菌糸体は0.125%の治療で発見されました。治療の12日後、0.25%MBは成長を示しました。再成長の長期阻害を得るために、二次治療または代替治療の必要性があることを意味します。
爪真菌症の長期治療溶液として、イヌラビスコサの使用を調査した。波長640nmでの光線力学療法による一次治療後のInulaViscosaによる爪真菌症の二次治療は、真菌の再増殖の長期阻害を示すことが示された。
追加の光増感剤がない場合、イヌラビスコサが光増感剤として機能し、光線療法効果を有することができることが見出された。理論に拘束されることなく、この効果はおそらくクロロフィルのオグルマ安定化、および/または光線療法の存在下で光増感剤として作用することができるオグルマビスコサの葉に見られる独特のクロロフィルによるものであると仮定されています。
以下の成分を含む2つの別個の相で、組成物を調製した:メチレンブルー(0.25%)、イヌラビスコサ抽出物(1%)、水(48.25%)、エタノール(20%)、グリセリン(30%)。および非イオン性乳化剤(Lipowax D、0.5%)。水相は、追加の光増感剤としてのメチレンブルーを脱イオン水とエタノールの混合物に混合することによって調製された。参考までに、追加の光増感剤なしで同様の組成物を調製することができる。疎水性相は、Lipowax D、Inula viscosa、グリセリンなどの非イオン性乳化剤を混合して調製しました。次に、疎水性相を水相に加え、安定したエマルジョンが形成されるまで激しく混合した。エマルジョンは使用するまで室温で保存した。
爪真菌症は、以下のプロトコルを使用して多くの患者で治療されてきた。治療に関するデータが収集され、診療所への訪問回数が最小限で済み、治療が長引くことによるコンプライアンスの問題がないため、患者への影響は肯定的で効果的でした。このプロトコルおよび記載された組成物および方法は、足指の爪真菌を迅速、効果的、および安全に治療することができる。本明細書に記載の本発明の組成物および方法は、これらおよび他の必要性を満たす。爪真菌症の治療のための治療のための例示的なプロトコルは以下の通りである:
1.爪の背側をそっとやすりで磨きます
2.本発明の抗真菌性組成物を爪に適用し、それが浸透するのを10分間待つ。
3.微生物を破壊するために、抗真菌性組成物の光増感剤によって吸収された波長の光源を爪に照射する。
Claims (26)
- 抗菌性感光性組成物であって、
イヌラビスコサ抽出物と、
薬学的に許容されるビークルと、
を含む、抗菌性感光性組成物。 - 請求項1記載の組成物であって、追加の光増感剤をさらに含む、組成物。
- 請求項2記載の組成物において、前記追加の光増感剤は、ヒペリシン、クロロフィル、クルクミン、メチレンブルー、およびそれらの組み合わせのうちの1つである、組成物。
- 請求項2記載の組成物において、前記追加の光増感剤は、ポルフィリン、フタロシアニン、フェノチアジニウム、ベンゾポルフィリン、ヘマトポルフィリン、ピロール、テトラピロール化合物、ピロリック大環状化合物、ポルフィマー、および5−アミノレブリン酸のうちの1つである、組成物。
- 請求項1〜4のいずれか一項記載の組成物において、前記組成物は、シンナモジアルステロール、フラノン、キニーネ、レゾルシノール、またはテルペノイドからなる群から選択される追加の天然由来抗菌剤をさらに含む、組成物。
- 請求項1〜5のいずれか一項記載の組成物において、前記イヌラビスコサ抽出物は、0.1%〜10%v/vで組成物中に存在する、組成物。
- 請求項1〜6のいずれか一項記載の組成物であって、浸透促進剤をさらに含む、組成物。
- 請求項1〜7のいずれか一項記載の組成物において、前記イヌラビスコサ抽出物は、組成物中の抗菌剤および光増感剤の両方として作用する、組成物。
- 請求項1〜8のいずれか一項記載の組成物において、防腐剤、キレート剤、またはその両方をさらに含む、組成物。
- 請求項1〜9のいずれか一項記載の組成物において、前記薬学的に許容されるビヒクルは、
1−50v/v%のグリセリンと、
イソプロパノール、エタノール、または5〜90v/v%のそれらの混合物と、および
10−90v/v%の水と、
を含む、組成物。 - 請求項1〜10のいずれか一項記載の組成物であって、合成抗真菌剤をさらに含む、組成物。
- 請求項1〜11のいずれか一項記載の組成物であって、抗炎症化合物をさらに含む、組成物。
- 微生物感染症を治療するための光線力学的方法であって、
光増感剤組成物を微生物感染部位に適用する工程であって、前記光増感剤組成物は、イヌラビスコサ抽出物および薬学的に許容されるビヒクルを含む、適用する工程と、
前記微生物感染部位を、前記イヌラビスコサ抽出物の吸収波長帯の波長を有する光に曝露する工程と、
を含む、光線力学的方法。 - 微生物感染症を治療するための光線力学的方法であって、
光増感剤組成物を微生物感染部位に適用する工程であって、前記光増感剤組成物は、天然由来の抗真菌剤、合成光増感剤、および薬学的に許容されるビヒクルを含む、適用する工程と、光線力学的方法。
前記光増感剤の吸収波長帯の波長を有する光に前記微生物感染部位を曝露する工程と、
を含む、 - 請求項13または14記載の方法において、前記微生物感染は、真菌、細菌、ウイルス、いぼ、および皮膚糸状菌のうちの少なくとも1つによって引き起こされる、方法。
- 請求項13〜15のいずれか一項に記載の方法において、前記微生物感染は、爪真菌症である、方法。
- 請求項13〜16のいずれか一項記載の方法において、前記イヌラビスコサ抽出物の吸収の波長帯は、600〜800nmであり、および前記光源は、600〜800nmの波長で発光する、方法。
- 請求項13〜17のいずれか一項記載の方法であって、前記光増感剤組成物を還元して光増感剤を変色させることをさらに含む、方法。
- 請求項13〜18のいずれか一項記載の方法において、前記光は、LED、レーザー、およびそれらの組み合わせのうちの1つから放出される、方法。
- 請求項13〜19のいずれか一項記載の方法であって、組織修復、創傷治癒を促進し、炎症を軽減する、またはそれらの組み合わせを促進する波長を有する少なくとも1秒の光に微生物感染部位を曝露することをさらに含む、方法。
- 請求項20記載の方法において、前記少なくとも1つの第2の光は、近赤外線、赤、青、および緑の範囲の波長を有する、方法。
- 請求項13〜21のいずれか一項に記載の方法であって、イオントフォレーシスをさらに含む、方法。
- 請求項13〜21のいずれか一項に記載の方法において、前記微生物感染症は、局所感染症である、方法。
- 光線力学療法のための装置であって、
光増感剤の吸収波長帯において第1の波長の光を放出するための第1の光源と、
組織修復を促進し、炎症を軽減し、創傷治癒を促進するための第2の波長で光を放出するための第2の光源と、
を含む、装置。 - 請求項24記載の装置において、前記第1の光源および前記第2の光源は、LEDである、装置。
- 請求項24または25記載の装置において、前記第2の波長が、近赤外線、赤、青、および緑の範囲にある、装置。
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