JP6768705B2 - 創傷被覆材用の組成物 - Google Patents
創傷被覆材用の組成物 Download PDFInfo
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- JP6768705B2 JP6768705B2 JP2017557505A JP2017557505A JP6768705B2 JP 6768705 B2 JP6768705 B2 JP 6768705B2 JP 2017557505 A JP2017557505 A JP 2017557505A JP 2017557505 A JP2017557505 A JP 2017557505A JP 6768705 B2 JP6768705 B2 JP 6768705B2
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- acid
- component
- tribasic
- chitosan
- solubilized
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Description
(a)繊維状の第1の成分の少なくとも一部分を、少なくとも1種の三塩基酸及び少なくとも1種の可溶化酸を含む混合物で被覆する工程と、及び/または
(b)第1の成分の少なくとも一部分に、少なくとも1種の三塩基酸及び少なくとも1種の可溶化酸を含む混合物を吸収させる工程とを含む。
(a)水及び/または溶剤に三塩基酸を混合して三塩基酸溶液を得る工程と、
(b)水及び/または溶剤に可溶化酸を混合することにより調製した可溶化酸溶液に、三塩基酸溶液を混合して、混合酸性溶液を得る工程と、
(c)任意に、溶剤に混合酸性溶液を混合する工程と、
(d)工程(c)で得られた溶液に繊維状の第1の成分を加える工程とを含む。
一般的な方法1:第1の成分と三塩基酸
三塩基酸(例えばクエン酸)粉末を脱イオン水に溶解し、次に非水性の溶剤(例えばIPA)と混合した。第1の成分は、典型的には不織繊維の形態であり、三塩基酸溶液中に配置して、溶液を吸収させた。次に溶液を加熱乾燥を用いて乾燥し、三塩基酸で被覆した固体のキトサン、キチンまたはこれらの誘導体を残存させた。
三塩基酸(例えばクエン酸)粉末を脱イオン水に溶解し、次に可溶化酸(例えば乳酸)溶液と混合した。これを続いて非水性溶剤(例えばIPA)と混合した。第1の成分は、典型的には不織繊維の形態であり、混合酸性溶液中に配置して、溶液を吸収させた。次に溶液を加熱乾燥を用いて乾燥し、三塩基酸と可溶化酸との混合物で被覆した固体のキトサン、キチンまたはこれらの誘導体を残存させた。
以下、本発明に従って調製された組成物の実施例である。
実施例14(参考例):本発明の組成物は、0.6gのクエン酸を入れた8gの脱イオン水の中に、公称2gの100%キトサン不織繊維を含んで調製する。組成物は、非網目状の(non-reticulated)ポリウレタンフォームの表面上に被覆し、乾燥させた。本発明の乾燥した組成物を含むフォームは、次にポリウレタンフォームに接着剤で結合され、結合されたポリウレタンフィルム層より遠位に、創傷接触層が形成された。
さまざまな微生物のバイオフィルムに対する抗菌効能を測定するために、MBEC (最小バイオフィルム形成阻止濃度、Minimum Biofilm Eradication Concentration)アッセイを使用した。
緑膿菌ATCC13359
スタフィロコッカス−ヘモリチカス
MRSA308。
対照(Control):リン酸緩衝食塩水
試料(Sample)A:銀入り(公称1%)100%キトサン不織繊維と乳酸より得られた、公称25%一塩基酸組成物
試料B:銀入り(公称1%)カルボキシメチル化セルロース不織繊維(Aquacel Ag[登録商標])
試料C:乳酸入り100%キトサン不織繊維より得られた、公称25%一塩基酸組成物
試料D:酢酸入り100%キトサン不織繊維より得られた、公称25%一塩基酸組成物
試料E:クエン酸入り100%キトサン不織繊維より得られた、公称25%三塩基酸組成物。
各微生物の24時間培養物は、トリプトンソーヤ寒天培地(Tryptone Soya Agar (TSA))プレートまたは ブレインハートインフュージョン寒天培地(Brain Heart Infusion Agar (BHIA))プレートのいずれかから採取され、20mlのトリプトンソーヤブイヨン培地(Tryptone Soya Broth (TSB))または20mlのブレインハートインフュージョンブイヨン培地(Brain Heart Infusion Broth (BHIB))のいずれかの中に懸濁させた。その結果得られたバクテリア懸濁液を希釈して、108cfuml-1のバクテリア濃度に対応する初期OD590 = 0.10 ± 0.03を得た。この初期の種菌は、次第に低くなる細菌量(すなわち107, 106, 105, 104, 103 cfuml-1)が表れるように、6回連続して希釈された。各生物の開始時のバクテリア濃度は、典型的には 1 x 108± 5 x 107cfuml-1だった。
各微生物のバイオフィルムは、マイクロタイタープレートのピンふた突起(pin lid projections)上で48時間、37℃、50rmpで育成した。48時間後、ピンふたを取り外して、リン酸緩衝食塩水で短時間洗浄してプランクトン状のバクテリアを除去し、続いて薬剤チャレンジプレート(agent challenge plate)中に24時間配置した。創傷被覆材用のチャレンジプレートの調製のために、無菌のはさみを使用して1cm2 の断片が切り取られ、マイクロタイタープレートの指定されたウェル内に配置された。顆粒の試験薬剤用のチャレンジプレートは、マイクロタイタープレートのウェル内に各顆粒製剤を30mg ± 3mgに量り分けることにより調製した。次に創傷被覆材及び顆粒を150μlのPBSにより活性化した。処理に続いて、残った付着バクテリアを回収するために、ピンふた突起をPBS内で2回洗浄し、次に200μlの中和剤中に移し、音波水浴(sonic water-bath)中に5分間配置した。連続希釈が得られた回収液(recovery broth)について実行され、ドロッププレート(drop plate)が回収したバクテリアの量を計るために使用された。全ての試料は、特に明記しない限り、三回試験された(tested in triplicate)。
スタフィロコッカス−ヘモリチカスNCTC11042
緑膿菌ATCC10434
メチシリン耐性黄色ブドウ球菌308。
対照:リン酸緩衝食塩水(PBS)
試料F:15%乳酸及び3%クエン酸を含む100%キトサン不織繊維。これはキトサン1.35g、乳酸0.2g及びクエン酸0.04g(実施例8)と同等である
試料G:15%乳酸及び25%クエン酸を含む100%キトサン不織繊維。これはキトサン1.35g、乳酸0.2g及びクエン酸0.34g(実施例9)と同等である
試料H:20%乳酸及び60%クエン酸を含む100%キトサン不織繊維。これはキトサン1.35g、乳酸0.27g及びクエン酸0.81g(実施例10)と同等である
試料I:3%クエン酸を含む100%キトサン不織繊維。これはキトサン1.35g及びクエン酸0.04g(参考実施例2)と同等である
試料J:25%クエン酸を含む100%キトサン不織繊維。これはキトサン1.35g及びクエン酸0.34g(参考実施例3)と同等である
試料K:60%クエン酸を含む100%キトサン不織繊維。これはキトサン1.35g及びクエン酸0.81g(参考実施例4)と同等である。
試験バクテリアの24時間培養物は、トリプトンソーヤ寒天培地(TSA)プレートから無菌綿棒を使用して採取され、20mlのトリプトンソーヤブイヨン培地(TSB)の中に再懸濁させた。バクテリア懸濁液を希釈して、108± 5 x107cfuml-1のバクテリア濃度に対応する、初期OD590 = 0.10 ± 0.03を得た。これをさらにTSB内で希釈して、試験材を含むCDCリアクターのための種菌として使用した。CDCリアクターを、バイオフィルムの成長を促すために50rpmで振盪しながら、37℃で48時間培養した。
48時間後、試験材をCDCリアクターから取り外し、プランクトン状のバクテリアを除去するために無菌PBS内で3回洗浄した。次に洗浄した試験材を2枚の1.5 cm2 創傷被覆材の間に試験材を挟むことにより処理した。試験に先立って、各1.5 cm2 断片に350μlPBS(75%飽和)を添加することにより、被覆材を活性化した。対照材は、緑膿菌用に2mlのPBSの中(あるいはスタフィロコッカス−ヘモリチカス及びMRSAの場合は2mlPBS+0.1%TSBの中)に浸水した。全ての試料を3回試験した。微生物は、24時間の処理後に試験材から回収し、連続希釈とドロッププレートを実行して量を計った。
2種のバクテリア種に対する6種類の創傷被覆材のバイオフィルム除去能力を測定するために、CDCリアクター方法を使用した。
スタフィロコッカス−ヘモリチカスNCTC8325
緑膿菌ATCC10434。
対照:リン酸緩衝食塩水(PBS)
試料L:25%乳酸及び30%クエン酸を含む100%キトサン不織繊維。これはキトサン1.35g、乳酸0.34g及びクエン酸0.41g(実施例11)と同等である
試料M:25%乳酸及び40%クエン酸を含む100%キトサン不織繊維。これはキトサン1.35g、乳酸0.34g及びクエン酸0.54g(実施例12)と同等である
試料N:30%乳酸及び40%クエン酸を含む100%キトサン不織繊維。これはキトサン1.35g、乳酸0.41g及びクエン酸0.54g(実施例13)と同等である
試料O:100%キトサン不織繊維
試料P:25%乳酸を含む55%キトサン繊維/45%ビスコース不織繊維。これはキトサン0.74g、ビスコース0.61g及びクエン酸0.34gと同等である
試料Q:イオン状態の銀含有物(ionic silver-containing)を含む不織カルボキシメチル化セルロース、抗バイオフィルム製剤。
各微生物の24時間培養物は、トリプトンソーヤ寒天培地(TSA)プレートから採取され、20mlのトリプトンソーヤブイヨン培地(TSB)の中に再懸濁させた。その結果得られたバクテリア懸濁液を希釈して、108 ± 5 x107cfuml-1のバクテリア濃度に対応する、初期OD590 = 0.10 ± 0.03を得た。これをさらにTSB中で約107 cfuml-1まで希釈し、CDCリアクター用の初期種菌として使用した。CDCリアクターを、バイオフィルムの成長を促すために50rpmで振盪しながら、37℃で24時間及び72時間培養した。
24時間後及び72時間後、試験材をCDCリアクターから取り外して、プランクトン状のバクテリアを除去するために、無菌リン酸緩衝食塩水(PBS)中で3回洗浄した。次に洗浄した試験材を2枚の創傷被覆材の間に試験材を挟むことにより処理した。試験に先立って、1%のTSBを含む350μlのPBSを添加することにより、被覆材を活性化した。対照の試験材は、1%のTSBを含む2mlのPBS中に浸水した。24時間の処理時間に続いて、試験材は2mlの中和剤中に配置され、15分間超音波処理されて、残った付着バクテリアを再生した。連続希釈が得られた回収液(recovery broth)について実行され、ドロッププレート(drop plate)が回収したバクテリアの量を計るために使用された。全ての試料について試験は3回行われた。
Claims (18)
- キトサン、キチン、キトサン誘導体、キチン誘導体、及びこれらの任意の組み合わせからなる群から選択された繊維状の第1の成分と、少なくとも1種の三塩基酸と、少なくとも1種の可溶化酸とを含み、前記少なくとも1種の三塩基酸と前記少なくとも1種の可溶化酸とは、前記第1の成分の少なくとも一部分の上に被覆され、及び/または前記第1の成分の少なくとも一部分の中に吸収されている組成物。
- 前記第1の成分は非抗菌であり、該非抗菌は24時間以内にlog4のバクテリア殺し率を実証する薬剤または物質を指す請求項1に記載の組成物。
- 前記少なくとも1種の三塩基酸に対する前記第1の成分の比率は少なくとも2:1であり、及び/または前記少なくとも1種の可溶化酸に対する前記少なくとも1種の三塩基酸の比率は少なくとも1:1である請求項1または2に記載の組成物。
- 前記第1の成分はキトサンであり、及び/または前記三塩基酸はクエン酸である請求項1乃至3のいずれか1項に記載の組成物。
- 前記第1の成分は、70%を超える脱アセチル化度を有し、及び/または前記第1の成分は、1%酢酸溶液中で150cpsを超える粘度を有する請求項4に記載の組成物。
- 前記三塩基酸は、前記第1の成分の約2から75%の量が存在する請求項1乃至5のいずれか1項に記載の組成物。
- 前記三塩基酸は、前記第1の成分の約25から60%の量が存在する請求項6に記載の組成物。
- 前記可溶化酸は一塩基酸である請求項1乃至7のいずれか1項に記載の組成物。
- 前記一塩基酸は、乳酸、ギ酸、酢酸、塩酸、コハク酸及びこれらの混合からなる群から選択される請求項8に記載の組成物。
- 前記可溶化酸と前記三塩基酸とは、前記第1の成分に接触する前に混合され、及び/または前記可溶化酸及び前記三塩基酸は、前記第1の成分の上に被覆されている請求項1乃至9のいずれか1項に記載の組成物。
- 前記可溶化酸は、前記第1の成分の2から50%の量が存在する請求項1乃至10のいずれか1項に記載の組成物。
- 前記可溶化酸は、前記第1の成分の15から20%の量が存在する請求項11に記載の組成物。
- 前記三塩基酸及び/または前記可溶化酸は、担体材料の上に被覆されている請求項1乃至12のいずれか1項に記載の組成物。
- 抗菌剤、医薬品、キレート剤、湿潤剤、成長因子、サイトカイン、MMP(マトリックスメタロプロテアーゼ)及びエラスターゼのような治癒を遅延させる物質を吸収する薬剤、カルシウム、ビタミンK、フィブリノゲン、トロンビン、第VII因子、第VIII因子、粘土、酸化再生セルロース、ゼラチン及びコラーゲンからなる群から選択された付加的な成分をさらに含む請求項1乃至13のいずれかに1項に記載の組成物。
- 請求項1乃至14のいずれか1項に記載の組成物を含む創傷被覆材。
- 治療薬として使用する請求項1乃至14のいずれか1項に記載の組成物。
- 創傷の治療に使用する、またはバイオフィルムにおけるバクテリアの破壊と殺しに使用する、またはバイオフィルムの形成の防止に使用する請求項1乃至14のいずれか1項に記載の組成物。
- キトサン、キチン、キトサン誘導体、キチン誘導体、及びこれらの組み合わせからなる群から選択された繊維状の第1の成分と、少なくとも1種の三塩基酸と、少なくとも1種の可溶化酸とを含む組成物の製造方法であって、
a.前記繊維状の第1の成分の少なくとも一部分を、前記少なくとも1種の三塩基酸と前記少なくとも1種の可溶化酸とを含む混合物で被覆する工程、及び/または
b.前記第1の成分の少なくとも一部分に、前記少なくとも1種の三塩基酸と前記少なくとも1種の可溶化酸とを含む混合物を吸収させる工程、
を含む方法。
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WO2008143889A1 (en) | 2007-05-14 | 2008-11-27 | Research Foundation Of State University Of New York | Induction of a physiological dispersion response in bacterial cells in a biofilm |
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GB0817796D0 (en) | 2008-09-29 | 2008-11-05 | Convatec Inc | wound dressing |
GB201020236D0 (en) | 2010-11-30 | 2011-01-12 | Convatec Technologies Inc | A composition for detecting biofilms on viable tissues |
ES2748519T3 (es) | 2010-12-08 | 2020-03-17 | Convatec Technologies Inc | Accesorio de sistema de exudado de heridas |
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GB2497406A (en) | 2011-11-29 | 2013-06-12 | Webtec Converting Llc | Dressing with a perforated binder layer |
MX2015007771A (es) | 2012-12-20 | 2015-09-04 | Convatec Technologies Inc | Procesamiento de fibras celulosicas quimicamente modificadas. |
AU2017243601A1 (en) | 2016-03-30 | 2018-11-22 | Acib Gmbh | Detecting microbial infection in wounds |
UY37178A (es) | 2016-03-30 | 2017-10-31 | Convatec Technologies Inc | Detección de infección microbiana en heridas |
ES2882336T3 (es) | 2016-07-08 | 2021-12-01 | Convatec Technologies Inc | Sistema flexible de presión negativa |
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US11541105B2 (en) | 2018-06-01 | 2023-01-03 | The Research Foundation For The State University Of New York | Compositions and methods for disrupting biofilm formation and maintenance |
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US11771819B2 (en) | 2019-12-27 | 2023-10-03 | Convatec Limited | Low profile filter devices suitable for use in negative pressure wound therapy systems |
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