JP5950987B2 - 微生物の保存および送達のための組成物および方法 - Google Patents
微生物の保存および送達のための組成物および方法 Download PDFInfo
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- JP5950987B2 JP5950987B2 JP2014246799A JP2014246799A JP5950987B2 JP 5950987 B2 JP5950987 B2 JP 5950987B2 JP 2014246799 A JP2014246799 A JP 2014246799A JP 2014246799 A JP2014246799 A JP 2014246799A JP 5950987 B2 JP5950987 B2 JP 5950987B2
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- protective agent
- sucrose
- pharmaceutically acceptable
- glycerol
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- 238000000926 separation method Methods 0.000 description 1
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- 235000020183 skimmed milk Nutrition 0.000 description 1
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- 229940124530 sulfonamide Drugs 0.000 description 1
- 150000003456 sulfonamides Chemical class 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
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- 235000002906 tartaric acid Nutrition 0.000 description 1
- 229940095064 tartrate Drugs 0.000 description 1
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- 229960002180 tetracycline Drugs 0.000 description 1
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- 230000000699 topical effect Effects 0.000 description 1
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- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 229960003500 triclosan Drugs 0.000 description 1
- ZDPHROOEEOARMN-UHFFFAOYSA-N undecanoic acid Chemical compound CCCCCCCCCCC(O)=O ZDPHROOEEOARMN-UHFFFAOYSA-N 0.000 description 1
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Description
本発明の理解を容易にするために、いくつかの用語および句を以下に定義する。
CAUTIの予防における細菌の干渉の原理が調査されてきた。細菌の干渉は、他の種類の細菌、例えば病原体によるコロニー形成を防止するプロバイオティクスを使用する原理を指す。投与されたプロバイオティクス細菌は、例えば細菌排泄物または栄養競合によって病原性株の増殖を妨げることができるが、細菌の干渉の使用は、いかなる特定の機構にも限定されない。この方法には、優れた理論的裏付けがあり、この方法は、神経因性膀胱における症候性UTIの良好な解決法を提供し得る(Srinivasan, A., T. Karchmer, et al, Infect Control Hosp Epidemiol 27(1):38−43(2006))。
いくつかの実施形態では、本発明は、挿入前のカテーテルへの適用のために処方される、プロバイオティクス微生物の組成物を提供する。特に、特定の実施形態では、本発明は、例えば臨床医によって、投与前にプロバイオティクス生物を培養する必要なく使用されるように構成される、プロバイオティクスを含む組成物を提供する。
いくつかの実施形態では、本発明は、対象者に、プロバイオティクス微生物を投与する方法であって、該方法は、プロバイオティクス微生物、ゲル化剤、および保護剤を含む凍結乾燥調製物を提供する工程、上記の凍結乾燥調製物を水性流体に曝露し、有効量の上記のプロバイオティクス微生物を含むゲルを形成する工程、および上記の対象者を上記のゲルと接触させる工程を含む方法を提供する。いくつかの好ましい実施形態では、対象者を上記のゲルと接触させる工程は、デバイス、例えば医療デバイスをゲルと接触させ、次いで、医療デバイスを対象者と接触させる工程を含む。好ましい実施形態では、デバイスは、導尿カテーテルである。
本発明の一態様は、例えば対象者を治療するための、1つもしくは複数の構成要素を含むキットまたはトレイを提供することである。好ましい実施形態では、キットは、例えば病院、診療所、医療機関等で、医療関係者への容易な配送および医療関係者による容易な使用のために構成される。いくつかの実施形態では、本発明によるキットは、標準的なカテーテル挿入キットまたはトレイを併せて使用されるように構成されるが、他の実施形態では、本発明に従ったキットは、標準的なカテーテル挿入キットまたはトレイに代わるように構成される。好ましい実施形態では、該キットは、本発明の方法に従ったカテーテル挿入のための、全ての必要な構成要素を含む。
本発明の特定の好ましい実施形態および態様を明示し、かつさらに例示するために、以下の実施例を提供し、その範囲を制限すると解釈されないものとする。
凍結乾燥大腸菌HU2117の調製、プロトコル1
調査の目的は、HU2117の凍結乾燥のための異なる賦形剤および条件の効果を試験し、ゲル化剤を含む組成物中の凍結乾燥した細胞の有効細胞濃度および生存レベルを維持することである。
1mlのシードストックを1Lの変性EZ Rich Definedグリセロール培地に接種した、2つのフラスコ(フラスコAおよびフラスコB)の細胞を各々、250RPMの一定の振盪で、8時間、37±1℃でインキュベートして増殖させた。8時間の終わりに、フラスコAのOD600は、2.53であり、フラスコBのOD600は、1.11であった。
各試験に対して、0.5mlの再懸濁された細胞を、以下のリストから選択した1.5mlの賦形剤、および10mlの2%加熱滅菌ヒドロキシエチルセルロース(HEC)と混合した。
(a)緩衝液(賦形剤なし)
(b)2%HEC
(c)2%HEC+1.5%グリセロール
(d)2%HEC+5%トレハロース
(e)2%HEC+5%スクロース
(f)2%HEC+10%トレハロース
(g)2%HEC+10%スクロース
(h)2%HEC+5%トレハロース+1.5%グリセロール
(i)2%HEC+5%スクロース+1.5%グリセロール
(j)2%HEC+10%トレハロース+1.5%グリセロール
(k)2%HEC10%スクロース+1.5%グリセロール
細胞−賦形剤混合物を表1に記載するように凍結乾燥し、乾燥ケーキを生成した。
凍結乾燥大腸菌HU2117の調製、プロトコル2
細胞調製
1mlのシードストックを1Lの変性EZ Rich Defined グリセロール培地に接種した、2つのフラスコ(フラスコAおよびフラスコB)の細胞を各々、250RPMの一定の振盪で、8時間、37±1℃でインキュベートして増殖させた。8時間の終わりに、フラスコAのOD600は、1.89であり、フラスコBのOD600は、1.53であった。
各試験に対して、0.5mlの再懸濁された細胞を、以下のリストから選択した1.5mlの賦形剤、および10mlの1%加熱滅菌ヒドロキシエチルセルロース(HEC)と混合した。
(a)緩衝液
(b)1%HEC
(c)1%HEC+1.5%グリセロール
(d)1%HEC+5%トレハロース
(e)1%HEC+5%スクロース
(f)1%HEC+10%トレハロース
(g)1%HEC+10%スクロース
(h)1%HEC+5%トレハロース+1.5%グリセロール
(i)1%HEC+5%スクロース+1.5%グリセロール
(j)1%HEC+10%トレハロース+1.5%グリセロール
(k)1%HEC+10%スクロース+1.5%グリセロール
(l)2%HEC+10%スクロース
(m)2%HEC+10%スクロース+1.5%グリセロール
表3に記載するように、細胞混合物を凍結乾燥した。
凍結乾燥大腸菌HU2117の調製、プロトコル3
細胞調製
1Lの変性EZ Rich Defined グリセロール培地に接種した1mlのシードストックから、1つの2リットルフラスコの細胞を増殖させ、250RPMでの一定の振盪で、8時間、37±1℃でインキュベートした。8時間の終わりに、OD600は、2.2±0.03であった。
各試験に対して、0.5mlの再懸濁された細胞を、以下のリストから選択した1.5mlの賦形剤、および10mlの2%加熱滅菌ヒドロキシエチルセルロース(HEC)と混合した。
(a)緩衝液
(b)2%HEC
(c)2%HEC+5%スクロース
(d)2%HEC+10%スクロース(12ml)
(e)2%HEC+10%スクロース(10ml)
(f)2%HEC+5%スクロース+1.5%グリセロール
(g)2%HEC+10%スクロース+1.5%グリセロール
表5に記載するように、細胞混合物を凍結乾燥した。
Claims (17)
- a)大腸菌株を含むプロバイオティクス微生物、
b)ヒドロキシエチルセルロース、ヒドロキシメチルセルロース、ヒドロキシプロピルグアール、メチルセルロース、エチルセルロース、ヒドロキシプロピルセルロース、カルボキシメチルセルロースナトリウム、カルボマー、アルギン酸塩、ゼラチン、およびポロキサマーから成る群から選択される薬学的に許容されるゲル化剤、ならびに
c)グリセロール、トレハロース、およびスクロースから成る群から選択される薬学的に許容される第1の保護剤と、
を含む混合物を含む、医療用潤滑ゲルを調製するための組成物を調製するための液体組成物。 - 薬学的に許容される第2の保護剤をさらに含み、該第2の保護剤が、前記第1の保護剤とは異なり、かつ無脂肪乳固形分、トレハロース、グリセロール、ベタイン、スクロース、グルコース、ラクトース、デキストラン、ポリエチレングリコール、ソルビトール、マンニトール、ポリビニルプロピレン、グルタミン酸カリウム、グルタミン酸1ナトリウム、Tween20洗剤、Tween80洗剤、およびアミノ酸塩酸塩から成る群から選択される、請求項1に記載の組成物。
- 前記ゲル化剤が、ヒドロキシエチルセルロースである、請求項1に記載の組成物。
- 前記第1の保護剤が、スクロースである、請求項1に記載の組成物。
- 前記第1の保護剤が、スクロースであり、かつ前記第2の保護剤が、グリセロールである、請求項2に記載の組成物。
- a) i) プロバイオティクス微生物と、
ii) ヒドロキシエチルセルロース、ヒドロキシメチルセルロース、ヒドロキシプロピルグアール、メチルセルロース、エチルセルロース、ヒドロキシプロピルセルロース、カルボキシメチルセルロースナトリウム、カルボマー、アルギン酸塩、ゼラチン、およびポロキサマーから成る群から選択される薬学的に許容されるゲル化剤と、
iii) グリセロール、トレハロース、およびスクロースから成る群から選択される薬学的に許容される第1の保護剤と、
を含む混合物を水性流体中に提供する工程と、ならびに
b) 前記混合物を凍結乾燥させて、乾燥調製物を生成する工程と
を含む、医療用潤滑ゲルを調製するための凍結乾燥組成物の調製方法。 - 前記混合物が、薬学的に許容される第2の保護剤をさらに含み、該第2の保護剤が、前記第1の保護剤とは異なり、かつ無脂肪乳固形分、トレハロース、グリセロール、ベタイン、スクロース、グルコース、ラクトース、デキストラン、ポリエチレングリコール、ソルビトール、マンニトール、ポリビニルプロピレン、グルタミン酸カリウム、グルタミン酸1ナトリウム、Tween20洗剤、Tween80洗剤、およびアミノ酸塩酸塩から成る群から選択される、請求項6に記載の方法。
- 前記プロバイオティクス微生物が、大腸菌株である、請求項6に記載の方法。
- 前記ゲル化剤が、ヒドロキシエチルセルロースである、請求項6に記載の方法。
- 前記第1の保護剤が、スクロースである、請求項6に記載の方法。
- 前記第1の保護剤が、スクロースであり、かつ前記第2の保護剤が、グリセロールである、請求項7に記載の方法。
- 対象者における感染を予防または治療するために使用されるプロバイオティクス微生物で処理された医療デバイスの調製方法であって、
a)
i)大腸菌株を含むプロバイオティクス微生物、
ii)ヒドロキシエチルセルロース、ヒドロキシメチルセルロース、ヒドロキシプロピルグアール、メチルセルロース、エチルセルロース、ヒドロキシプロピルセルロース、カルボキシメチルセルロースナトリウム、カルボマー、アルギン酸塩、ゼラチン、およびポロキサマーから成る群から選択される薬学的に許容されるゲル化剤、ならびに
iii)グリセロール、トレハロース、およびスクロースから成る群から選択される薬学的に許容される第1の保護剤と、
を含む混合物を含む、医療用潤滑ゲルを提供する工程と、
b)前記医療デバイスを前記医療用潤滑ゲルと接触させ、処理されたデバイスを製造する工程と、
を含む、上記方法。 - 前記医療デバイスが、導尿カテーテルである、請求項12に記載の方法。
- a)
i)プロバイオティクス微生物、
ii)ヒドロキシエチルセルロース、ヒドロキシメチルセルロース、ヒドロキシプロピルグアール、メチルセルロース、エチルセルロース、ヒドロキシプロピルセルロース、カルボキシメチルセルロースナトリウム、カルボマー、アルギン酸塩、ゼラチン、およびポロキサマーから成る群から選択される薬学的に許容されるゲル化剤、ならびに
iii)グリセロール、トレハロース、およびスクロースから成る群から選択される薬学的に許容される第1の保護剤と、
を含む混合物を含む凍結乾燥組成物、ならびに
b)導尿カテーテル
を含む、キット。 - 前記凍結乾燥組成物が、薬学的に許容される第2の保護剤をさらに含み、該第2の保護剤が、前記第1の保護剤とは異なり、かつ無脂肪乳固形分、トレハロース、グリセロール、ベタイン、スクロース、グルコース、ラクトース、デキストラン、ポリエチレングリコール、ソルビトール、マンニトール、ポリビニルプロピレン、グルタミン酸カリウム、グルタミン酸1ナトリウム、Tween20洗剤、Tween80洗剤、およびアミノ酸塩酸塩から成る群から選択される、請求項14に記載のキット。
- 無菌水性流体の容器をさらに備える、請求項14に記載のキット。
- 前記プロバイオティクス微生物が、大腸菌株である、請求項14に記載のキット。
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US8715639B2 (en) | 2014-05-06 |
US20110020307A1 (en) | 2011-01-27 |
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US20150110759A1 (en) | 2015-04-23 |
JP5816432B2 (ja) | 2015-11-18 |
CN101795707B (zh) | 2013-03-27 |
JP2015091823A (ja) | 2015-05-14 |
CA2695419C (en) | 2015-02-17 |
US20090041727A1 (en) | 2009-02-12 |
EP2187960A4 (en) | 2010-12-08 |
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