JP6053727B2 - 抗細菌組成物 - Google Patents
抗細菌組成物 Download PDFInfo
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- JP6053727B2 JP6053727B2 JP2014165105A JP2014165105A JP6053727B2 JP 6053727 B2 JP6053727 B2 JP 6053727B2 JP 2014165105 A JP2014165105 A JP 2014165105A JP 2014165105 A JP2014165105 A JP 2014165105A JP 6053727 B2 JP6053727 B2 JP 6053727B2
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- phage
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- bactericidal composition
- lysis
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/76—Viruses; Subviral particles; Bacteriophages
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/40—Viruses, e.g. bacteriophages
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/36—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing microorganisms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/46—Deodorants or malodour counteractants, e.g. to inhibit the formation of ammonia or bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/02—Local antiseptics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/30—Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10111—Myoviridae
- C12N2795/10132—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10211—Podoviridae
- C12N2795/10232—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Zoology (AREA)
- Hematology (AREA)
- Materials Engineering (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biotechnology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Mycology (AREA)
- Agronomy & Crop Science (AREA)
- Wood Science & Technology (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Communicable Diseases (AREA)
- Dermatology (AREA)
- Oncology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicinal Preparation (AREA)
- Materials For Medical Uses (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Description
Bacterial Particles, W.H. Freeman & Co.(1963), 71−87ページ)。「リシン」として知られる溶菌酵素を単離し、そして性質決定する、多数の研究が行われてきている。リシンはまた、ムレイン加水分解酵素としても知られる。
), 71(4):1836−1842)は、黄色ブドウ球菌の異なる薬剤耐性株に対してファージKを研究した。ファージP68は、例えば、Takac M.およびBlasi U.(Antimicrob. Agents and Chemo.(2005):2934−2940)によって研究された。本発明者らは、ファージKおよび/またはP68が感染させるMRSA株の範囲が、互いに補完し、抗MRSAの組み合わせにおいて使用するのに特に適した2つの株の組み合わせをなすことを見出した。
2以上の異なるファージを用いてもよい。例えば、細菌の同じ種の異なる株の表面コーティングが異なることもある。したがって、特定の配合物が、特定の表面上の特定の種の細菌集団において、外からの溶菌を誘導しうる可能性を増加させるため、細菌の同じ種の異なる株に感染可能である2以上の異なるファージを使用することが好ましい。これはまた、特定の表面上の細菌を殺す二次的方法として、内からの溶菌もまた用いうる可能性も増加させる。
実際、ファージの複製を可能にしない細菌株に対して外からの溶菌を誘導する高い濃度、および実質的に内からの溶菌のみが起こりうる、より低い濃度の両方で、ファージKおよび/またはファージP68またはその突然変異体の組み合わせを用いてもよい。本発明者らは、殺菌配合物において、ファージKを単独で、またはファージP68と組み合わせて用いると、広範囲のMRSA株が感染し、そして殺されることが可能になることを見出した。ファージKおよびファージP68の宿主範囲は互いに補完する。
ファージKまたはファージP68を単独で、あるいは互いにまたは他のファージと組み合わせて用いてもよい。ファージKを、ファージP68を含まずに単独で、または他のファージとともに用いてもよい。
ファージKまたはP68の突然変異体は、点、欠失または付加突然変異であってもよい。元来のファージKまたはP68配列に比較して、1、2、3、4、5、6、7、8、9または10塩基を変化させてもよい。こうした突然変異体は、好ましくは、改変された宿主範囲を有する。
異なる種の細菌、および実際に異なる株の細菌に特異的なファージが、一般的に当該技術分野に知られる。例えば、組成物は、ファージKおよび/またはファージP68を含む
、MRSAを感染させうる1以上の一般的に知られるファージを含んでもよい。
本発明にしたがった組成物を感染表面に適用する工程を含む、細菌感染を治療する方法もまた提供する。本発明のさらなる側面は、細菌感染を治療するのに使用するための、本発明の第一の側面にしたがった組成物を提供する。
本発明のさらなる側面は、少なくとも1つのタイプのファージを含む包帯または創傷包帯材であって、ファージを、それが細菌の病原体となる該細菌と接触させた場合、該細菌において外からの溶菌を誘導するのに十分に高い濃度のファージが包帯または創傷包帯材上に提供されることを特徴とする、前記包帯または創傷包帯材を提供する。
また、当該技術分野に一般的に知られる技術を用いて、包帯または創傷包帯材表面上に、ファージを吸着させてもよい。
患者に包帯または創傷包帯材を適用することによって細菌を阻害するかまたは治療する方法もまた提供する。
問題のファージによる偏性溶菌性感染に感受性でない黄色ブドウ球菌細菌を、1mlあたりおよそ2x108コロニー形成単位(cfu)の濃度まで、増殖培地、LBブロス中
で増殖させた。異なる濃度のファージを、培地中に懸濁された細菌の異なるアリコットに添加して、そして37℃で一晩インキュベーションした。
ファージが感染しそして内からの溶菌を誘導してプラークを形成することが知られる細菌細胞上のプラーク形成単位(pfu)として、ファージ濃度を計算した。ファージおよび細菌細胞のpfuの計算は標準的技術である。
表1は、ファージKおよびファージP68が、ある範囲の異なる株のメチシリン耐性黄色ブドウ球菌(MRSA)において、内からの溶菌(プラーク)を誘導する能力を示す。SAI653は標準として用いられてきている。これは、ATCCより公的に入手可能であり(ATCC番号19685)、そして黄色ブドウ球菌亜種アウレウス・ローゼンバッハ(aureus Rosenbach)である。残りの株は、英国および海外の病院で患者から単離されたMRSA株である。
ファージKでは、プラーク形成を誘導不能であるファージに対して外からの溶菌を誘導するのに必要なファージKの最少濃度は、プラークpfu対細胞が5:1であることが観察された。
黄色ブドウ球菌ファージK、K*およびK*710(元来の親Kに由来する自然発生宿主範囲突然変異体)およびファージP68は、試験した大部分の黄色ブドウ球菌株の外からの溶菌を引き起こしうる。偏性溶菌性感染(内からの溶菌)に耐性である黄色ブドウ球菌突然変異体が、なお、外からの溶菌に感受性であるかどうかを調べるため、実験を行った。ファージK*710に耐性である黄色ブドウ球菌株SAI669(EMRSA−15単離体)の5つの独立の突然変異体を単離し、ファージP68に耐性である別の5つの突然変異体も同様に単離した。ファージK*710による偏性溶菌性感染に耐性であるとして単離された突然変異体はすべて、ファージKおよびK*による偏性溶菌性感染にも耐性を示した。K/K*/K*710による偏性溶菌性感染に耐性である突然変異体は、ファージP68による偏性溶菌性感染に感受性であり、そして逆に、ファージP68による偏
性溶菌性感染に耐性である突然変異体は、なお、K/K*/K*710による偏性溶菌性感染に感受性であった。個々のファージ耐性突然変異体各々を、外からの溶菌に対する感受性に関して試験した。Lブロス寒天(1.5% w/v)のプレート上に、3mlのLブロス上層寒天(0.7% w/v)中の黄色ブドウ球菌株菌叢を接種し、そして次いで、20μlの10倍連続希釈の各ファージ(最初の濃度が〜5x109プラーク形成単位/ml)を上層寒天上にスポッティングし、その後、37℃で20時間インキュベーションすることによって、この試験を行った。細菌叢中、高濃度のファージで透明になる領域があるが、低濃度のファージでは個々のプラークが存在しない場合、外からの溶菌を示すと解釈された。
Claims (16)
- キャリアーおよび2以上のファージを含む、表面を殺菌するための殺菌組成物であって、ファージはファージKまたはその突然変異体およびファージP68またはその突然変異体を含み、ファージKまたはその突然変異体およびファージP68またはその突然変異体が細菌の病原体である黄色ブドウ球菌が、殺菌しようとする表面上に存在する場合、該黄色ブドウ球菌において外からの溶菌(lysis−from−without)を誘導するのに十分に高い濃度の両方のファージが組成物中に提供されることを特徴とし、ファージKまたはその突然変異体の濃度はpfuファージ:cfu細菌が少なくとも5:1であって外からの溶菌を誘導する、前記組成物。
- ファージ濃度は、pfuファージ:cfu細菌が少なくとも5:1である、請求項1に記載の殺菌組成物。
- 哺乳動物皮膚へ適用するために局所適用するものであるか、または表面用のスプレーまたは液体洗浄剤である、請求項1に記載の殺菌組成物。
- 皮膚が鼻腔内にあるか、またはヒトの手の皮膚である、請求項3記載の殺菌組成物。
- クリーム、ローション、軟膏、ペースト、ジェル、泡(foam)、または手洗い剤の形である、請求項3または請求項4記載の殺菌組成物。
- キャリアーがラノリンまたはパラフィンを含む、請求項3または請求項4記載の殺菌組成物。
- 殺菌組成物中のファージが、異なった種の細菌に対する特異的な病原体であるファージを更に含み、該細菌が、ヘリコバクター属(Helicobacter)、クレブシエラ属(Klebsiella)、リステリア属(Listeria)、マイコバクテリウム属(Mycobacterium)、エシェリキア属(Escherichia)、髄膜炎菌属(Meningococcus)、カンピロバクター属(Campylobacter)、連鎖球菌属(Streptococcus)、腸球菌属(Enterococcus)、赤痢菌属(Shigella)、シュードモナス属(Pseudomonas)、バークホルデリア属(Burkholderia)、クロストリジウム属(Clostridium)、レジオネラ属(Legionella)、アセチノバクター属(Acetinobacter)、またはサルモネラ属(Salmonella)より選択される、請求項1に記載の殺菌組成物。
- 突然変異体は、点、欠失または付加突然変異であり、元来のファージKまたはP68配列と比較して1〜10塩基が変化している、請求項1に記載の殺菌組成物。
- 水、保存剤、活性界面剤、乳化剤、酸化防止剤、または溶媒の1以上を更に含む、請求項1、請求項4、または請求項8に記載の殺菌組成物。
- 請求項1〜9のいずれか一項記載の殺菌組成物を含む、包帯または創傷包帯材。
- 表面上の細菌を殺すための薬剤を製造するための、請求項1〜9のいずれか一項記載の殺菌組成物の使用。
- 表面が哺乳動物皮膚である、請求項11に記載の使用。
- 表面が哺乳動物の鼻腔である、請求項11に記載の使用。
- 表面上の細菌を殺す方法であって、請求項1〜9のいずれか一項記載の殺菌組成物を前記表面に適用する工程を含み、前記表面が病院における医療機器、寝具類、家具、壁または床である、上記方法。
- 細菌感染の治療のための薬剤を製造するための、請求項1〜9のいずれか一項記載の殺菌組成物の使用。
- 細菌感染が感染した表面である、請求項15に記載の使用。
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