JP2017200911A - Antimicrobial agent - Google Patents
Antimicrobial agent Download PDFInfo
- Publication number
- JP2017200911A JP2017200911A JP2017088146A JP2017088146A JP2017200911A JP 2017200911 A JP2017200911 A JP 2017200911A JP 2017088146 A JP2017088146 A JP 2017088146A JP 2017088146 A JP2017088146 A JP 2017088146A JP 2017200911 A JP2017200911 A JP 2017200911A
- Authority
- JP
- Japan
- Prior art keywords
- propionibacterium
- pharmaceutical composition
- acne
- bacteria
- methyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
【課題】本発明の目的は、主として、１−シクロプロピル−８−メチル−７−［５−メチル−６−（メチルアミノ）−３−ピリジル］−４−オキソ−１，４−ジヒドロ−３−キノリンカルボン酸及び／又はその医薬上許容される塩を含有する医薬組成物であって、プロピオニバクテリウム属の細菌に対して増殖抑制作用を有することを特徴とする医薬組成物を提供することにある。【解決手段】本発明としては、例えば、１−シクロプロピル−８−メチル−７−［５−メチル−６−（メチルアミノ）−３−ピリジル］−４−オキソ−１，４−ジヒドロ−３−キノリンカルボン酸及び／又はその医薬上許容される塩を有効成分として含有する、プロピオニバクテリウム属の細菌に対して増殖抑制作用を有することを特徴とする医薬組成物を挙げることができる。【選択図】なしPROBLEM TO BE SOLVED: To provide mainly 1-cyclopropyl-8-methyl-7- [5-methyl-6- (methylamino) -3-pyridyl] -4-oxo-1,4-dihydro-3. -Providing a pharmaceutical composition containing a quinoline carboxylic acid and / or a pharmaceutically acceptable salt thereof, which has a growth inhibitory effect on bacteria of the genus Propionibacterium. There is. According to the present invention, for example, 1-cyclopropyl-8-methyl-7- [5-methyl-6- (methylamino) -3-pyridyl] -4-oxo-1,4-dihydro-3. -A pharmaceutical composition comprising a quinoline carboxylic acid and / or a pharmaceutically acceptable salt thereof as an active ingredient and having a growth inhibitory effect on a bacterium of the genus Propionibacterium can be mentioned. [Selection diagram] None
Description
本発明は、１−シクロプロピル−８−メチル−７−［５−メチル−６−（メチルアミノ）−３−ピリジル］−４−オキソ−１，４−ジヒドロ−３−キノリンカルボン酸（以下、「化合物Ａ」という）及び／又はその医薬上許容される塩を有効成分として含有する、プロピオニバクテリウム属の細菌に対して増殖抑制作用を有することを特徴とする医薬組成物に関するものである。
The present invention relates to 1-cyclopropyl-8-methyl-7- [5-methyl-6- (methylamino) -3-pyridyl] -4-oxo-1,4-dihydro-3-quinolinecarboxylic acid (hereinafter, The present invention relates to a pharmaceutical composition characterized by having a growth-inhibiting action against Propionibacterium bacteria, which contains “Compound A”) and / or a pharmaceutically acceptable salt thereof as an active ingredient. .
プロピオニバクテリウム属に属するプロピオニバクテリウム・アクネスは、尋常性ざ瘡等の化膿性炎症を伴うざ瘡の原因菌であり、毛包脂腺管内で増殖することにより疾患が発症する。
従来、化膿性炎症を伴うざ瘡の主な治療法としては、軽度から中等度ではナジフロキサシン等の外用抗菌剤、中等度から重度ではミノサイクリン、ロキシスロマイシン等の経口抗菌剤が繁用されている。
外用抗菌剤に関して、新たな医薬品として、化合物Ａを有効成分として含有する製剤の開発も行われている（例えば、特許文献１、特許文献２を参照）。
Propionibacterium acnes belonging to the genus Propionibacterium is a causative bacterium of acne with purulent inflammation such as acne vulgaris, and the disease develops when it grows in the hair follicle gland duct.
Conventionally, as a main treatment for acne with purulent inflammation, external antibacterial agents such as nadifloxacin have been frequently used for mild to moderate, and oral antibacterial agents such as minocycline and roxithromycin have been frequently used for moderate to severe .
Regarding external antibacterial agents, as a new pharmaceutical, a preparation containing Compound A as an active ingredient has also been developed (see, for example, Patent Document 1 and Patent Document 2).
本発明の目的は、主として、化合物Ａを含有する医薬組成物であって、プロピオニバクテリウム属の細菌に対して増殖抑制作用を有することを特徴とする医薬組成物を提供することにある。
An object of the present invention is to provide a pharmaceutical composition mainly containing Compound A, which has a growth inhibitory action against Propionibacterium bacteria.
本発明者らは、鋭意検討した結果、化合物Ａを有効成分として用いることにより、上記の課題が解決できることを見出し、本発明を完成するに至った。 As a result of intensive studies, the present inventors have found that the above problems can be solved by using Compound A as an active ingredient, and have completed the present invention.
本発明としては、例えば、以下のものを挙げることができる。
（１）化合物Ａ及び／又はその医薬上許容される塩を有効成分として含有する、プロピオニバクテリウム属の細菌に対して増殖抑制作用を有することを特徴とする医薬組成物（以下、「本発明組成物」という）。
（２）プロピオニバクテリウム属の細菌が、キノロン系抗生物質に対して感受性を有する菌株である、上記１に記載の本発明組成物。
（３）プロピオニバクテリウム属の細菌が、プロピオニバクテリウム・アクネスである、上記１又は２に記載の本発明組成物。
（４）プロピオニバクテリウム属の細菌への感染により生じる疾患の治療及び／又は予防に用いることを特徴とする、上記１〜３のいずれかに記載の本発明組成物。
（５）プロピオニバクテリウム属の細菌への感染により生じる疾患が、プロピオニバクテリウム・アクネスに感染することにより生じる、化膿性炎症を伴うざ瘡である、上記４に記載の医薬組成物。
（６）化膿性炎症を伴うざ瘡が、尋常性ざ瘡、新生児ざ瘡又は集簇性ざ瘡である、上記５に記載の本発明組成物。
Examples of the present invention include the following.
(1) A pharmaceutical composition containing a compound A and / or a pharmaceutically acceptable salt thereof as an active ingredient and having a growth-inhibiting action against bacteria of the genus Propionibacterium (hereinafter referred to as “this” Inventive composition ").
(2) The composition of the present invention described in 1 above, wherein the Propionibacterium bacterium is a strain having sensitivity to quinolone antibiotics.
(3) The composition of the present invention as described in 1 or 2 above, wherein the Propionibacterium bacterium is Propionibacterium acnes.
(4) The composition of the present invention as described in any one of 1 to 3 above, which is used for treatment and / or prevention of a disease caused by infection with bacteria of the genus Propionibacterium.
(5) The pharmaceutical composition according to 4 above, wherein the disease caused by infection with Propionibacterium spp. Is acne with purulent inflammation caused by infection with Propionibacterium acnes.
(6) The composition of the present invention as described in 5 above, wherein the acne with purulent inflammation is acne vulgaris, neonatal acne or collected acne.
化合物Ａは、キノロン系合成抗菌化合物に分類され、細菌のＤＮＡ複製に関与するＤＮＡジャイレース及びトポイソメラーゼＩＶを阻害して抗菌作用を発揮する。
また、化合物Ａは、グラム陽性菌、グラム陰性菌、嫌気性菌、クラミジア及び薬剤耐性グラム陽性菌に対して幅広い抗菌スペクトルと強い抗菌活性を有している。
Compound A is classified as a quinolone synthetic antibacterial compound, and exhibits antibacterial action by inhibiting DNA gyrase and topoisomerase IV involved in bacterial DNA replication.
Compound A has a broad antibacterial spectrum and strong antibacterial activity against gram positive bacteria, gram negative bacteria, anaerobic bacteria, chlamydia and drug-resistant gram positive bacteria.
化合物Ａは、ＷＯ９９／５１５８８に記載の方法により合成することができる。 Compound A can be synthesized by the method described in WO99 / 51588.
化合物Ａの医薬上許容される塩としては、通常知られているアミノ基等の塩基性基又はヒドロキシル基若しくはカルボキシル基等の酸性基における塩を挙げることができる。 Examples of the pharmaceutically acceptable salt of Compound A include salts that are generally known in basic groups such as amino groups or acidic groups such as hydroxyl groups or carboxyl groups.
塩基性基における塩としては、例えば、塩酸、臭化水素酸、硫酸等の鉱酸との塩；酒石酸、ギ酸、フマル酸、マレイン酸、リンゴ酸、クエン酸等の有機カルボン酸との塩；並びにメタンスルホン酸、ベンゼンスルホン酸、ｐ−トルエンスルホン酸、メシチレンスルホン酸及びナフタレンスルホン酸などのスルホン酸との塩を挙げることができる。 Examples of the salt in the basic group include salts with mineral acids such as hydrochloric acid, hydrobromic acid and sulfuric acid; salts with organic carboxylic acids such as tartaric acid, formic acid, fumaric acid, maleic acid, malic acid and citric acid; And salts with sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, mesitylenesulfonic acid and naphthalenesulfonic acid.
酸性基における塩としては、例えば、ナトリウム、カリウム等のアルカリ金属との塩；カルシウム、マグネシウム等のアルカリ土類金属との塩；アンモニウム塩；並びにリジン、アルギニン、オルニチン等のアミノ酸、トリメチルアミン、トリエチルアミン、トリブチルアミン、ピリジン、Ｎ，Ｎ−ジメチルアニリン、Ｎ−メチルピペリジン、Ｎ−メチルモルホリン、ジエチルアミン、ジシクロヘキシルアミン、プロカイン、ジベンジルアミン、Ｎ−ベンジル−β−フェネチルアミン、１−エフェナミン並びにＮ，Ｎ'−ジベンジルエチレンジアミン等の含窒素有機塩基との塩を挙げることができる。 Examples of the salt in the acidic group include salts with alkali metals such as sodium and potassium; salts with alkaline earth metals such as calcium and magnesium; ammonium salts; and amino acids such as lysine, arginine and ornithine, trimethylamine, triethylamine, Tributylamine, pyridine, N, N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, diethylamine, dicyclohexylamine, procaine, dibenzylamine, N-benzyl-β-phenethylamine, 1-ephenamine and N, N′- Mention may be made of salts with nitrogen-containing organic bases such as dibenzylethylenediamine.
本発明組成物は、プロピオニバクテリウム属の細菌に対して増殖抑制作用を有するものであり、プロピオニバクテリウム属の細菌への感染により生じる疾患の治療及び／又は予防に用いることができる。 The composition of the present invention has a growth inhibitory action against Propionibacterium spp. And can be used for treatment and / or prevention of diseases caused by infection with Propionibacterium spp.
本発明に係る「増殖抑制作用」とは、細菌の増殖を止める作用（静菌作用）を意味する。
静菌作用とは、細菌の増殖を抑制する作用を意味し、細菌を死滅させる殺菌作用とは明確に区別される。
殺菌作用とは、一定濃度以上の薬剤を被験菌に作用させることで、一定時間後に接種時の生菌数を１／１０００以下に減少させる作用を意味する。
The “growth inhibitory action” according to the present invention means an action to stop bacterial growth (bacteriostatic action).
The bacteriostatic action means an action that suppresses the growth of bacteria, and is clearly distinguished from a bactericidal action that kills bacteria.
A bactericidal action means the effect | action which reduces the number of living microbes at the time of inoculation to 1/1000 or less after fixed time by making a test | inspection chemical | medical agent act on a test microbe.
増殖抑制作用は、有効成分がＰＡＥを有するか否かを一つの指標として確認することができる。 The growth inhibitory action can be confirmed as one index whether or not the active ingredient has PAE.
ＰＡＥとは、「ある抗菌薬が細菌に短時間接触した後に持続してみられる増殖抑制効果」と定義される。即ち、ＰＡＥを有する抗菌薬は、血中若しくは組織内からその薬剤が消失した後も、細菌の増殖をある一定期間抑制できることを意味する。 PAE is defined as “a growth-suppressing effect that is sustained after a certain antibacterial drug is in contact with bacteria for a short time”. That is, an antibacterial drug having PAE means that the growth of bacteria can be suppressed for a certain period even after the drug disappears from blood or tissue.
本発明に係る「プロピオニバクテリウム属の細菌」としては、例えば、プロピオニバクテリウム・アクネス、プロピオニバクテリウム・アビダム、プロピオニバクテリウム・リンフォフィラム、プロピオニバクテリウム・グラヌローサム、プロピオニバクテリウム・ソエニイ又はプロピオニバクテリウム・プロピオニカムを挙げることができる。それらの中で、特にプロピオニバクテリウム・アクネスが好ましい。 Examples of the “Propionibacterium bacterium” according to the present invention include, for example, Propionibacterium acnes, Propionibacterium abidam, Propionibacterium lymphophilum, Propionibacterium granulosum, Propionibacterium Mention may be made of Soeniy or Propionibacterium propionicam. Among them, Propionibacterium acnes is particularly preferable.
本発明に係る「プロピオニバクテリウム属の細菌への感染により生じる疾患」としては、例えば、プロピオニバクテリウム・アクネスに感染することにより生じる、化膿性炎症を伴うざ瘡を挙げることができる。 Examples of the “disease caused by infection with bacteria of the genus Propionibacterium” according to the present invention include acne accompanied by purulent inflammation caused by infection with Propionibacterium acnes.
化膿性炎症を伴うざ瘡としては、例えば、尋常性ざ瘡、新生児ざ瘡、集簇性ざ瘡を挙げることができる。それらの中で、特に尋常性ざ瘡が好ましい。 Examples of acne with purulent inflammation include acne vulgaris, neonatal acne, and confluent acne. Among them, acne vulgaris is particularly preferable.
本発明組成物の剤型は、特に限定されないが、例えば、軟膏剤、ゲル剤、クリーム剤、乳剤、粘着テープ剤、ローション剤を挙げることができる。 The dosage form of the composition of the present invention is not particularly limited, and examples thereof include ointments, gels, creams, emulsions, adhesive tapes, and lotions.
本発明組成物は、当業者に自明な方法により、構成成分を適宜混合し調製することができる。 The composition of the present invention can be prepared by appropriately mixing the components by a method obvious to those skilled in the art.
本発明組成物における、化合物Ａ及び／又はその医薬上許容される塩の含有量は、治療効果を発揮する量であれば特に限定されないが、例えば、製剤中に０．０１〜２０重量％の範囲内が適当であり、０．１〜５重量％の範囲内が好ましい。 The content of Compound A and / or a pharmaceutically acceptable salt thereof in the composition of the present invention is not particularly limited as long as it is an amount that exhibits a therapeutic effect. The range is suitable, and the range of 0.1 to 5% by weight is preferable.
本発明組成物の投与量は、患者の年齢、体重及び症状に応じて適宜選択されるが、通常、薬効を発揮し得る量として、外用剤として投与される場合には、１日３０〜２０００ｍｇを経皮投与すればよい。
The dose of the composition of the present invention is appropriately selected according to the age, weight and symptoms of the patient, but usually 30 to 2000 mg per day when administered as an external preparation as an amount capable of exerting a medicinal effect. May be administered transdermally.
以下に、試験例を掲げて、本発明を更に詳しく説明するが、本発明は実施例に示される範囲に限定されるものではない。 Hereinafter, the present invention will be described in more detail with reference to test examples, but the present invention is not limited to the scope shown in the examples.
[試験例１] 各種抗菌薬のＰＡＥの評価
１）試験物質
本試験では、以下の抗菌薬を試験物質として用いた。
・化合物Ａ（力価：９９．７％）
・ナジフロキサシン（力価：９９．９％）
・レボフロキサシン（力価：９６．８％）
２）使用菌株
２０１２年〜２０１３年に尋常性ざ瘡患者より分離したプロピオニバクテリウム・アクネスを用いた。
使用菌株に対する各試験物質の最小発育阻止濃度（Ｍｉｎｉｍｕｍ ｉｎｈｉｂｉｔｏｒｙ ｃｏｎｃｅｎｔｒａｔｉｏｎ：ＭＩＣ）（μｇ／ｍＬ）は、以下の通りであり、各試験物質に対して感受性を有することを確認した。
なお、ＭＩＣの測定は、Ｃｌｉｎｉｃａｌ ａｎｄ Ｌａｂｏｒａｔｏｒｙ Ｓｔａｎｄａｒｄｓ Ｉｎｓｔｉｔｕｔｅの推奨する微量液体希釈法に準じて行った。
・化合物Ａ（０．０３）
・ナジフロキサシン（０．１２５）
・レボフロキサシン（０．５）
３）培地の調製
（１）ヘミン溶液
Ｈｅｍｉｎを１Ｎ・水酸化ナトリウム水溶液に０．１ｇ／２ｍＬで溶解し、精製水を加えて全量を２０ｍＬとし、オートクレーブ（１２１℃、１５分間）した。
（２）Ｖｉｔａｍｉｎ Ｋ１ ｓｔｏｃｋ ｓｏｌｕｔｉｏｎ
Ｖｉｔａｍｉｎ Ｋ１（０．９８４ｇ／ｍＬ）をエタノールに０．２ｍＬ／２０ｍＬで溶解し、ｖｉｔａｍｉｎ Ｋ１ ｓｔｏｃｋ ｓｏｌｕｔｉｏｎとした。
（３）メナジオン溶液
Ｖｉｔａｍｉｎ Ｋ１ ｓｔｏｃｋ ｓｏｌｕｔｉｏｎを滅菌精製水で１ｍＬ／９ｍＬに希釈したものをメナジオン溶液とした。
（４）５％ＬＨＢ添加Ｂｒｕｃｅｌｌａ ｂｒｏｔｈ
Ｂｒｕｃｅｌｌａ ｂｒｏｔｈを精製水に２８ｇ／Ｌで溶解し、１ｍＬのヘミン溶液及び１ｍＬのメナジオン溶液を加えてオートクレーブ（１２１℃、１５分間）した。適温に冷却後、馬溶血液を５０ｍＬ加えた。なお、使用する前には、４時間以上、嫌気条件下で予備還元した。
（５）ＧＡＭ ｂｒｏｔｈ
ＧＡＭブイヨンを精製水で５９ｇ／Ｌで溶解し、オートクレーブ（１２１℃、１５分間）した。なお、ＧＡＭ ｂｒｏｔｈは菌液調製に使用する前に、４時間以上、嫌気条件下で予備還元した。
４）試験薬液の調製
各試験物質を適量の０．１Ｎ・水酸化ナトリウム水溶液に溶解させた後、滅菌精製水で希釈して、４０×ＭＩＣの濃度の試験薬液の原液を調製した。
その後、フィルター濾過し、５％ＬＨＢ添加ｂｒｕｃｅｌｌａ ｂｒｏｔｈで１０倍希釈し、試験薬液を調製した。
５）試験菌液の調製
アネロコロンビアウサギ血液寒天培地を用いて２〜５日間、３５℃、嫌気条件下で前培養した菌株を滅菌生理食塩液２ｍＬに０．５ ＭｃＦａｒｌａｎｄ（約１．５×１０８ＣＦＵ／ｍＬ）となるように懸濁した後、１ｍＬをＧＡＭ ｂｒｏｔｈ ４ｍＬに加え、試験菌液（約３×１０７ＣＦＵ／ｍＬ）を調製した。その後、３時間、３５℃、嫌気条件下で培養した。
６）測定プレートの調製、菌の播種及び培養
試験薬液を１５ｍＬチューブに４．５０ｍＬずつ分注した。なお、薬物無処置群については、５％ＬＨＢ添加ｂｒｕｃｅｌｌａ ｂｒｏｔｈを用いた。
各チューブに試験菌液を５００μＬずつ添加した（初発菌濃度：約３×１０６ＣＦＵ／ｍＬ）。
２時間、３５℃、嫌気条件下で静置培養した。
菌懸濁液中に存在する試験物質を除去するため、各チューブをよく懸濁し、１．６ｍＬを２ｍＬチューブに分注し、遠心分離した（１００００ｒｐｍ、１分間、室温）。
上清を約１．５５ｍＬ取り、残余を１．５５ｍＬの生理食塩水で再懸濁し、遠心分離した（１００００ｒｐｍ、１分間、室温）。
上清を約１．５５ｍＬ取り、残余を１．５５ｍＬの５％ＬＨＢ添加ｂｒｕｃｅｌｌａ ｂｒｏｔｈで再懸濁し、遠心分離した（１００００ｒｐｍ、１分間、室温）。
上清を約１．５５ｍＬ取り、残余を１．５５ｍＬの５％ＬＨＢ添加ｂｒｕｃｅｌｌａ ｂｒｏｔｈで再懸濁した。
９６ウェルプレートに再懸濁液を２００μＬずつ添加した。
測定時間（試験物質除去後、０時間、１時間、１２時間、１５時間、１８時間、２１時間、２４時間）まで３５℃、嫌気条件下で静置培養した。
７）生菌数の定量
各ウェルから１００μＬ取り、滅菌生理食塩液９００μＬと混合し、１０１倍〜１０７倍の１０倍希釈系列を作製した。希釈した菌液２５μＬをアネロコロンビアウサギ血液寒天培地の半面に滴下し、コンラージ塗抹した。寒天培地は、３５℃で３〜５日間、嫌気培養後、発育したコロニー数を測定し生菌数を求めた。
生菌数のカウントは、Ｃｌｉｎｉｃａｌ Ｍｉｃｒｏｂｉｏｌｏｇｙ Ｐｒｏｃｅｄｕｒｅｓ Ｈａｎｄｂｏｏｋ ３ｒｄ ｅｄ．（ＡＳＭ、２０１０ ｕｐｄａｔｅ）に準じ、寒天培地でおおよそ３００コロニーとなる倍率と、それ以上の希釈倍率のコロニー数をカウントすることにより行った。
８）結果
評価した試験物質の中で、化合物Ａのみが、プロピオニバクテリウム・アクネスに対して明らかなＰＡＥを有しており、増殖抑制作用を有することが確認された。
[Test Example 1] PAE evaluation of various antibacterial drugs
1) Test substances In this test, the following antibacterial drugs were used as test substances.
Compound A (titer: 99.7%)
・ Nadifloxacin (Titer: 99.9%)
・ Levofloxacin (titer: 96.8%)
2) Used strain Propionibacterium acnes isolated from acne vulgaris patients in 2012-2013 was used.
The minimum inhibitory concentration (MIC) (μg / mL) of each test substance with respect to the strain used was as follows, and it was confirmed that the test substance was sensitive to each test substance.
In addition, the measurement of MIC was performed according to the micro liquid dilution method recommended by the Clinical and Laboratory Standards Institute.
Compound A (0.03)
・ Nadifloxacin (0.125)
・ Levofloxacin (0.5)
3) Preparation of medium (1) Hemin solution Hemin was dissolved in 1 N sodium hydroxide aqueous solution at 0.1 g / 2 mL, purified water was added to make a total volume of 20 mL, and autoclaved (121 ° C., 15 minutes).
(2) Vitamin K1 stock solution
Vitamin K1 (0.984 g / mL) was dissolved in ethanol at 0.2 mL / 20 mL to obtain vitamin K1 stock solution.
(3) Menadione solution Vitamin K1 stock solution diluted to 1 mL / 9 mL with sterilized purified water was used as the menadione solution.
(4) 5% LHB added Brucella broth
Brucella broth was dissolved in purified water at 28 g / L, 1 mL of hemin solution and 1 mL of menadione solution were added, and autoclaved (121 ° C., 15 minutes). After cooling to an appropriate temperature, 50 mL of equine hemolyzed blood was added. Prior to use, preliminary reduction was performed under anaerobic conditions for 4 hours or more.
(5) GAM broth
GAM bouillon was dissolved in purified water at 59 g / L and autoclaved (121 ° C., 15 minutes). GAM broth was pre-reduced under anaerobic conditions for 4 hours or more before being used for preparation of the bacterial solution.
4) Preparation of test drug solution Each test substance was dissolved in an appropriate amount of 0.1 N sodium hydroxide aqueous solution and then diluted with sterilized purified water to prepare a test drug solution stock solution having a concentration of 40 × MIC.
Thereafter, the mixture was filtered and diluted 10-fold with 5% LHB-added brucella broth to prepare a test drug solution.
5) Preparation of test bacterial solution The strain pre-cultured under anaerobic conditions at 35 ° C. for 2 to 5 days using Anero Columbia rabbit blood agar medium was added to 2 mL of sterile physiological saline solution with 0.5 McFarland (about 1.5 × 10 × 10). After suspension to 8 CFU / mL, 1 mL was added to 4 mL of GAM broth to prepare a test bacterial solution (about 3 × 10 7 CFU / mL). Then, it culture | cultivated on 35 degreeC and anaerobic conditions for 3 hours.
6) Preparation of measurement plate, seeding of fungus, and culture test chemical solution were dispensed into a 15 mL tube by 4.50 mL each. For the drug-untreated group, 5% LHB-added brucella broth was used.
500 μL of the test bacterial solution was added to each tube (initial bacterial concentration: about 3 × 10 6 CFU / mL).
Static culture was performed for 2 hours at 35 ° C. under anaerobic conditions.
In order to remove the test substance present in the bacterial suspension, each tube was well suspended, 1.6 mL was dispensed into a 2 mL tube, and centrifuged (10000 rpm, 1 minute, room temperature).
About 1.55 mL of the supernatant was taken, and the residue was resuspended in 1.55 mL of physiological saline and centrifuged (10000 rpm, 1 minute, room temperature).
About 1.55 mL of the supernatant was taken, and the residue was resuspended in 1.55 mL of 5% LHB-added brucella broth and centrifuged (10000 rpm, 1 minute, room temperature).
About 1.55 mL of the supernatant was taken, and the residue was resuspended in 1.55 mL of 5% LHB-added brucella broth.
200 μL of resuspension was added to each 96-well plate.
The cells were statically cultured under anaerobic conditions at 35 ° C. until the measurement time (0 hour, 1 hour, 12 hours, 15 hours, 18 hours, 21 hours, 24 hours after removal of the test substance).
7) Determination of the number of viable bacteria 100 μL was taken from each well and mixed with 900 μL of sterile physiological saline to prepare a 10-fold dilution series of 10 1 to 10 7 times. 25 μL of the diluted bacterial solution was dropped on one side of the Anero Columbia rabbit blood agar and smeared with Conage. The agar medium was subjected to anaerobic culture at 35 ° C. for 3 to 5 days, and the number of grown colonies was measured to determine the number of viable bacteria.
Viable counts were obtained from the Clinical Microbiology Procedures Handbook 3rd ed. According to (ASM, 2010 update), it was carried out by counting the number of colonies with a magnification of approximately 300 colonies on the agar medium and a dilution ratio of more than that.
8) Among the test substances evaluated as a result, it was confirmed that only Compound A has a clear PAE against Propionibacterium acnes and has a growth inhibitory action.
Claims (6)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2016089964 | 2016-04-28 | ||
| JP2016089964 | 2016-04-28 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2017200911A true JP2017200911A (en) | 2017-11-09 |
| JP7347915B2 JP7347915B2 (en) | 2023-09-20 |
Family
ID=60264287
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2017088146A Active JP7347915B2 (en) | 2016-04-28 | 2017-04-27 | antibacterial agent |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP7347915B2 (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999051588A1 (en) * | 1998-04-06 | 1999-10-14 | Toyama Chemical Co., Ltd. | Quinolonecarboxylic acid derivatives or salts thereof |
| JP2002356426A (en) * | 2001-03-26 | 2002-12-13 | Toyama Chem Co Ltd | External preparation for skin containing pyridonecarboxylic acid compound |
| WO2007015453A1 (en) * | 2005-08-01 | 2007-02-08 | Maruho Co., Ltd. | Lotion preparation containing pyridonecarboxylic acid derivative |
| JP2012505867A (en) * | 2008-10-17 | 2012-03-08 | フエルレル インターナショナル，ソシエダッド アノニマ | Topical pharmaceutical composition |
-
2017
- 2017-04-27 JP JP2017088146A patent/JP7347915B2/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999051588A1 (en) * | 1998-04-06 | 1999-10-14 | Toyama Chemical Co., Ltd. | Quinolonecarboxylic acid derivatives or salts thereof |
| JP2002356426A (en) * | 2001-03-26 | 2002-12-13 | Toyama Chem Co Ltd | External preparation for skin containing pyridonecarboxylic acid compound |
| WO2007015453A1 (en) * | 2005-08-01 | 2007-02-08 | Maruho Co., Ltd. | Lotion preparation containing pyridonecarboxylic acid derivative |
| JP2012505867A (en) * | 2008-10-17 | 2012-03-08 | フエルレル インターナショナル，ソシエダッド アノニマ | Topical pharmaceutical composition |
Non-Patent Citations (3)
| Title |
|---|
| JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY, vol. 49, JPN6020050628, 2002, pages 455 - 465, ISSN: 0004531862 * |
| JOURNAL OF MEDICAL MICROBIOLOGY, vol. 65, JPN6020050626, August 2016 (2016-08-01), pages 745 - 750, ISSN: 0004691282 * |
| 臨床医薬, vol. 31, JPN6020050623, 2015, pages 155 - 171, ISSN: 0004691281 * |
Also Published As
| Publication number | Publication date |
|---|---|
| JP7347915B2 (en) | 2023-09-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11529361B2 (en) | Halogenated salicylanilides for treating Clostridium infections | |
| WAISBREN et al. | A clinical appraisal of neomycin | |
| EP2826473B1 (en) | Antibacterial use of patchoulol | |
| JP2008535814A (en) | Antimicrobial agent | |
| Nahar et al. | Association of biofilm formation with antimicrobial resistance among the Acinetobacter species in a tertiary care hospital in Bangladesh | |
| CN103565798B (en) | The novelty teabag of Pogostone and derivant thereof | |
| BRPI1010948B1 (en) | antibacterial composition, pharmaceutical composition or medication, uses of the composition and in vitro process for preparing a surface | |
| CN106620649A (en) | Methods of treatment using single doses of oritavancin | |
| Yendewa et al. | A two-part phase 1 study to establish and compare the safety and local tolerability of two nasal formulations of XF-73 for decolonisation of Staphylococcus aureus: A previously investigated 0.5 mg/g viscosified gel formulation versus a modified formulation | |
| JPWO2010147145A1 (en) | Anti-gram negative bacteria agent | |
| US20240316068A1 (en) | Methods for treating hypervirulent klebsiella pneumoniae infection | |
| CN110652512A (en) | Application of crizotinib in preparation of anti-gram-positive-bacteria drugs | |
| CN119345170A (en) | A composition and its application in fighting multidrug-resistant Acinetobacter baumannii | |
| RU2640928C2 (en) | Combination of fulvic acid and antibiotic for suppression of growth of bacteria resistant to majority of drugs, or treatment of infections caused by them | |
| JP2017200911A (en) | Antimicrobial agent | |
| WO2002022147A1 (en) | Antibacterial combination comprising neem plant extract | |
| CN115531395A (en) | Application of deoxycholic acid in preparation of product for resisting streptococcus infantis | |
| Farca et al. | In vitro antimicrobial activity of marbofloxacin and enrofloxacin against bacterial strains isolated from companion animals | |
| CN115350197A (en) | Application of alisol A-24-acetate in improving sensitivity of MRSA to beta-lactam antibiotics | |
| Hernandez-Romero et al. | Rifampicin and N-acteylcisteyne inhibit oral bacterial growth and biofilm formation | |
| Al-Sabbagh et al. | Comparison between the Effect of Normal and Nanoparticles of ZnO Against Different Types of Bacteria Isolated from Patients with Acne | |
| KR102208837B1 (en) | Composition for inhibiting adhesion, invasion of bacteria or antibacterial resistance comprising methyl gallate and fluoroquinolone antibacterial agent | |
| AU2018285342B2 (en) | Antibiotic composition for the treatment of infections with resistent microorganism | |
| Scherr et al. | Buruli ulcer | |
| Copenhagen | ATX201 REDUCES STAPHYLOCOCCUS AUREUS COLONIZATION AND INCREASES SHANNON DIVERSITY OF THE SKIN MICROBIOME IN ATOPIC DERMATITIS PATIENTS |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| RD01 | Notification of change of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7426 Effective date: 20170511 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20170511 |
|
| RD02 | Notification of acceptance of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7422 Effective date: 20180405 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20180405 |
|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20200207 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20200219 |
|
| RD02 | Notification of acceptance of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7422 Effective date: 20200219 |
|
| A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20201221 |
|
| RD02 | Notification of acceptance of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7422 Effective date: 20201223 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20201224 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20210203 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20210303 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20210623 |
|
| A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20220126 |
|
| C60 | Trial request (containing other claim documents, opposition documents) |
Free format text: JAPANESE INTERMEDIATE CODE: C60 Effective date: 20220405 |
|
| C22 | Notice of designation (change) of administrative judge |
Free format text: JAPANESE INTERMEDIATE CODE: C22 Effective date: 20230419 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20230707 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20230907 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 7347915 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |