JPH0250108B2

JPH0250108B2 -

Info

Publication number: JPH0250108B2
Application number: JP63205389A
Authority: JP
Inventors: Tsutomu Irikura; Hiroshi Koga; Satoru Murayama
Original assignee: Kyorin Pharmaceutical Co Ltd
Current assignee: Kyorin Pharmaceutical Co Ltd
Priority date: 1988-08-18
Filing date: 1988-08-18
Publication date: 1990-11-01
Also published as: JPH01258666A

Description

【発明の詳細な説明】[Detailed description of the invention]

本発明は新規置換キノリンカルボン酸に関す
る。ナリジクス酸、ピロミド酸及びピペミド酸はグ
ラム陰性菌感染症に有効な安全性の高い合成抗菌
剤として臨床的に広く用いられている。しかしな
がら、これらの薬剤は、グラム陽性菌あるいは難
治性感染症の原因菌の一つであり有効な薬剤が少
ない緑膿菌に対して非常に弱い抗菌力しか有して
いない。本発明者らは、先に１−（２−フルオロ
エチル）−１，４−ジヒドロ−４−オキソキノリ
ン−３−カルボン酸の６位にフツ素原子、７位に
ピペラジン又は置換ピペラジンを有する化合物が
緑膿菌を含むグラム陰性菌並びにグラム陽性菌に
対して強い抗菌力を有することを報告した［日本
薬学会第98年会（昭和53年４月、岡山）、講演要
旨集、P.233、特開昭55−47658号］。本発明者ら
はその後この関連化合物について、更に検討し、
その内から１位にフルオロエチル基、６位及び８
位にフツ素原子、７位に４−メチルピペラジンを
同時に有する６，７，８−トリ置換体は、６，７
−又は７，８−ジ置換体よりもより強い抗菌力を
有し、またこの化合物は試験管内では構造的に近
縁の化合物と略同等であるが、より実際面の用途
に近い生体内の試験では高い抗菌力が見出され
た。すなわち、本発明は下記の式［］で表される
置換キノリンカルボン酸並びにその生理学的に許容される塩及び水和物を
提供するものである。また、式［］の化合物の塩としてはナトリウ
ム、カリウム、カルシウム、マグネシウム、アル
ミニウム、セリウム、クロム、コバルト、銅、
鉄、銀、亜鉛等の金属塩及び有機塩基の塩、更に
塩酸、硫酸、リン酸等の無機酸塩、酢酸、乳酸、
メタンスルホン酸等の有機酸塩等が例示できる。本発明の化合物は例えば下記製造法により得る
ことができる。すなわち式［］で表される化合物と次の式［］で表される化合物とを反応させることにより式
［］で表される置換キノリンカルボン酸、並びにその
塩及び水和物を製造できる。本反応は、溶媒中又は無溶媒下、脱酸剤の存在
下反応させるのが望ましいが、を過剰に用いて脱酸剤あるいは溶媒を兼ねさせて
もよい。次に本発明化合物は精薬組成物にも使用するこ
とができる。本発明化合物の人及び動物（魚類を含む）の治
療及び予防のためには一般の薬学的な投与形式が
用いられる。明確な投与形式及び投与基準は治療
又は予防する個体の状況に応じて適宜選択する事
ができる。剤型の代表的なものとして錠剤、丸剤、散剤、
液剤、懸濁剤、乳剤、顆粒剤、カプセル剤、坐
剤、注射剤、軟膏剤等を例示する事ができる。軟膏剤等の有機基剤を用いる製剤においては本
発明化合物中の銀、亜鉛、セリウム等の金属塩の
使用も好ましく、有機基剤との親和性にも優れて
いる。本発明の内容を更に具体的に説明するため、以
下の製造例を述べる。製造例６，７，８−トリフルオロ−１，４−ジヒドロ
−４−オキソキノリン−３−カルボン酸エチルエ
ステル0.8ｇ、無水炭酸カリウム2.2ｇ、１−ブロ
ム−２−フルオロエタン3.8ｇ及びヨウ化ナトリ
ウム4.5ｇをDMF30mlに加え90〜100℃で10時間
撹拌した。次いで溶媒を留去し残渣に水を加えて塩化メチ
レンで抽出し、水洗、乾燥の後溶媒を留去した。この残渣に18％塩酸14ml及びエタノール14ml及
び水14mlを加え氷冷し、析出結晶を濾取し、
DMF−エタノール混合溶媒（１：２）より再結
晶し、１−（２−フルオロエチル）−６，７，８−
トリフルオロ−１，４−ジヒドロ−４−オキソキ
ノリン−３−カルボン酸を白色粉末として0.34ｇ
を得た。 mp：208〜210℃ 分析値 C₁₂H₇NO₃F₄ ＣＨＮ計算値（％） 49.84 2.44 4.84 実測値（％） 49.86 2.37 5.01 上記化合物の１−（２−フルオロエチル）−６，
７，８−トリフルオロ−１，４−ジヒドロ−４−
オキソキノリン−３−カルボン酸0.12ｇ及び１−
メチルピペラジン0.34ｇをピリジン３mlに加え油
浴上140℃で６時間還流した。次いで溶媒を留去し酢酸でPHを４にし、熱時不
溶物を濾去した。この濾液に塩酸を加えPHを１に
し、析出結晶を濾取し、水から再結晶した。６，８−ジフルオロ−１−（２−フルオロエチ
ル）−１，４−ジヒドロ−４−オキソ−７−（４−
メチル−１−ピペラジニル）−キノリン−３−カ
ルボン酸塩酸塩を黄色針状晶として0.08ｇ得た。 mp：269〜271℃（分解）分析値 C₁₇H₁₈N₃C₃F₃・HCl ＣＨＮ計算値（％） 50.32 4.72 10.36 実測値（％） 50.12 4.97 10.24 参考例１上記製造例で得た本発明化合物について抗菌試
験を行つた。その結果は第１表に示す通りであ
る。試験方法は日本化学療法学会指定の方法［日
本化学療法学会編：最少発育阻止濃度（MIC）
測定法改定について、日本化学療法学会雑誌、22
巻６号、1126〜1128頁（1974年）］に準じた。本発明化合物は第１表に示したようにグラム陰
性菌に対し極めて強い抗菌活性を示すと同時にグ
ラム陽性菌の中でもブドウ球菌に対し強い活性を
示した。 The present invention relates to novel substituted quinoline carboxylic acids. Nalidixic acid, pyromidic acid, and pipemidic acid are widely used clinically as highly safe synthetic antibacterial agents effective against Gram-negative bacterial infections. However, these drugs have very weak antibacterial activity against Gram-positive bacteria or Pseudomonas aeruginosa, which is one of the causative bacteria of refractory infections and for which there are few effective drugs. The present inventors previously developed a compound having a fluorine atom at the 6-position and a piperazine or substituted piperazine at the 7-position of 1-(2-fluoroethyl)-1,4-dihydro-4-oxoquinoline-3-carboxylic acid. reported that it has strong antibacterial activity against Gram-negative bacteria including Pseudomonas aeruginosa as well as Gram-positive bacteria [98th Annual Meeting of the Pharmaceutical Society of Japan (April 1978, Okayama), Collection of Lecture Abstracts, p. 233 , Japanese Patent Publication No. 55-47658]. The present inventors then further investigated this related compound,
Fluoroethyl group at the 1st position, 6th and 8th positions
A 6,7,8-tri-substituted product having a fluorine atom at the 7-position and a 4-methylpiperazine at the 7-position is a 6,7-trisubstituted
- or 7,8-disubstituted compounds, and although this compound is structurally similar in vitro to closely related compounds, it has been shown to have stronger antibacterial activity than the 7,8-disubstituted derivatives, and although this compound is structurally similar in vitro, it has been tested in vivo, which is closer to practical applications. Tests showed high antibacterial activity. That is, the present invention provides a substituted quinoline carboxylic acid represented by the following formula [] and physiologically acceptable salts and hydrates thereof. In addition, salts of the compound of formula [] include sodium, potassium, calcium, magnesium, aluminum, cerium, chromium, cobalt, copper,
Metal salts such as iron, silver, zinc and salts of organic bases, as well as inorganic acid salts such as hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, lactic acid,
Examples include organic acid salts such as methanesulfonic acid. The compound of the present invention can be obtained, for example, by the following production method. That is, the expression [] The compound represented by and the following formula [] By reacting with a compound represented by the formula [] Substituted quinoline carboxylic acids represented by the following formula, as well as salts and hydrates thereof, can be produced. This reaction is preferably carried out in a solvent or in the absence of a solvent, and in the presence of a deoxidizing agent. may be used in excess to serve as a deoxidizing agent or a solvent. Next, the compounds of the present invention can also be used in seminal drug compositions. Common pharmaceutical modes of administration are used for the treatment and prophylaxis of humans and animals (including fish) with the compounds of this invention. The precise administration format and administration standards can be appropriately selected depending on the circumstances of the individual to be treated or prevented. Typical dosage forms include tablets, pills, powders,
Examples include solutions, suspensions, emulsions, granules, capsules, suppositories, injections, and ointments. In preparations using organic bases such as ointments, it is also preferable to use metal salts such as silver, zinc, and cerium in the compounds of the present invention, which have excellent affinity with organic bases. In order to more specifically explain the content of the present invention, the following manufacturing examples will be described. Production example 6,7,8-trifluoro-1,4-dihydro-4-oxoquinoline-3-carboxylic acid ethyl ester 0.8 g, anhydrous potassium carbonate 2.2 g, 1-bromo-2-fluoroethane 3.8 g and iodide 4.5 g of sodium was added to 30 ml of DMF and stirred at 90-100°C for 10 hours. Next, the solvent was distilled off, water was added to the residue, extracted with methylene chloride, washed with water, dried, and the solvent was distilled off. To this residue were added 14 ml of 18% hydrochloric acid, 14 ml of ethanol, and 14 ml of water, cooled on ice, and the precipitated crystals were collected by filtration.
Recrystallized from DMF-ethanol mixed solvent (1:2), 1-(2-fluoroethyl)-6,7,8-
0.34g of trifluoro-1,4-dihydro-4-oxoquinoline-3-carboxylic acid as white powder
I got it. mp: 208-210℃ Analysis value C ₁₂ H ₇ NO ₃ F ₄ C H N Calculated value (%) 49.84 2.44 4.84 Actual value (%) 49.86 2.37 5.01 1-(2-fluoroethyl)-6 of the above compound,
7,8-trifluoro-1,4-dihydro-4-
Oxoquinoline-3-carboxylic acid 0.12g and 1-
0.34 g of methylpiperazine was added to 3 ml of pyridine, and the mixture was refluxed on an oil bath at 140°C for 6 hours. Then, the solvent was distilled off, the pH was adjusted to 4 with acetic acid, and insoluble materials were filtered off when heated. Hydrochloric acid was added to this filtrate to adjust the pH to 1, and the precipitated crystals were collected by filtration and recrystallized from water. 6,8-difluoro-1-(2-fluoroethyl)-1,4-dihydro-4-oxo-7-(4-
0.08 g of methyl-1-piperazinyl)-quinoline-3-carboxylic hydrochloride was obtained as yellow needles. mp: 269-271℃ (decomposition) Analytical value C ₁₇ H ₁₈ N ₃ C ₃ F ₃・HCl C H N Calculated value (%) 50.32 4.72 10.36 Actual value (%) 50.12 4.97 10.24 Reference example 1 Obtained in the above production example Antibacterial tests were conducted on the compounds of the present invention. The results are shown in Table 1. The test method is the method specified by the Japanese Society of Chemotherapy [edited by the Japanese Society of Chemotherapy: Minimum Inhibitory Concentration (MIC)]
Regarding the revision of the measurement method, Journal of the Japanese Society of Chemotherapy, 22
Vol. 6, pp. 1126-1128 (1974)]. As shown in Table 1, the compounds of the present invention exhibited extremely strong antibacterial activity against Gram-negative bacteria, and also exhibited strong activity against Staphylococcus among Gram-positive bacteria.

【表】参考例２特開昭55−47658号の実施例22，23，30，31を
比較化合物１〜４とし、これら及びノルフロキサ
シンについて、上記参考例１と同様の試験を行な
い、その結果を本発明化合物と比較して第２表に
示した。 [Table] Reference Example 2 Examples 22, 23, 30, and 31 of JP-A-55-47658 were used as Comparative Compounds 1 to 4, and the same tests as in Reference Example 1 above were conducted on these and norfloxacin. A comparison with the compounds of the present invention is shown in Table 2.

【表】【table】

【表】上記第２表の試験管内抗菌活性の試験結果か
ら、本発明化合物は比較化合物と同等の広い抗菌
活性を有する有用性が認められた。またこれに加え、本発明化合物は優れた生体内
抗菌活性が認められ、比較化合物に比べて下記の
如くその生体内抗菌活性は特に優れていた。参考例３マウス全身感染治療実験による生体内抗菌活
性、及び比較化合物２，３，４との比較本発明化合物の生体内における抗菌活性をマウ
ス全身感染治療実験で調べ、その結果を下記第３
−１表に示した。また比較化合物２，３，４と比較してその結果
を第３−２表に示した。全身感染マウスは雄性マウス（体重20±２ｇ）
の腹腔内に、５％ムチンを含む滅菌したブレンハ
ートインフユージヨンブイヨン0.5mlに病原菌を
懸濁（大腸菌のみは滅菌生理食塩水0.5mlに懸濁）
したものを注射して作製した。化合物は感染直後と４時間後の２回経口投与
し、その後１週間観察して、動物の50％を救うの
に有効な投与量（ED₅₀）をプロビツト法により
計算し判定した。本発明化合物は各菌種による全身感染防御効果
において比較化合物２，３，４に比べ著しく優れ
ていた。なお比較化合物１は、大腸菌（E.coli
ML4707）に対する生体内抗菌活性において下記
第３−１表に示す通り殆ど効果が無かつたので他
の試験は行なわなかつた。[Table] From the in vitro antibacterial activity test results shown in Table 2 above, the compounds of the present invention were found to have the same broad antibacterial activity as the comparative compounds. In addition, the compound of the present invention was found to have excellent in-vivo antibacterial activity, and its in-vivo antibacterial activity was particularly superior to that of comparative compounds as described below. Reference Example 3 In vivo antibacterial activity in a mouse systemic infection treatment experiment and comparison with comparative compounds 2, 3, and 4 The in vivo antibacterial activity of the compound of the present invention was investigated in a mouse systemic infection treatment experiment, and the results were summarized in Section 3 below.
-1 Shown in Table. The results were also compared with Comparative Compounds 2, 3, and 4, and the results are shown in Table 3-2. Systemically infected mice are male mice (body weight 20±2g)
Intraperitoneally, suspend pathogenic bacteria in 0.5 ml of sterilized Blenhart infusion broth containing 5% mucin (suspend only Escherichia coli in 0.5 ml of sterile physiological saline).
It was made by injecting it. The compound was orally administered twice, immediately after infection and 4 hours later, and the animals were then observed for one week, and the dose effective for saving 50% of the animals (ED ₅₀ ) was calculated and determined by the probit method. The compounds of the present invention were significantly superior to Comparative Compounds 2, 3, and 4 in terms of protective effects against systemic infections caused by various bacterial species. Comparative Compound 1 was prepared using Escherichia coli (E.coli).
As shown in Table 3-1 below, there was almost no effect on in vivo antibacterial activity against ML4707), so no other tests were conducted.

【表】【table】

【表】参考例４チヤイニーズハムスターCHL細胞に対する細
胞毒性についての本発明化合物と比較化合物
２，３，４との比較本発明化合物の動物細胞に対する毒性をチヤイ
ニーズハムスターCHL細胞の生育阻害実験で調
べ、比較化合物２，３，４と比較し、結果は第４
表に示した。 10％牛胎仔血清を含むイーグルスミニマムエツ
センシヤル培養液に細胞を３×10⁴cells／mlの濃
度に懸濁し、その懸濁液（１ml）を上記培養液３
mlを含むガラスプレートを加え、37℃、５％炭酸
ガス−95％空気条件下で培養した。翌日、培養液
を50μｇ／mlの化合物を含む上記培養液４mlに交
換し、更に３日間培養した。３日後に生細胞数を
変え、各化合物について生育阻害パーセントを算
出した。本発明化合物は細菌細胞に対して前記第２表で
示す如く比較化合物２，３，４と概ね同等の試験
管内抗菌活性を有するが、動物細胞に対する毒性
は比較化合物２，３，４に比して更に低く、選択
毒性に優れている。[Table] Reference Example 4 Comparison of the compound of the present invention and Comparative Compounds 2, 3, and 4 regarding cytotoxicity to Chinese hamster CHL cells Growth inhibition experiment of Chinese hamster CHL cells to test the toxicity of the compound of the present invention to animal cells and compared with comparative compounds 2, 3, and 4, and the results are as follows:
Shown in the table. Cells were suspended in Eagles Minimum Essential Culture Medium containing 10% fetal bovine serum at a concentration of 3 x 10 ⁴ cells/ml, and the suspension (1 ml) was added to the above culture medium 3.
ml was added to a glass plate and cultured at 37°C under 5% carbon dioxide-95% air conditions. The next day, the culture medium was replaced with 4 ml of the above culture medium containing 50 μg/ml of the compound, and the cells were cultured for an additional 3 days. After 3 days, the number of viable cells was changed and the percent growth inhibition was calculated for each compound. The compound of the present invention has approximately the same in vitro antibacterial activity against bacterial cells as Comparative Compounds 2, 3, and 4 as shown in Table 2 above, but its toxicity against animal cells is lower than that of Comparative Compounds 2, 3, and 4. It has even lower toxicity and excellent selective toxicity.

【表】【table】

Claims

【特許請求の範囲】１式［］で表される置換キノリンカルボン酸並びにその生
理学的に許容される塩及び水和物。[Claims] 1 Formula [] Substituted quinoline carboxylic acids represented by: and physiologically acceptable salts and hydrates thereof.