JP2004331632A - 2-fluoro-6-o-substituted ketoride derivative - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new erythromycin derivative having a potent anti-bacterial activity against haemophilus influenza and erythromycin resistant pneumococcus, and having less drug interactions. <P>SOLUTION: This new erythromycin derivative is a 2-fluoro-6-O-substituted ketoride derivative expressed by formula (1) or its medicinally acceptable salt. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

【０００８】
で表される２−フルオロ−６−Ｏ−置換ケトライド誘導体又はその医薬上許容される塩である。
【０００９】
【発明の実施形態】
本発明において、医薬上許容される塩とは、細菌感染症の化学療法および予防において使用される塩を意味する。それらは、たとえば酢酸、プロピオン酸、酪酸、ギ酸、トリフルオロ酢酸、マレイン酸、酒石酸、クエン酸、ステアリン酸、コハク酸、エチルコハク酸、ラクトビオン酸、グルコン酸、グルコヘプトン酸、安息香酸、メタンスルホン酸、エタンスルホン酸、２−ヒドロキシエタンスルホン酸、ベンゼンスルホン酸、パラトルエンスルホン酸、ラウリル硫酸、リンゴ酸、アスパラギン酸、グルタミン酸、アジピン酸、システイン、Ｎ−アセチルシステイン、塩酸、臭化水素酸、リン酸、硫酸、ヨウ化水素酸、ニコチン酸、シュウ酸、ピクリン酸、チオシアン酸、ウンデカン酸、アクリル酸ポリマー、カルボキシビニルポリマーなどの酸との塩を挙げることができる。
【００１０】
本発明の２−フルオロ−６−Ｏ−置換ケトライド誘導体は、経口投与又は非経口投与され、成人を治療する場合で５０〜１０００ｍｇであり、これを１日２〜３回に分けて投与する。この投与量は、患者の年齢、体重および症状によって適宜増減することができる。
【００１１】
経口投与する場合は、賦形剤、結合剤、滑沢剤、抗酸化剤、コーティング剤、界面活性剤、可塑剤、着色剤、矯味矯臭剤などを混合して、散剤、顆粒剤、カプセル剤、錠剤などの製剤として投与され、非経口投与する場合は、注射剤、点滴剤などの製剤として投与される。製剤化する際には、通常の製剤化の方法が使用できる。
【００１２】
【発明の効果】
本発明の化合物は、エリスロマイシン耐性肺炎球菌及びインフルエンザ菌に対して優れた抗菌活性を有し、感染動物を用いた治療効果にも優れていること、更にはヒト代謝酵素に対しても安定な事から、本発明の化合物はヒトにおけるエリスロマイシン耐性肺炎球菌及びインフルエンザ菌感染症の治療剤として極めて有用である。
【００１３】
【実施例】
次に、参考例、実施例及び試験例にて本発明を更に詳細に説明する。
【００１４】
参考例１
５−ヨード−３−（ピリミジン−２−イル）イソキサゾールの合成（２−シアノピリミジンを用いる方法）
（１）２−シアノピリミジン３ｇ（２８．５ｍｍｏｌ）、メタノール ５０ｍｌ、ヒドロキシルアミン塩酸塩１．９８ｇ（２８．５ｍｍｏｌ）、ナトリウムメトキシド１．５４ｇ（２８．５ｍｍｏｌ）を順次加え、室温で２時間攪拌した。反応混合物を減圧濃縮の後残渣に水５０ｍｌを加え沈殿物を濾取、得られた粗生成物をエーテルで洗浄した後、減圧乾燥し、２−（Ｎ−ヒドロキシアミジノ）ピリミジン ２．８ｇ（収率７１％）を得た。
【００１５】
ＭＳ（ＥＳＩ） ｍ／ｚ １６１．０［Ｍ＋Ｎａ］^＋
１Ｈ−ＮＭＲ（２００ ＭＨｚ， ＤＭＳＯ−ｄ_６） δ（ｐｐｍ）：５．８３ （ｓ， ２ Ｈ），７．５２ （ｔ， Ｊ＝４．８３ Ｈｚ， １ Ｈ），８．８５ （ｄ， Ｊ＝４．８３ Ｈｚ， ２ Ｈ），１０．１８ （ｓ， １ Ｈ）
【００１６】
（２）２−（Ｎ−ヒドロキシアミジノ）ピリミジン２．７ｇ（１９．５ｍｍｏｌ）を水５４ｍｌに懸濁し、氷冷下濃塩酸２０ｍｌを徐々に加えた。さらに亜硝酸ナトリウム１．７ｇ（２４．２ｍｍｏｌ）を水９．３ｍｌに溶かした溶液を１０分かけて滴下した。その後氷冷下で２時間攪拌した後、反応混合物を炭酸水素ナトリウムにより中和した。析出した結晶を濾取し、減圧乾燥して２−（クロロ−Ｎ−ヒドロキシイミドイル）ピリミジン２．２８ｇ（収率７４％）を得た。
【００１７】
ＭＳ（ＥＳＩ） ｍ／ｚ １７９．９［Ｍ＋Ｎａ］^＋
１Ｈ−ＮＭＲ（２００ ＭＨｚ， ＤＭＳＯ−ｄ_６） δ（ｐｐｍ）：７．６１ （ｔ， Ｊ＝５．０５ Ｈｚ， １ Ｈ），８．９３ （ｄ， Ｊ＝４．８３ Ｈｚ， ２ Ｈ），１２．９４ （ｓ， １ Ｈ）
元素分析値（Ｃ_５Ｈ_４ＣｌＮ_３Ｏ） 理論値： Ｃ，３８．１２；Ｈ，２．５６；Ｎ，２６．６７；Ｃｌ，２２．５０，測定値： Ｃ，３８．０１；Ｈ，２．５３；Ｎ，２６．７２；Ｃｌ，２２．３２
【００１８】
（３）エチニルトリ（ｎ−ブチル）スズ２５ｇ（７９．３ｍｍｏｌ）をＴＨＦ５０ｍｌに溶解し氷冷下ヨウ素２１．８ｇ（８６ｍｍｏｌ）を少しずつ加えた。このまま２０分攪拌した後、２−（クロロ−Ｎ−ヒドロキシイミドイル）ピリミジン２．０ｇ（１３．２ｍｍｏｌ）を一度に加え、続いてトリエチルアミン４．１ｍｌ（２９ｍｍｏｌ）をＴＨＦ１２ｍｌに溶かした溶液を５分かけて滴下した。反応混合物を室温で９０分攪拌し、これを６％チオ硫酸ナトリウム水溶液と酢酸エチルの混合物にあけ分液した。有機層を硫酸マグネシウムで乾燥後減圧濃縮し、残渣をシリカゲルカラムクロマトグラフィー（ヘキサン−酢酸エチル＝３／１〜酢酸エチル）により精製して標題化合物１．７１ｇ（収率４９％）を得た。
【００１９】
ＭＳ（ＥＳＩ） ｍ／ｚ ２９５．８［Ｍ＋Ｎａ］^＋
１Ｈ−ＮＭＲ（２００ ＭＨｚ， ＣＤＣｌ_３） δ（ｐｐｍ）：７．１８ （ｓ， １ Ｈ），７．３９ （ｔ， Ｊ＝４．８３ Ｈｚ， １ Ｈ），８．９０ （ｄ， Ｊ＝４．８３ Ｈｚ， ２ Ｈ）
【００２０】
参考例２
５−ヨード−３−（ピリミジン−２−イル）イソキサゾールの合成（２−メチルピリミジンを用いる方法）
（１）カリウムｔ−ブトキシド８．９ｇ（８０ｍｏｌ）をＴＨＦ ５０ｍｌに溶解し氷冷下２−メチルピリミジン５ｇ（５３ｍｍｏｌ）をＴＨＦ１５ｍｌに溶かした溶液を約５分かけて滴下した。反応混合物を室温で１．５時間攪拌した後、氷冷下亜硝酸ｔ−ブチル １２．６ｍｌ（１０６ｍｍｏｌ）をＴＨＦ１０ｍｌに溶かした溶液を約５分かけて滴下し、その後室温で１９時間攪拌した。反応混合物を減圧濃縮し水２０ｍｌを加え、その後４Ｎ−塩酸水溶液を加え中和した。この混合物を減圧濃縮し、析出した結晶を濾取して２−ピリミジンアルドキシムの高極性異性体 ２．７５ｇを得た。母液は塩析して酢酸エチルで抽出（５０ｍｌ×２）、有機層を硫酸マグネシウムで乾燥後減圧濃縮した。残渣にヘキサンを加え、得られた沈殿物を濾取して、２−ピリミジンアルドキシムの異性体混合物（低極性異性体：高極性異性体＝７：２）２．２３ｇを得た。合計４．９８ｇ（収率７６％）のオキシム体を得た。
【００２１】
低極性異性体
ＭＳ（ＥＳＩ） ｍ／ｚ １４５．９［Ｍ＋Ｎａ］^＋
１Ｈ−ＮＭＲ（２００ ＭＨｚ， ＤＭＳＯ−ｄ_６） ｄ（ｐｐｍ）：７．５９ （ｔ， Ｊ＝５．０５ Ｈｚ， １ Ｈ），７．７８ （ｓ， １ Ｈ），８．９６ （ｄ， Ｊ＝４．８３ Ｈｚ， ２ Ｈ），１３．４５ （ｓ， １ Ｈ）
高極性異性体
ＭＳ（ＥＳＩ） ｍ／ｚ １４５．９［Ｍ＋Ｎａ］^＋
１Ｈ−ＮＭＲ（２００ ＭＨｚ， ＤＭＳＯ−ｄ_６） δ（ｐｐｍ）：７．４７ （ｔ， Ｊ＝５．０５ Ｈｚ， １ Ｈ），８．１１ （ｓ， １ Ｈ），８．８４ （ｄ， Ｊ＝４．８３ Ｈｚ， ２ Ｈ），１２．０１ （ｓ， １ Ｈ）
【００２２】
（２）３００ｍｌナスフラスコに、上記２−ピリミジンアルドキシム異性体混合物４．９ｇ（４０ｍｍｏｌ）、酢酸エチル１００ｍｌ、トリブチルエチニルスズ１０．４ｇ（３３ｍｍｏｌ）、炭酸水素ナトリウム８．４ｇ（１００ｍｍｏｌ）、水５ｍｌ、Ｎ−クロロコハク酸イミド５．３ｇ（４０ｍｍｏｌ）を順次加え、室温で１６時間攪拌した。水５ｍｌをさらに加え４時間攪拌した後、飽和炭酸水素ナトリウム水溶液１００ｍｌを加え分液した。水層を酢酸エチルで抽出後有機層を合わせ、飽和食塩水で洗浄、硫酸マグネシウムで乾燥した。減圧濃縮の後ヘキサンを加え析出した沈殿物を濾別し、母液を減圧濃縮してトリ−ｎ−ブチル−｛［３−（ピリミジン−２−イル）］イソキサゾール−５−イル｝スズの粗生成物１３．６ｇを得た。
【００２３】
この粗生成物１３．６ｇをＴＨＦ１００ｍｌに溶解し、ヨウ素７．９ｇ（３１．２ｍｍｏｌ）を一度に加えた。反応混合物を室温で３５分攪拌した後反応混合物を６％チオ硫酸ナトリウム水溶液にあけ、酢酸エチルで抽出した。有機層を飽和食塩水で洗浄、硫酸マグネシウムで乾燥後減圧濃縮し、残渣にヘキサンを加え析出している結晶を濾取した。この結晶をヘキサンで洗浄、乾燥して標題化合物 ５．４４ｇ（収率６０％）を得た。
【００２４】
ＭＳ（ＥＳＩ） ｍ／ｚ ２９５．９［Ｍ＋Ｎａ］^＋
１Ｈ−ＮＭＲ（３００ ＭＨｚ， ＣＤＣｌ_３） δ（ｐｐｍ）：７．１８ （ｓ， １ Ｈ），７．３８ （ｔ， Ｊ＝４．９０ Ｈｚ， １ Ｈ），８．８９ （ｄ， Ｊ＝４．８２ Ｈｚ， ２ Ｈ）
【００２５】
実施例１
５−Ｏ−デソサミニル−３，１１−ジデオキシ−２−フルオロ−１１−アミノ−６−Ｏ−［３−［３−（２−ピリミジル）イソキサゾリル−５−イル］−２−プロピニル］−３−オキソエリスロノリドＡ １１，１２−サイクリックカーバメートの合成
ＷＯ９９２１８７１および米国特許第６１２４２６９号に記した方法により合成した５−Ｏ−デソサミニル−３，１１−ジデオキシ−２−フルオロ−１１−アミノ−３−オキソ−６−Ｏ−プロパルギルエリスロノリドＡ １１，１２−サイクリックカーバメート５．５ｇ （８．４ｍｍｏｌ）、５−ヨード−３−（２−ピリミジル）イソキサゾール２．４ｇ （８．８ｍｍｏｌ）、ビス（トリフェニルホスフィン）パラジウムクロリド（ＩＩ） ０．２９ｇ （０．４ｍｍｏｌ）、アセトニトリル４５ｍｌ、トリエチルアミン９ｍｌを混合し、系内をアルゴン置換した後室温で１時間、６０度で１４時間攪拌した。反応混合物を減圧濃縮して得られた残渣を、シリカゲルカラムクロマトグラフィー（クロロホルム：メタノール：２５％アンモニア水＝１９：１：０．１）、次いでＮＨ型シリカゲルカラムクロマトグラフィー（酢酸エチル）により精製し、イソプロピルエーテルにて固化、濾取、乾燥して標記化合物 ５．６ｇ（収率８３％）を得た。
ＮＭＲ（３００ＭＨｚ， ＣＤＣｌ_３） δ（ｐｐｍ）：０．９３ （ｔ， Ｊ＝７．４６ Ｈｚ， ３ Ｈ），１．１３−１．１９ （ｍ， ３ Ｈ），１．１８ （ｄ， Ｊ＝７．０１ Ｈｚ， ３ Ｈ），１．２０−１．７０ （ｍ， ２ Ｈ），１．２５ （ｄ， Ｊ＝６．０９ Ｈｚ， ３ Ｈ），１．３４ （ｄ， Ｊ＝７．３１ Ｈｚ， ３ Ｈ），１．５２ （ｓ， ３ Ｈ），１．５６ （ｓ， ３ Ｈ），１．６２−２．０５ （ｍ， ２ Ｈ），１．６８−１．８８ （ｍ， ２ Ｈ），１．８１ （ｄ， Ｊ＝２１．３２ Ｈｚ， ３ Ｈ），２．２７ （ｓ， ６ Ｈ），２．４６ （ｄｄｄ， Ｊ＝１２．２６， １０．２８， ３．６５ Ｈｚ， １ Ｈ），２．６９ （ｍ， １ Ｈ），２．９４ （ｍ， １ Ｈ），３．１９ （ｄｄ， Ｊ＝１０．０５， ７．３１ Ｈｚ， １ Ｈ），３．５５ （ｍ， １ Ｈ），３．６６ （ｍ， １ Ｈ），３．７１ （ｓ， １ Ｈ），３．８４ （ｄｄ， Ｊ＝４４．７７， １７．３６ Ｈｚ， ２ Ｈ），４．１７ （ｄｄ， Ｊ＝１０．２０， １．３７ Ｈｚ， １ Ｈ），４．３４ （ｄ， Ｊ＝７．３１ Ｈｚ， １ Ｈ），５．０７ （ｄｄ， Ｊ＝１０．０５， ２．７４ Ｈｚ， １ Ｈ），５．６５ （ｓ， １ Ｈ），７．３２ （ｓ， １ Ｈ），７．３５ （ｔ， Ｊ＝４．８７ Ｈｚ， １ Ｈ），８．８９ （ｄ， Ｊ＝４．８７ Ｈｚ， ２ Ｈ）
【００２６】
試験例１：試験管内抗菌活性測定
被験化合物は、実施例１の化合物と比較化合物１としてＷＯ０２／３２９１９の実施例２３記載の化合物（５−Ｏ−デソサミニル−３，１１−ジデオキシ−２−フルオロ−１１−アミノ−６−Ｏ−［３−［３−（５−ピリミジル）イソキサゾリル−５−イル］−２−プロピニル］−３−オキソエリスロノリドＡ １１，１２−サイクリックカーバメート）及び比較化合物２としてアジスロマイシンを選びエリスロマイシン耐性肺炎球菌及びインフルエンザ菌に対する抗菌活性を、Ｎａｔｉｏｎａｌ Ｃｏｍｍｉｔｔｅｅ ｆｏｒ Ｃｌｉｎｉｃａｌ Ｌａｂｏｒａｔｏｒｙ Ｓｔａｎｄａｒｄｓ （ＮＣＣＬＳ） ｇｕｉｄｅｌｉｎｅｓに準じた微量液体希釈法による最小発育阻止濃度（ＭＩＣ）で評価した。各々羊血液寒天培地およびチョコレート寒天培地で一夜培養した肺炎球菌及びインフルエンザ菌をＭｕｅｌｌｅｒ−Ｈｉｎｔｏｎ ｂｒｏｔｈ （ＭＨＢ）で０．５ ＭｃＦａｒａｎｄ相当に懸濁し、これらを１０倍に希釈（約１０^７ ＣＦＵ／ｍｌ）して接種用菌液とした。これらの菌液５ ｍｌを３％馬溶血液添加の２価イオン濃度調整済みＭＨＢおよびＨａｅｍｏｐｈｉｌｕｓ ｔｅｓｔ ｍｅｄｉｕｍに約５×１０^５ ＣＦＵ／ｍｌの菌量となるように感受性試験用マイクロプレートに接種した。これらを３５℃、通常環境下で２０時間培養後に各化合物のＭＩＣを測定した。これら試験の結果を表１に示す。実施例１の化合物はエリスロマイシン耐性肺炎球菌に対し比較化合物と同様強い抗菌活性を示した。インフルエンザ菌に対しては比較化合物２の１／２であって比較化合物１の２倍の抗菌活性を示した。本願発明の化合物は、エリスロマイシン耐性肺炎球菌及びインフルエンザ菌の両方に有用であることが示された。
【００２７】
【表１】

【００２８】
試験例２：インフルエンザ菌による肺炎に対する治療効果
インフルエンザ菌による肺炎に対する治療効果について、インフルエンザ菌によるマウス肺炎モデルで評価した。被験化合物及びインフルエンザ菌は、試験例１と同様である。インフルエンザ菌を、５％馬脱繊維血液添加Ｂｒａｉｎ Ｈｅａｒｔ Ｉｎｆｕｓｉｏｎ Ｂｒｏｔｈで３７℃一夜培養し、これを接種菌液（約５．０×１０^８ ＣＦＵ／ｍｌ）とした。麻酔した体重１８〜２２ ｇのＩＣＲ系雄性マウスに接種菌液０．０５ ｍｌを経気道的に接種することにより、感染を惹起した。菌液接種１８および４２時間後に経口ゾンデを用いて各被験化合物１００ｍｇ／ｋｇを経口投与した（各群５匹）。各被験化合物は５％アラビアゴム溶液に用時懸濁し、希釈した。なお、コントロール群には５％アラビアゴム溶液のみを投与した。最終投与２４時間後にマウスを屠殺し、肺を摘出後、２ ｍｌの滅菌生理食塩液を加えてホモジナイズした。滅菌生理食塩液で１０倍希釈系列を作製し、チョコレート寒天培地に塗布して３７℃一夜培養後、生じたコロニー数から生菌数を算出した。なお、検出限界は２．３０ ｌｏｇ_１０ ＣＦＵ／ｍｌである。これら試験の結果を表２に示す。被験化合物１００ ｍｇ／ｋｇの投与群において、本願発明の化合物である実施例１の化合物、比較化合物１及び比較化合物２はコントロールに比べいずれも強い抗菌活性を示した。よって、本願発明の化合物は、比較化合物１及び比較化合物２と同様にインフルエンザ菌による肺炎の治療に有用であることが示された。
【００２９】
【表２】

【００３０】
試験例３
エリスロマイシン耐性肺炎球菌による肺炎に対する治療効果について、耐性肺炎球菌によるマウス肺炎モデルで評価した。被験化合物としては、実施例１の化合物及び比較化合物１としてＷＯ０２／３２９１９の実施例２３記載の化合物（５−Ｏ−デソサミニル−３，１１−ジデオキシ−２−フルオロ−１１−アミノ−６−Ｏ−［３−［３−（５−ピリミジル）イソキサゾリル−５−イル］−２−プロピニル］−３−オキソエリスロノリドＡ １１，１２−サイクリックカーバメート）を用いた。試験例１で用いたエリスロマイシン耐性肺炎球菌を３０％馬血清添加Ｔｏｄｄ−Ｈｅｗｉｔｔ Ｂｒｏｔｈで３７℃数時間培養後、同培地で３００倍に希釈し、接種菌液（約５×１０^５ ＣＦＵ／ｍｌ）とした。麻酔した体重１８〜２２ ｇのＣＢＡ／Ｊ系雄性マウスに接種菌液０．０５ ｍｌを経鼻的に接種することにより、感染を惹起した。菌液接種４２および６８時間後に経口ゾンデを用いて各被験化合物１００ ｍｇ／ｋｇを経口投与した（各群５匹）。各被験化合物は５％アラビアゴム溶液に用時懸濁し、希釈した。なお、コントロール群には５％アラビアゴム溶液のみを投与した。最終投与２４時間後にマウスを屠殺し、肺を摘出後、２ ｍｌの滅菌生理食塩液を加えてホモジナイズした。滅菌生理食塩液で１０倍希釈系列を作製し、羊血液寒天培地に塗布して３７℃、５％炭酸ガス存在下で一夜培養後、生じたコロニー数から生菌数を算出した。なお、検出限界は１．３０ ｌｏｇ_１０ ＣＦＵ／ｌｕｎｇである。これら試験の結果を表３に示す。
本願発明の化合物である実施例１の化合物は、１００ｍｇ／ｋｇ投与群において比較化合物と同程度の優れた治療効果を示した。
【００３１】
【表３】

【００３２】
試験例４：ヒト型Ｃｙｐ阻害試験
Ｃｙｐ阻害試験は被験化合物を添加した後、蛍光基質を添加する前に４５分間のプレインキュベーションを行った。その他はＣｒｅｓｐｉらの方法（Ｃｒｅｓｐｉ ＣＬ， ｅｔ ａｌ．， Ａｎａｌｙｔ． Ｂｉｏｃｈｅｍ． １９９７， ２４８， １８８−１９０）に準拠して行った。
【００３３】
被験化合物としては、試験例３と同様である。コントロールとして、クラリスロマイシンを用いた。
【００３４】
【表４】

【００３５】
実施例１の化合物の相互作用の強さはコントロールと比べほぼ同程度であったが、比較化合物の阻害作用は実施例１の化合物の約８６倍と非常に強かった。よって、比較化合物１に比べて、実施例１の化合物はＣｙｐ３Ａ４に対するＩＣ_５０値が高く、薬物相互作用を起こす可能性が低いことが示された。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to novel derivatives of the antibiotic erythromycin.
[0002]
[Prior art]
Erythromycin A is an antibiotic widely used as a therapeutic drug for infectious diseases caused by gram-positive bacteria, mycoplasma and the like. However, since erythromycin is decomposed by stomach acid, there is a drawback that pharmacokinetics are not constant. Therefore, derivatives having increased acid stability have been studied, and as a result, macrolide agents having stable pharmacokinetics, such as clarithromycin, azithromycin and roxithromycin, have been developed. These macrolides which are used for treatment of exogenous respiratory tract infections need to have a strong antibacterial activity against influenza bacteria and pneumococci, which are frequently clinically isolated. Furthermore, since macrolide-resistant pneumococci are frequently isolated from community-acquired pneumonia, it is also important to be effective against resistant pneumococci.
[0003]
In recent years, extensive studies have shown that Agouridas et al. In 1995 found HMR3647 (Terithromycin, EP680967) as a macrolide effective against both Influenza and erythromycin resistant pneumococci, and then Or et al. In 1998 ABT-773. (Seslomycin, WO9809978). Thereafter, 2-fluoroketolide (WO 02/32919), which has further enhanced the efficacy, has been reported. On the other hand, 14-membered macrolide antibiotics are known to cause drug interactions. Therefore, it is of course very important that the macrolide agent not only exerts a sufficient therapeutic effect in the clinic but also does not easily cause drug interaction. The drug interaction caused by the 14-membered macrolide antibiotic is caused by the fact that a molecular species involved in metabolism (Cyp3A4) metabolizes the 14-membered macrolide and the metabolized macrolide binds irreversibly to Cyp3A4. Has been caused. Therefore, it is important to be stable to human Cyp3A4.
[0004]
[Problems to be solved by the invention]
An object of the present invention is to provide a novel 2-fluoro-6-O-substituted ketolide having excellent antibacterial activity against Haemophilus influenzae and erythromycin highly resistant pneumococcus, and being stable against human Cyp3A4 metabolism. .
[0005]
[Means for Solving the Problems]
The present inventors have conducted various studies on ketolide derivatives, and as a result, a ketolide introduced with a propargyl group having a specific heterocyclic group at the 6-position and further with a fluorine atom introduced at the 2-position has excellent antibacterial activity, Then, they found that they were stable against human Cyp3A4, and completed the present invention.
[0006]
The present invention provides a compound of the formula
Embedded image

[0008]
Is a 2-fluoro-6-O-substituted ketolide derivative or a pharmaceutically acceptable salt thereof.
[0009]
DETAILED DESCRIPTION OF THE INVENTION
In the present invention, the pharmaceutically acceptable salt means a salt used in chemotherapy and prevention of bacterial infection. They include, for example, acetic, propionic, butyric, formic, trifluoroacetic, maleic, tartaric, citric, stearic, succinic, ethylsuccinic, lactobionic, gluconic, glucoheptonic, benzoic, methanesulfonic, Ethanesulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, paratoluenesulfonic acid, lauryl sulfate, malic acid, aspartic acid, glutamic acid, adipic acid, cysteine, N-acetylcysteine, hydrochloric acid, hydrobromic acid, phosphoric acid And salts with acids such as sulfuric acid, hydroiodic acid, nicotinic acid, oxalic acid, picric acid, thiocyanic acid, undecanoic acid, acrylic acid polymer and carboxyvinyl polymer.
[0010]
The 2-fluoro-6-O-substituted ketolide derivative of the present invention is orally administered or parenterally administered, and is 50 to 1000 mg when treating an adult, and is administered twice or three times a day. This dosage can be appropriately increased or decreased depending on the age, weight and condition of the patient.
[0011]
For oral administration, excipients, binders, lubricants, antioxidants, coating agents, surfactants, plasticizers, coloring agents, flavoring agents, etc. are mixed, and powders, granules, capsules In the case of parenteral administration, it is administered as a preparation such as an injection or a drip. In formulating, a usual method of formulating can be used.
[0012]
【The invention's effect】
The compounds of the present invention have excellent antibacterial activity against erythromycin-resistant pneumococci and Haemophilus influenzae, have excellent therapeutic effects using infected animals, and are also stable against human metabolic enzymes. Therefore, the compounds of the present invention are extremely useful as therapeutic agents for erythromycin-resistant pneumococcal and H. influenzae infections in humans.
[0013]
【Example】
Next, the present invention will be described in more detail with reference examples, examples and test examples.
[0014]
Reference Example 1
Synthesis of 5-iodo-3- (pyrimidin-2-yl) isoxazole (method using 2-cyanopyrimidine)
(1) 3 g (28.5 mmol) of 2-cyanopyrimidine, 50 ml of methanol, 1.98 g (28.5 mmol) of hydroxylamine hydrochloride and 1.54 g (28.5 mmol) of sodium methoxide are sequentially added, and the mixture is stirred at room temperature for 2 hours. did. After the reaction mixture was concentrated under reduced pressure, 50 ml of water was added to the residue, and the precipitate was collected by filtration. The obtained crude product was washed with ether, dried under reduced pressure, and 2.8 g of 2- (N-hydroxyamidino) pyrimidine (yield) Rate 71%).
[0015]
MS (ESI) m / z 161.0 [M + Na] ^+
1 H-NMR (200 MHz, DMSO-d ₆ ) δ (ppm): 5.83 (s, 2 H), 7.52 (t, J = 4.83 Hz, 1 H), 8.85 (d , J = 4.83 Hz, 2 H), 10.18 (s, 1 H)
[0016]
(2) 2.7 g (19.5 mmol) of 2- (N-hydroxyamidino) pyrimidine was suspended in 54 ml of water, and 20 ml of concentrated hydrochloric acid was gradually added under ice-cooling. Further, a solution of 1.7 g (24.2 mmol) of sodium nitrite dissolved in 9.3 ml of water was added dropwise over 10 minutes. After stirring for 2 hours under ice cooling, the reaction mixture was neutralized with sodium hydrogen carbonate. The precipitated crystals were collected by filtration and dried under reduced pressure to obtain 2.28 g (yield 74%) of 2- (chloro-N-hydroxyimidoyl) pyrimidine.
[0017]
MS (ESI) m / z 179.9 [M + Na] ^< +>
¹ H-NMR (200 MHz, DMSO-d ₆ ) δ (ppm): 7.61 (t, J = 5.05 Hz, 1 H), 8.93 (d, J = 4.83 Hz, 2 H) ), 12.94 (s, 1 H)
Elemental analysis _{(C 5 H 4 ClN 3 O} ) theory: C, 38.12; H, 2.56 ; N, 26.67; Cl, 22.50, measurements: C, 38.01; H, 2.53; N, 26.72; Cl, 22.32.
[0018]
(3) 25 g (79.3 mmol) of ethynyltri (n-butyl) tin was dissolved in 50 ml of THF, and 21.8 g (86 mmol) of iodine was added little by little under ice-cooling. After stirring for 20 minutes as it is, 2.0 g (13.2 mmol) of 2- (chloro-N-hydroxyimidoyl) pyrimidine was added at once, and then a solution of 4.1 ml (29 mmol) of triethylamine dissolved in 12 ml of THF was added for 5 minutes. The solution was dropped. The reaction mixture was stirred at room temperature for 90 minutes, poured into a mixture of 6% sodium thiosulfate aqueous solution and ethyl acetate, and separated. The organic layer was dried over magnesium sulfate and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (hexane-ethyl acetate = 3 / 1-ethyl acetate) to obtain 1.71 g (yield 49%) of the title compound.
[0019]
MS (ESI) m / z 295.8 [M + Na] ^+
1 H-NMR (200 MHz, CDCl ₃ ) δ (ppm): 7.18 (s, 1 H), 7.39 (t, J = 4.83 Hz, 1 H), 8.90 (d, J = 4.83 Hz, 2 H)
[0020]
Reference Example 2
Synthesis of 5-iodo-3- (pyrimidin-2-yl) isoxazole (method using 2-methylpyrimidine)
(1) A solution prepared by dissolving 8.9 g (80 mol) of potassium t-butoxide in 50 ml of THF and dissolving 5 g (53 mmol) of 2-methylpyrimidine in 15 ml of THF was added dropwise under ice-cooling over about 5 minutes. After the reaction mixture was stirred at room temperature for 1.5 hours, a solution of 12.6 ml (106 mmol) of t-butyl nitrite dissolved in 10 ml of THF was added dropwise over about 5 minutes under ice cooling, and then stirred at room temperature for 19 hours. The reaction mixture was concentrated under reduced pressure, 20 ml of water was added, and then a 4N aqueous hydrochloric acid solution was added for neutralization. The mixture was concentrated under reduced pressure, and the precipitated crystals were collected by filtration to obtain 2.75 g of a highly polar isomer of 2-pyrimidinealdoxime. The mother liquor was salted out and extracted with ethyl acetate (50 ml × 2). The organic layer was dried over magnesium sulfate and concentrated under reduced pressure. Hexane was added to the residue, and the obtained precipitate was collected by filtration to obtain 2.23 g of a mixture of isomers of 2-pyrimidinealdoxime (low-polar isomer: high-polar isomer = 7: 2). 4.98 g (yield 76%) of the oxime was obtained in total.
[0021]
Less polar isomer MS (ESI) m / z 145.9 [M + Na] ^+
1 H-NMR (200 MHz, DMSO-d ₆ ) d (ppm): 7.59 (t, J = 5.05 Hz, 1 H), 7.78 (s, 1 H), 8.96 (d , J = 4.83 Hz, 2 H), 13.45 (s, 1 H)
Highly polar isomer MS (ESI) m / z 145.9 [M + Na] ^+
1 H-NMR (200 MHz, DMSO-d ₆ ) δ (ppm): 7.47 (t, J = 5.05 Hz, 1 H), 8.11 (s, 1 H), 8.84 (d , J = 4.83 Hz, 2 H), 12.01 (s, 1 H)
[0022]
(2) In a 300 ml eggplant flask, 4.9 g (40 mmol) of the above 2-pyrimidine aldoxime isomer mixture, 100 ml of ethyl acetate, 10.4 g (33 mmol) of tributylethynyltin, 8.4 g (100 mmol) of sodium hydrogen carbonate, and 5 ml of water And 5.3 g (40 mmol) of N-chlorosuccinimide were sequentially added, followed by stirring at room temperature for 16 hours. After 5 ml of water was further added and the mixture was stirred for 4 hours, 100 ml of a saturated aqueous solution of sodium hydrogencarbonate was added and the mixture was separated. After the aqueous layer was extracted with ethyl acetate, the organic layers were combined, washed with brine, and dried over magnesium sulfate. After concentration under reduced pressure, hexane was added, and the deposited precipitate was separated by filtration. The mother liquor was concentrated under reduced pressure to give crude tri-n-butyl-{[3- (pyrimidin-2-yl)] isoxazol-5-yl} tin. 13.6 g of the product were obtained.
[0023]
13.6 g of this crude product was dissolved in 100 ml of THF, and 7.9 g (31.2 mmol) of iodine was added at a time. After the reaction mixture was stirred at room temperature for 35 minutes, the reaction mixture was poured into a 6% aqueous sodium thiosulfate solution and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. Hexane was added to the residue, and the precipitated crystals were collected by filtration. The crystals were washed with hexane and dried to give the title compound (5.44 g, yield 60%).
[0024]
MS (ESI) m / z 295.9 [M + Na] ^< +>
¹ H-NMR (300 MHz, CDCl ₃ ) δ (ppm): 7.18 (s, 1 H), 7.38 (t, J = 4.90 Hz, 1 H), 8.89 (d, J = 4.82 Hz, 2 H)
[0025]
Example 1
5-O-desosaminyl-3,11-dideoxy-2-fluoro-11-amino-6-O- [3- [3- (2-pyrimidyl) isoxazolyl-5-yl] -2-propynyl] -3-oxo Synthesis of erythronolide A 11,12-cyclic carbamate 5-O-desosaminyl-3,11-dideoxy-2-fluoro-11-amino-3-oxo synthesized by the method described in WO9921871 and U.S. Pat. No. 6,124,269. 5.5 g (8.4 mmol) of -6-O-propargylerythronolide A 11,12-cyclic carbamate, 2.4 g (8.8 mmol) of 5-iodo-3- (2-pyrimidyl) isoxazole, bis (tri Phenylphosphine) palladium chloride (II) 0.29 g (0.4 mmol), acetonitrile 4 ml, triethylamine 9ml mixed, in the system for one hour at room temperature after the atmosphere was replaced with argon and stirred at 60 ° for 14 hours. The residue obtained by concentrating the reaction mixture under reduced pressure was purified by silica gel column chromatography (chloroform: methanol: 25% aqueous ammonia = 19: 1: 0.1) and then NH-type silica gel column chromatography (ethyl acetate). Then, the residue was solidified with isopropyl ether, collected by filtration and dried to obtain 5.6 g (yield 83%) of the title compound.
NMR (300 MHz, CDCl ₃ ) δ (ppm): 0.93 (t, J = 7.46 Hz, 3 H), 1.13-1.19 (m, 3 H), 1.18 (d, J = 7.01 Hz, 3H), 1.20-1.70 (m, 2H), 1.25 (d, J = 6.09 Hz, 3H), 1.34 (d, J = 7 .31 Hz, 3 H), 1.52 (s, 3 H), 1.56 (s, 3 H), 1.62-2.05 (m, 2 H), 1.68-1.88 ( m, 2H), 1.81 (d, J = 21.32 Hz, 3H), 2.27 (s, 6H), 2.46 (ddd, J = 12.26, 10.28, 3 .65 Hz, 1 H), 2.69 (m, 1 H), 2.94 (m, 1 H), 3.19 (dd, J = 10.05, 7.31 Hz, 1 H), 3 .55 ( , 1H), 3.66 (m, 1H), 3.71 (s, 1H), 3.84 (dd, J = 44.77, 17.36 Hz, 2H), 4.17 ( dd, J = 10.20, 1.37 Hz, 1H), 4.34 (d, J = 7.31 Hz, 1H), 5.07 (dd, J = 10.05, 2.74 Hz) , 1H), 5.65 (s, 1H), 7.32 (s, 1H), 7.35 (t, J = 4.87 Hz, 1H), 8.89 (d, J = 4.87 Hz, 2 H)
[0026]
Test Example 1: In Vitro Antibacterial Activity Measurement The test compounds were the compound of Example 1 and the compound (5-O-desosaminyl-3,11-dideoxy-2-fluoro-) described in Example 23 of WO 02/32919 as Comparative Compound 1. 11-amino-6-O- [3- [3- (5-pyrimidyl) isoxazolyl-5-yl] -2-propynyl] -3-oxoerythronolide A 11,12-cyclic carbamate) and comparative compound 2 Azithromycin was selected as the product, and the antibacterial activity against erythromycin-resistant pneumococci and Haemophilus influenzae was evaluated by a minimum growth inhibitory concentration (MIC) by a microfluidic dilution method according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines. Streptococcus pneumoniae and Haemophilus influenzae cultured overnight on sheep blood agar medium and chocolate agar medium, respectively, were suspended in Mueller-Hinton broth (MHB) to an equivalent of 0.5 McFarland, and diluted 10-fold (about 10 ⁷ CFU / ml). This was used as a bacterial solution for inoculation. 5 ml of these bacterial solutions were inoculated into a susceptibility test microplate so as to give a bacterial amount of about 5 × 10 ⁵ CFU / ml in MHB and Haemophilus test medium in which the divalent ion concentration was adjusted with the addition of 3% horse lysate. After culturing them at 35 ° C. in a normal environment for 20 hours, the MIC of each compound was measured. Table 1 shows the results of these tests. The compound of Example 1 showed strong antibacterial activity against erythromycin-resistant pneumococcus like the comparative compound. The antibacterial activity against H. influenzae was 1/2 that of Comparative Compound 2 and twice as high as that of Comparative Compound 1. The compounds of the present invention have been shown to be useful for both erythromycin resistant pneumococci and Haemophilus influenzae.
[0027]
[Table 1]

[0028]
Test Example 2: Therapeutic effect on pneumonia caused by Haemophilus influenzae The therapeutic effect on pneumonia caused by Haemophilus influenzae was evaluated using a mouse pneumonia model caused by Haemophilus influenzae. The test compound and H. influenzae were the same as in Test Example 1. The Haemophilus influenzae was cultured overnight at 37 ° C. in Brain Heart Infusion Broth supplemented with 5% horse defibrinated blood, and this was used as an inoculum (about 5.0 × 10 ⁸ CFU / ml). Infection was induced by intraperitoneally inoculating 0.05 ml of the inoculated bacterial solution into anesthetized ICR male mice weighing 18 to 22 g. Eighteen and 42 hours after inoculation of the bacterial solution, 100 mg / kg of each test compound was orally administered using an oral probe (5 mice per group). Each test compound was suspended and diluted in a 5% gum arabic solution before use. The control group received only a 5% gum arabic solution. Twenty-four hours after the final administration, the mice were sacrificed, and the lungs were excised and homogenized by adding 2 ml of sterile physiological saline. A 10-fold dilution series was prepared with sterile physiological saline, applied to a chocolate agar medium, cultured at 37 ° C. overnight, and the number of viable bacteria was calculated from the number of colonies formed. The detection limit is 2.30 log ₁₀ CFU / ml. Table 2 shows the results of these tests. In the 100 mg / kg test compound administration group, the compound of Example 1 of the present invention, Comparative Compound 1 and Comparative Compound 2 all showed stronger antibacterial activity than the control. Therefore, the compound of the present invention was shown to be useful for treating pneumonia caused by Haemophilus influenzae, similarly to Comparative Compound 1 and Comparative Compound 2.
[0029]
[Table 2]

[0030]
Test example 3
The therapeutic effect on pneumonia caused by erythromycin resistant pneumococci was evaluated in a mouse pneumonia model caused by resistant pneumococci. As test compounds, the compound of Example 1 and the compound described in Example 23 of WO 02/32919 (5-O-desosaminyl-3,11-dideoxy-2-fluoro-11-amino-6-O-O- [3- [3- (5-Pyrimidyl) isoxazolyl-5-yl] -2-propynyl] -3-oxoerythronolide A 11,12-cyclic carbamate) was used. After culturing the erythromycin-resistant pneumococcus used in Test Example 1 in Todd-Hewitt Broth supplemented with 30% horse serum for several hours at 37 ° C., it was diluted 300-fold with the same medium, and the inoculated bacterial solution (about 5 × 10 ⁵ CFU / ml) And Infection was induced by intranasally inoculating an anesthetized CBA / J male mouse weighing 18 to 22 g with 0.05 ml of the inoculum. At 42 and 68 hours after inoculation of the bacterial solution, 100 mg / kg of each test compound was orally administered using an oral probe (5 mice per group). Each test compound was suspended and diluted in a 5% gum arabic solution before use. The control group received only a 5% gum arabic solution. Twenty-four hours after the final administration, the mice were sacrificed, and the lungs were excised and homogenized by adding 2 ml of sterile physiological saline. A 10-fold dilution series was prepared with sterile physiological saline, applied to sheep blood agar medium, cultured overnight at 37 ° C. in the presence of 5% carbon dioxide, and the number of viable bacteria was calculated from the number of colonies formed. The detection limit is 1.30 log ₁₀ CFU / lung. Table 3 shows the results of these tests.
The compound of Example 1, which is the compound of the present invention, showed an excellent therapeutic effect comparable to that of the comparative compound in the 100 mg / kg administration group.
[0031]
[Table 3]

[0032]
Test Example 4: Human Cyp Inhibition Test In the Cyp inhibition test, pre-incubation was performed for 45 minutes after adding the test compound and before adding the fluorescent substrate. Others were performed according to the method of Crespi et al. (Crespi CL, et al., Analyt. Biochem. 1997, 248, 188-190).
[0033]
The test compound is the same as in Test Example 3. Clarithromycin was used as a control.
[0034]
[Table 4]

[0035]
The interaction strength of the compound of Example 1 was almost the same as that of the control, but the inhibitory effect of the comparative compound was very strong, about 86 times that of the compound of Example 1. Therefore, compared to Comparative Compound 1, the compound of Example 1 had a higher IC ₅₀ value for Cyp3A4, indicating a lower possibility of causing drug interaction.

Claims (1)

formula

A 2-fluoro-6-O-substituted ketolide derivative represented by the formula: or a pharmaceutically acceptable salt thereof.