JP2004502736A - Macrolide formulations for inhalation and methods of treating endobronchial infections - Google Patents

Abstract

Described are macrolide formulations (eg, erythromycylamine formulations) for delivery by aerosol application. The concentrated erythromycylamine formulation contains an effective amount of erythromycylamine to treat infections caused by susceptible bacteria. A unit dose device having a container containing the formulation of the macrolide antibiotic in a physiologically acceptable carrier is also described. Also described are methods for the treatment of pulmonary infections by formulations (liquid solutions, suspensions, or dry powders) delivered as aerosols having a median aerodynamic diameter of primarily between 1-5 μm. You.

Description

この処方物のｐＨは、エアロゾル送達に同等に重要である。以前に注記したように、エアロゾルが酸性または塩基性のいずれかである場合、エアロゾルは気管支痙攣および咳を生じ得る。ｐＨの安全性範囲は相対的であり；幾人かの患者は、他の患者では気管支痙攣を生じる温和な酸性のエアロゾルを耐容する。４．５未満のｐＨを有する任意のエアロゾルは、通常過敏な個体では気管支痙攣を誘導する；４．５と５．０との間のｐＨを有するエアロゾルは、この問題をたまに生じる。５．０と７．０との間のｐＨを有するエアロゾルは、安全であると考えられる。１０．０より大きいｐＨを有する任意のエアロゾルは、回避されるべきである。なぜなら気管支痙攣を生じる刺激が生じ得るからである。エアロゾル処方物に対する至適ｐＨは、ｐＨ５．０と７．０との間であることが決定された。
【００４５】
１つの局面において、本発明の液体処方物は、好ましくは主に、細菌が存在する末端細気管支および呼吸細気管支、ならびに気道下部への、この薬物の送達を可能にする粒子サイズに噴霧される。エアロゾルによる肺気管支内気道へのエリスロマイシルアミンの効率的な送達のためには、主に１〜５μｍの間の大きさのメジアン空気力学的直径を有するエアロゾル粒子の処方物が必要である。気管支内感染（特に細菌であるＳｔｒｅｐｔｏｃｏｃｃｕｓ　ｐｎｅｕｍｏｎｉａｅ、Ｈａｅｍｏｐｈｉｌｕｓ　ｉｎｆｌｕｅｎｚａｅ、Ｓｔａｐｈｙｌｏｃｃｏｕｓ　ａｕｒｅｕｓ、Ｍｏｒａｘｅｌｌａ　ｃａｔａｒｒｈａｌｉｓ、Ｌｅｇｉｏｎｅｌｌａ　ｐｎｅｕｍｏｎｉａ、Ｃｈｌａｍｙｄｉａ　ｐｎｅｕｍｏｎｉａｅ、およびＭｙｃｏｐｌａｓｍａ　ｐｎｅｕｍｏｎｉａｅによって起こる感染）の処置および予防のためのエリスロマイシルアミンの処方された量および送達された量は、気管支内表面を効率的に標的しなければならない。処方物の送達された用量は、好ましくは、感染の部位へ有効な用量のエリスロマイシルアミンを送達できるのに最小である粒子エアロゾール化可能な容積を有する。好ましい処方物は、気道の機能に有害に影響しない条件をさらに提供する。結果として、好ましい処方物は、望ましくない反応は回避しながら、薬物の効率的な送達を可能にする条件下で処方されたその薬物の十分な量を含む。本発明による新規な処方物は、これらの要求の全てを満たす。
【００４６】
本発明に従って、エリスロマイシルアミンは、慢性気管支炎および肺炎を有する患者による吸入療法のために意図された投薬形態に処方される。患者は世界中に存在するので、この処方物は合理的に長期の保存期間を有することが所望される。従って、保管条件および処方物の安定性は重要となる。
【００４７】
上記で考察したように、この溶液のｐＨは重要である。保管およびより長期間の保存期間の観点から、５．０と７．０の間のｐＨ、好ましくは約６．０のｐＨが至適である。
【００４８】
この処方物は代表的に、１〜２ミリリットルの低密度ポリエチレン（ＬＤＰＥ）バイアル中に保管される。このバイアルは、吹き込み充填シール（ｂｌｏｗ−ｆｉｌｌ−ｓｅａｌ）プロセスを用いて無菌的に充填される。このバイアルは、ホイルオーバーパウチ（ｆｏｉｌ　ｏｖｅｒｐｏｕｃｈ）で密閉（シール）される。
【００４９】
酸化に関するこの処方物の安定性は、別の非常に重要な問題である。この薬物がエアロゾル化の前に分解される場合、肺に送達される薬物はさらに少量になり、従って、送達された用量が非常に少なくなるので、処置を障害し、そしてエリスロマイシルアミンに対する耐性の発現を導き得る条件を誘発する。さらに、エリスロマイシルアミン分解産物は、気管支痙攣および咳を誘発し得る。エリスロマイシルアミンの酸化的分解を防止するため、そして受容可能な安定性を提供するため、ホイルオーバーパウチ（１オーバーパウチあたり６バイアル）を含む酸素保護包装（パッケージ）中にこのＬＤＰＥバイアルをパッケージングすることによって、低酸素含量の製品を生産する。バイアル充填の前に、混合タンク中の溶液を窒素噴霧し、そして環状のオーバーパウチの頭部空隙を窒素パージする。この方法で、エリスロマイシルアミンの加水分解および酸化の両方を防ぐ。
【００５０】
（ＩＩ．エアロゾル化デバイス）
本発明の実施に有用なエアロゾル化デバイス（例えば、ジェット噴霧器、振動多孔性（ビブレーティングポーラス）プレート噴霧器、または超音波噴霧器）は一般に、本発明の処方物を、主に１〜５μｍの範囲のエアロゾル粒子に噴霧できる。主にとは、本出願では、全ての生成されたエアロゾル粒子の少なくとも７０％、ただし好ましくは９０％より多くが１〜５μｍの範囲内であることを意味する。
【００５１】
噴霧器（例えば、ジェット噴霧器、超音波噴霧器、振動多孔性（ビブレーティングポーラス）プレート噴霧器）、および高電圧乾燥粉末吸入器（Ｓｔｒｅｐｔｏｃｏｃｃｕｓ　ｐｎｅｕｍｏｎｉａｅ、Ｈａｅｍｏｐｈｉｌｕｓ　ｉｎｆｌｕｅｎｚａｅ、Ｓｔａｐｈｙｌｏｃｃｏｕｓ　ａｕｒｅｕｓ、Ｍｏｒａｘｅｌｌａ　ｃａｔａｒｒｈａｌｉｓ、Ｌｅｇｉｏｎｅｌｌａ　ｐｎｅｕｍｏｎｉａ、Ｃｈｌａｍｙｄｉａ　ｐｎｅｕｍｏｎｉａｅ、およびＭｙｃｏｐｌａｓｍａ　ｐｎｅｕｍｏｎｉａｅの感染の処置に至適である、粒子サイズ１と５μｍとの間の粒子を生成し送達し得る）は、現在利用可能であるか、または公知の方法および材料を用いて作製され得る。ジェット噴霧器は、空気圧によって働き、液体溶液をエアロゾル小滴に破壊する。振動多孔性（ビブレーティングポーラス）プレート噴霧器は、素早く振動する多孔性プレートによって生成される音波減圧を用いることにより働いて、多孔性プレートを通って溶媒小滴を押し出す。超音波噴霧器は、液体を小さいエアロゾル小滴に分割する圧電性結晶によって働く。しかし、エリスロマイシルアミンのいくつかの処方物しか、これらの３つの噴霧器によって効率的に噴霧され得ない。なぜならこれらのデバイスは処方物のｐＨおよびイオン強度に感受性であるからである。
【００５２】
種々のデバイスが利用可能であるが、これらの噴霧器の限られた数しか本発明の目的に適切でない。本発明において有用な好ましい噴霧器としては、例えば、ＡｅｒｏＮｅｂ^ＴＭおよびＡｅｒｏＤｏｓｅ^ＴＭ振動多孔性プレート噴霧器（ＡｅｒｏＧｅｎ，Ｉｎｃ．，Ｓｕｎｎｙｖａｌｅ，Ｇａｌｉｆｏｒｎｉａ）、Ｓｉｄｅｓｔｒｅａｍ（登録商標）噴霧器（Ｍｅｄｉｃ−Ａｉｄ　Ｌｔｄ．，Ｗｅｓｔ　Ｓｕｓｓｅｘ，Ｅｎｇｌａｎｄ）、Ｐａｒｉ　ＬＣ　Ｐｌｕｓ（登録商標）およびＰａｒｉ　ＬＣ　Ｓｔａｒ（登録商標）ジェット噴霧器（Ｐａｒｉ　Ｒｅｓｐｉｒａｔｏｒｙ　Ｅｑｕｉｐｍｅｎｔ，Ｉｎｃ．，Ｒｉｃｈｍｏｎｄ，Ｖｉｒｇｉｎｉａ）、およびＡｅｒｏｓｏｎｉｃ^ＴＭ（ＤｅＶｉｌｂｉｓｓ　Ｍｅｄｉｚｉｎｉｓｃｈｅ　Ｐｒｏｄｕｋｔｅ（Ｄｅｕｔｓｃｈｌａｎｄ）ＧｍｂＨ、Ｈｅｉｄｅｎ，Ｇｅｒｍａｎｙ）およびＵｌｔｒａＡｉｒｅ（登録商標）（Ｏｍｒｏｎ　Ｈｅａｌｔｈｃａｒｅ，Ｉｎｃ．，Ｖｅｒｎｏｎ　Ｈｉｌｌｓ，Ｉｌｌｉｎｏｉｓ）超音波噴霧器が挙げられる。
【００５３】
（ＩＩＩ．エアロゾルの薬物動態）
エリスロマイシルアミンの溶液をＩＶおよび吸入経路でラットに投与し、そして血漿および肺中の薬物濃度を測定した。これらの研究からのデータを図８および９に示す。吸入送達経路について２つの用量レベル１．７ｍｇ／ｋｇおよび０．７ｍｇ／ｋｇを選択し、単回静脈用量（２５ｍｇ／ｋｇ）に対して比較した。
【００５４】
ＩＶ（２５ｍｇ／ｋｇ）、吸入（１．７ｍｇ／ｋｇ）、および（０．７ｍｇ／ｋｇ）についての肺におけるエリスロマイシルアミンの用量正規化ＡＵＣは、それぞれ、２４．２１μｇ・ｈ／グラム、１０６７．８４μｇ・ｈ／グラムおよび８４８．３４μｇ・ｈ／グラムであった。従って、吸入経路を介して肺へエリスロマイシルアミンを直接投与することにより、達成された肺の薬物レベルは、静脈経路による送達よりもミリグラムベースで約４０倍高かった。従って、吸入による抗生物質療法は、経口経路またはＩＶ経路による処置よりもさらに有効であるはずである。
【００５５】
（ＩＶ．エアロゾールの有効性）
エリスロマイシルアミンは、静脈内投与およびエアロゾール投与の両方で非常に有効であった。試験した最低の用量（１日あたり１０ｍｇ／ｋｇ）で、静脈内エリスロマイシルアミンは、実施例７に示すように、Ｓ．ｐｎｅｕｍｏｎｉａの肺負荷を検出限界（１０ＣＦＵ／肺のグラム数）以下まで減じた。エアロゾールはまた、非常に有効であって（図１０を参照のこと）、５ｍｇ／ｍｌのエアロゾール溶液（計算した用量は１日あたり０．１３ｍｇ／ｋｇ）でＳ．ｐｎｅｕｍｏｎｉａの検出可能な回復しか伴わなかった。さらに、エリスロマイシルアミンは、３日間、単回毎日用量のために必要な濃度より大きい濃度で単回エアロゾール用量として投与された場合、非常に有効であった。０．１３ｍｇ／ｋｇの単回用量は、３日連続して、０．１３ｍｇ／ｋｇ（５ケタの大きさの低下）と比較して有効性が低かった（ＣＦＵ／グラムとして２ケタより小さい大きさの低下）。しかし、０．６７ｍｇ／ｋｇの単回用量によって、肺組織からの生物体のほぼ完全なクリアランスが達成された。この効果は複数の用量有効性に類似しており、このことは、この濃度では、第二および第三の用量によってほとんど価値が得られないことを示す（図１１を参照のこと）。
【００５６】
エアロゾル化されたエリスロマイシルアミンの薬物動態的な評価が示唆し、そして有効性のデータが示すのは、複数の毎日ＩＶ用量、経口用量、またはエアロゾール用量に対して等価な肺濃度が、単回エアロゾール用量によって達成され得ること、およびこの単回用量が、３回の毎日用量と同様の有効性に必要であるより約３〜５倍大きいということである。
【００５７】
（有用性）
本発明の有用性の１つの局面は、マクロライド系抗生物質（例えば、エリスロマイシルアミン）の少ない容積、高濃度の処方物が、適切な噴霧器とともに用いられ得、慢性気管支炎、気管支拡張症、および肺炎（マクロライド感受性細菌または他の感染によって起きる）を有する人の気管支内空隙に有効用量のエリスロマイシルアミンを送達することである。この処方物は安全でありそして非常にコスト効果的である。さらに、この処方物は、商業的流通に適切な保存期間を提供するために、耐性について制御されたｐＨを伴う、窒素環境下で保持され得る。
【００５８】
（実施例１）
（エリスロマイシルアミン塩の調製のための一般的手順：酢酸エリスロマイシルアミンの合成）
１０．０ｇ（１３．６ｍｍｏｌ）のエリスロマイシルアミンを含有する１００ｍＬのＭｅＯＨ（氷槽中で冷却した）の溶液に、１．５６ｍＬ（２７．２ｍｍｏｌ、２．０当量）の氷酢酸を滴下して加えた。この溶液を３０分間にわたって環境温度まで暖め、次いで減圧下で溶媒を除いた。Ｅｔ_２Ｏ（５０ｍＬ）を添加して、スラリーを濃縮した。これを繰り返して、白色粉末として１１．５２ｇ（９６．９％）の１水和酢酸エリスロマイシルアミンを得た；ＩＲ（ＫＢｒ，ｃｍ^−１）１７１８，１５６０，１４０６，１１６８，１０８０，１０５５，１０１２；^１ＨＮＭＲ（４００ＭＨｚ，ＣＤ_３ＯＤ）δ０．８９（ｔ，３Ｈ，Ｊ＝７．２Ｈｚ），１．０６−１．３２（ｍ，２７Ｈ），１．３５−１．４７（ｍ，４Ｈ），１．５２−１．６６（ｍ，３Ｈ），１．８５−２．０２（ｍ，８Ｈ），２．０３−２．２６（ｍ，２Ｈ），２．４５−２．４９（ｍ，１Ｈ），２．６６−２．７７（ｍ，５Ｈ），２．９１−３．０９（ｍ，３Ｈ），３．２１−３．４０（ｍ，６Ｈ），３．５８（ｄ，１Ｈ，Ｊ＝７．０Ｈｚ），３．６７（ｓ，１Ｈ），３．７８−３．８３（ｍ，２Ｈ），４．１０−４．１３（ｍ，１Ｈ），４．５９（ｄ，１Ｈ，Ｊ＝７．０Ｈｚ），４．８８−５．０１（ｍ，１２Ｈ）；ＭＳ　ｍ／ｚ　７３５．６（Ｍ^＋−２ＡｃＯＨ−２Ｈ_２Ｏ）；ＫＦ　２．３３％Ｈ_２Ｏ。
【００５９】
Ｃ_４１Ｈ_８０Ｎ_２Ｏ_１７の分析計算値：Ｃ，５６．４０；Ｈ，９．２４；Ｎ，３．２１。実測値：Ｃ，５６．３８；Ｈ，９．２１；Ｎ，３．１６。
【００６０】
（実施例２）
（硫酸エリスロマイシルアミンの合成）
１０．０ｇ（１３．６ｍｍｏｌ）のエリスロマイシルアミンを含有する１００ｍＬのＭｅＯＨ（氷槽中で冷却した）の溶液に、０．７３ｍＬ（１３．６ｍｍｏｌ、１．０当量）の濃硫酸を滴下して加えた。この溶液を３０分間にわたって環境温度まで暖め、次いで減圧下で溶媒を除いた。Ｅｔ_２Ｏ（５０ｍＬ）を添加して、スラリーを濃縮した。これを繰り返して、白色粉末として１１．１３ｇ（９６．１％）の１水和硫酸エリスロマイシルアミンを得た；ＩＲ（ＫＢｒ，ｃｍ^−１）１７１８，１３８４，１１６８，１１２２，１０７８，１０１２；^１ＨＮＭＲ（４００ＭＨｚ，ＣＤ_３ＯＤ）δ０．８９（ｔ，３Ｈ，Ｊ＝７．２Ｈｚ），１．０８−１．３２（ｍ，２７Ｈ）、１．４５−１．６３（ｍ，７Ｈ），１．８９−２．０４（ｍ，２Ｈ）、２．２３−２．３１（ｍ，２Ｈ），２．４４−２．４７（ｍ，１Ｈ）、２．８４−２．８９（５Ｈ）、２．９９−３．０７（ｍ，３Ｈ），３．３０−３．４９（ｍ，６Ｈ），３．５８（ｄ，１Ｈ，Ｊ＝７．０Ｈｚ），３．６９（ｓ，１Ｈ），３．７８−３．８６（ｍ，２Ｈ），４．０９−４．１１（ｍ，１Ｈ），４．６０（ｄ，１Ｈ，Ｊ＝６．８Ｈｚ）、４．８７−４．９９（ｍ，１２Ｈ）；ＭＳ　ｍ／ｚ　７３５．７（Ｍ^＋−Ｈ_２ＳＯ_４−２Ｈ_２Ｏ）；ＫＦ　２．９３％Ｈ_２Ｏ。
【００６１】
Ｃ_３７Ｈ_７４Ｎ_２Ｏ_１６Ｓの分析計算値：Ｃ，５２．２２；Ｈ，８．７６；Ｎ，３．２９。実測値：Ｃ，５２．５５；Ｈ，８．９１；Ｎ，３．２７。
【００６２】
（実施例３）
（エリスロマイシルアミン塩酸塩の合成）
氷浴中で冷却したＭｅＯＨ１００ｍＬ中の１０．０ｇ（１３．６ｍｍｏｌ）のエリスロマイシルアミン（ｅｒｙｔｈｒｏｍｙｃｌａｍｉｎｅ）の溶液に、２．３４ｍＬ（２７．２ｍｍｏｌ，２．０ｅｑ）の３７％塩酸を滴下した。この溶液を、３０分間かけて周囲温度まで暖め、次いでこの溶媒を減圧下で除去した。Ｅｔ_２Ｏ（５０ｍＬ）を添加し、そしてスラリーを濃縮した。これを、白色粉末として１１．２４ｇ（９７．９％）のエリスロマイシルアミン塩酸塩二水和物を得るまで繰り返した；ＩＲ（ＫＢｒ，ｃｍ^−１）１７１８，１４６６，１３８３，１１７０，１０７８，１０５５，１０１１；^１Ｈ　ＮＭＲ（４００ＭＨｚ，ＣＤ_３ＯＤ）δ０．８７−０．９１（ｍ，３Ｈ），１．１０−１．３１（ｍ，２７Ｈ），１．４３−１．６５（ｍ，７Ｈ），１．８９−２．０１（ｍ，２Ｈ），２．２５−２．２７（ｍ，２Ｈ），２．４５−２．４８（ｍ，１Ｈ），２．８２−３．１０（ｍ，８Ｈ），３．３４−３．４２（ｍ，８Ｈ），３．５７−３．５８（ｍ，１Ｈ），３．６７（ｓ，１Ｈ），３．８０−３．８２（ｍ，２Ｈ），４．０８−４．１１（ｍ，１Ｈ），４．６１−５．００（ｍ，１３Ｈ）；ＭＳ　ｍ／ｚ　７３５．６（Ｍ^＋−２ＨＣｌ−２Ｈ_２Ｏ）；ＫＦ　４．３８％Ｈ_２Ｏ。
【００６３】
分析．Ｃ_３７Ｈ_７６Ｃｌ_２Ｎ_２Ｏ_１２についての計算値：Ｃ，５２．６５；Ｈ，９．０８；Ｎ，３．３２。実測値：Ｃ，５２．２１；Ｈ，９．１８；Ｎ，３．２０。
【００６４】
（実施例４）
（水性処方物およびエリスロマイシルアミン塩の安定性）
（溶液の調製）エリスロマイシルアミン（９．０ｇ，１２．２ｍＭ）遊離塩基を、風袋を計った１００ｍＬのＥｒｌｅｍｎｅｙｅｒフラスコに添加した。脱イオン水（２５ｍＬ）を、マグネチックスターラーで攪拌しながらこのフラスコに添加した。１Ｎの硫酸（２４．５ｍＬ，２当量）を、攪拌しながら徐々に添加した。この溶液が透明になったときに、このフラスコを攪拌プレートから取り除き、そして再秤量した。脱イオン水を滴下して、６２．９ｇの最終溶液重量を得た。この溶液を、３つの２０ｍＬの部分に分割し、そしてこのｐＨを、ｐＨメーターでモニタリングしながら、１Ｎの水酸化ナトリウムまたは硫酸の滴下によって所望の値（５．０、６．０、または７．０）に調整した。上記の手順を使用して、エリスロマイシルアミンの重量（６．０ｇおよび３．６ｇ）ならびに１Ｎの硫酸の容量（１６．３ｍＬおよび９．８ｍＬ）を調整することによって、１００ｍｇ／ｍＬおよび６０ｍｇ／ｍＬの溶液を調製した。
【００６５】
１５０、１００および６０ｍｇ／ｍＬのエリスロマイシルアミンの酢酸塩および塩酸塩の溶液を、１Ｎの酢酸および１Ｎの塩酸（２当量）を添加して塩を調製しそしてｐＨを調整することを除いて、上記のようにして調製した。
【００６６】
各濃度および各ｐＨでの各塩形態のアリコートを、４、４０、および６０℃、ならびに環境温度で貯蔵した。
【００６７】
（安定性の決定）全ての溶液を、調製の直後（ｔ＝０）、調製の２４時間後、４８時間後、８日後、１５日後、および２２日後に分析した（ただし、８日目に実質的に分解したサンプルは、引き続く分析から除外した）。
【００６８】
冷蔵したサンプルおよび加熱したサンプルを、サンプル調製の前に、少なくとも１時間にわたって環境温度まで平衡化した。全てのサンプルについての最終希釈容量は、１０ｍＬであった。全てのサンプルについての希釈剤は、５０ｍＭのリン酸緩衝液（ｐＨ６．５）とアセトニトリルとの８０：２０（ｖ／ｖ）の混合物からなる。
【００６９】
適切な量のサンプル（１５０ｍｇ／ｍＬの溶液については４０マイクロリットル、１００ｍｇ／ｍＬについては５０マイクロリットル、または６０ｍｇ／ｍＬの溶液については１００マイクロリットル）を、２０ｍＬのシンチレーションバイアルに移した。１０ｍＬの希釈剤をこのバイアルに添加し、そして徹底的に混合した。
【００７０】
（標準物質の調製）標準物質を二連で調製し、そして最大３日間で使用した。Ｅｒｙ−アミン遊離塩基（３０ｍｇ）を、風袋を計った５０ｍＬのメスフラスコに移し、そして正確な重量を記録した。サンプル希釈剤（４５ｍＬ）を添加し、そして溶解するまで簡単に超音波処理した。この標準物質を冷却し、そして希釈剤を用いて容量まで希釈した。
【００７１】
（サンプルおよび標準物質の分析）サンプルおよび標準物質を、逆相高速液体クロマトグラフィーによって分析した。５ミクロンの粒子サイズの２５０×４．６ｍｍのＰｈｅｎｏｍｅｎｅｘ　Ｌｕｎａ　ＣＮカラムを使用して、分離を行った。全ての分析を、Ａｇｉｌｅｎｔ　Ｔｅｃｈｎｏｌｏｇｉｅｓ　ＨＰ　１１００　クロマトグラフィーシステムで行い、そしてＡｇｉｌｅｎｔ　Ｔｅｃｈｎｏｌｏｇｉｅｓ　ＣｈｅｍＳｔａｔｉｏｎデータシステムを使用してデータを収集し、そして記録した。分析パラメーターは、以下の表２に示す通りであった。
【００７２】
（表２）
流速　　　　　１．０ｍＬ／分
カラム温度　　３０℃
注入容量　　　２０μＬ
検出器　　　　ＵＶ吸光度（２００ｎｍ）
実行時間　　　１０分
移動相Ａ　　　５０ｍＭホスフェート　ｐＨ２．１
移動相Ｂ　　　アセトニトリル
組成　　　　　８０／２０　Ａ／Ｂ。
【００７３】
（実施例５）
（エリスロマイシルアミン塩溶液の浸透圧重量モル濃度）
３部分のエリスロマイシルアミンＨＣｌ塩（０．６ｇ，１．０ｇおよび１．５ｇ）を、別個の１０ｍＬメスフラスコに計り入れた。Ｅａｓｙｐｕｒｅ　ＵＶ水（８ｍＬ）を各フラスコに添加し、そして完全に溶解するまで超音波処理し、次いで容量まで希釈した。この手順を、エリスロマイシルアミン硫酸塩および酢酸塩について繰り返した。各溶液のｐＨおよび浸透圧重量モル濃度を測定し、そして測定した浸透圧重量モル濃度を理論値と比較した。
【００７４】
実施例４で調製した塩（４℃）を、室温まで平衡化させ、そして浸透圧重量モル濃度を測定した。結果を表３に示す。
【００７５】
（表３）
【００７６】
【表２】

（実施例６）
（ラットへのエリスロマイシルアミンのエアロゾル送達：）
（エアロゾルの薬物動力学の特徴付け）
（静脈内の薬物動力学）：エリスロマイシルアミン（２５０ｍｇ）を、５ｍＬのＤＩ水に溶解し、そして１２ｍＬの濃硫酸を添加した。この溶液に希硫酸の溶液（１：１０　ｖ／ｖ）を徐々に添加して、薬物を完全に溶解した。希硫酸の溶液を徐々に添加して、溶液のｐＨを６．８〜７．２にした。ＤＩ水の添加によって、溶液の総容量を８ｍＬにした。エリスロマイシルアミン硫酸塩の溶液２００μｌ（２５ｍｇ／ｋｇ）を、側方尾静脈を介する静脈内投与によって、オスのＳｐｒａｇｕｅ−Ｄａｗｌｅｙラット（Ｓｉｍｏｎｓｅｎ　Ｌａｂｏｒａｔｏｒｉｅｓ，１１８０　Ｃ　Ｄａｙ　Ｒｏａｄ，Ｇｉｌｒｏｙ，ＣＡ　９５０２０）に送達した。動物を１〜４％のイソフルランで麻酔し、そして肺および血液のサンプルを、３匹のラットから、投薬の０．０８３、０．２５、０．５、１、２、４、６、８および２４時間後に収集した。血液サンプルを、抗凝固剤としてヘパリンを使用して、心臓の穿刺を介して収集した。肺を、血液のサンプリングの後に外科的に取り出し、そして気管支および気管を除去し、そして処分した。残った肺組織を、以下のように処理した。肺および血液のサンプルの両方を、すぐに氷上に置き、そして血液サンプルを収集の直後に遠心分離して、血漿サンプルを収集した。肺および血漿のサンプルの両方を、アッセイするまで−８０℃で保管した。
【００７７】
血漿および肺（肺組織１ｇ当たり）エリスロマイシルアミンの濃度を、確認されたＬＣ−ＭＳ法を使用して決定した。血漿サンプル（１００μｇ）を、抽出の前に、オレアンドマイシン（内部標準、１μｇ／ｍＬ）でスパイクした。血漿サンプル（１００μＬ）を、３．３％トリクロロ酢酸（ＴＣＡ）を用いてタンパク質を除去した。サンプルを遠心分離（１０，０００ｒｐｍ，１０分）し、そして遠心濾過（ｃｅｎｔｒｉｆｉｌｔｒａｔｉｏｎ）（１０，０００ｒｐｍ，１０分）のために、上清をＨＰＬＣ遠心濾過器（ｃｅｎｔｒｉｆｉｌｔｅｒ）に移した。移動相は、０．１％酢酸−アセトニトリル（７０：３０，ｖ／ｖ，ｐＨ＝３．２）溶液（０．５ｍｌ／分の流速で３分間）、続いて０．１％酢酸−アセトニトリル（６０：４０，ｖ／ｖ）（０．８ｍｌ／分の流速で３分間）からなる。ステンレス鋼の分析用カラム（Ｚｏｒｂａｘ　ＳＢ−Ｃ１８，２．１ｍｍ　ＩＤ×１５０．０ｍｍ，５μｍおよびＰｈｅｎｏｍｅｎｅｘカートリッジガードカラム）を、固定相として使用した。カラム温度は、５０℃であった。エリスロマイシルアミンの定量化を、ＨＰ　１１００　ＬＣ／ＭＳＤ　ＡＰＩ　Ｅｌｅｃｔｒｏｓｐｒａｙ　Ｓｙｓｔｅｍを使用して行った。データの取得を、選択的イオンモニタリングモードに設定した。この方法は、０．０１〜５０μｇ／ｍＬの濃度範囲において、線形（ｒ＞０．９９９０）であった。絶対回復（ａｂｓｏｌｕｔｅ　ｒｅｃｏｖｅｒｙ）は、９５．０±２．１９％であった。
【００７８】
肺サンプルを、ＤＩ水を用いてホモジナイズした。内部標準として、オレアンドマイシンをこのサンプルに添加した。このホモジネートを、０．９ＭのＴＣＡを用いてタンパク質を除去した。サンプルを、１０，０００ｒｐｍで１０分間遠心分離し、そして遠心濾過のために、この上清をＨＰＬＣ遠心濾過器に移した。移動相は、０．１％酢酸−アセトニトリル（７０：３０，ｖ／ｖ，ｐＨ＝３．２）（０．５ｍｌ／分の流速で３分間）、続いて０．１％酢酸−アセトニトリル（６０：４０，ｖ／ｖ）（０．８ｍｌ／分の流速で３分間）からなった。ステンレス鋼の分析用カラム（Ｚｏｒｂａｘ　ＳＢ−Ｃ１８，２．１ｍｍ　ＩＤ×１５０．０ｍｍ，５μｍおよびＰｈｅｎｏｍｅｎｅｘカートリッジガードカラム）を、固定相として使用した。カラム温度は５０℃であった。エリスロマイシルアミンの定量化を、ＨＰ　１１００　ＬＣ／ＭＳＤ　ＡＰＩ−Ｅｌｅｃｔｒｏｓｐｒａｙ　Ｓｙｓｔｅｍを使用して行った。データの取得を、選択的イオンモニタリングモードに設定した。このアッセイの直線性（ｒ＞０．９９９０）は、０．１〜２００μｇ／ｇの範囲であった。抽出効率は、９３．８±２．５４％であった。
【００７９】
薬物動態学的パラメーター、曲線下面積（ＡＵＣ）および平均滞留時間（ＭＲＴ）を、ＷｉｎＮｏｎｌｉｎ^ＴＭ　Ｐｒｏｆｅｓｓｉｏｎａｌ　Ｖｅｒｓｉｏｎ　２．０ソフトウエア（Ｐｈａｒｓｉｇｈｔ　Ｃｏｒｐｏｒａｔｉｏｎ）を使用して、統計学的モーメント理論に基づいて推定した。ピーク濃度（Ｃ_ｍａｘ）は推定しなかったが、観察した。
【００８０】
（吸入の薬物動力学）：６０ｍｇ／ｍＬの溶液について、５０ｍｌのメスフラスコ中の４３ｍＬのＤＩ水および４．２７ｍＬ（４．２７ｍｍｏｌ）の１Ｍ硫酸に、３．１９１ｇ（４．０８ｍｍｏｌ）のエリスロマイシルアミン（純度９４％）を添加した。次いで、この溶液を、さらなる５３μＬ（０．０５３ｍｍｏｌ）の１Ｍ硫酸の添加によってｐＨ６．５に調整した。さらなるＤＩ水を用いて、容量を５０ｍＬにした。６０ｍｇ／ｍＬ溶液を通常の生理食塩水で１／２に希釈することによって、３０ｍｇ／ｍＬの溶液を作製した。得られた溶液の浸透圧重量モル濃度は、Ｔｈｅ　Ａｄｖａｎｃｅｄ^ＴＭ　Ｍｉｃｒｏ−Ｏｓｍｏｍｅｔｅｒモデル３３００（Ａｄｖａｎｃｅｄ　Ｉｎｓｔｒｕｍｅｎｔｓ，Ｉｎｃ．，Ｎｏｒｗｏｏｄ，Ｍａｓｓ．）を使用して決定した場合、１４８ｍＯｓｍであった。
【００８１】
ラットを、３２ポートの鼻部のみのげっ歯類曝露システム（３２−ｐｏｒｔ　ｎｏｓｅ−ｏｎｌｙ　ｒｏｄｅｎｔ　ｅｘｐｏｓｕｒｅ　ｓｙｓｔｅｍ）（Ｂａｔｔｅｌｌｅ，Ｒｉｃｈｌａｎｄ，ＷＡ）において吸入を介して３０分間、エリスロマイシルアミン硫酸塩の３０ｍｇ／ｍＬまたは６０ｍｇ／ｍＬの溶液のいずれかに１度曝露した。このＢａｔｔｅｌｌｅシステムの鼻部のみのげっ歯類曝露システムは、Ｃａｎｎｏｎ　Ｆｌｏｗ−Ｐａｓｔ　Ｎｏｓｅ　ｏｎｌｙシステム（Ａｍ．Ｉｎｄ　Ｈｙｇ　Ａｓｓｏｃ　Ｊ　１９８３　Ｄｅｃ；４４（１２）９２３−８）に基づき、そして合計３２ポートの４つの積み重ね可能なステンレス鋼の層で構成される。このシステムは、入口流および排出流のモニタリングおよび制御を備え、エアロゾルデータは、Ｂａｔｔｅｌｌｅによって提供されるＮＯＲＥＳバージョン１．１．４ソフトウエアを使用して収集された。エリスロマイシルアミン溶液を、ＰＡＲＩ　ＬＣ　ＳＴＡＲ^ＴＭネブライザーを使用してエアロゾル化した。平均エアロゾル濃度を、曝露開始の１０および２０分後に採取したフィルターサンプルの重量分析によって決定した。平均エアロゾル濃度は、３０ｍｇ／ｍＬ溶液および６０ｍｇ／ｍＬ溶液についてそれぞれ、０．５４±０．０６および１．３６±０．３０ｍｇ／Ｌであった。
【００８２】
肺および血液のサンプルを、上記のように投薬の０．０８３、０．２５、０．５、１、２、４、８および２４時間後に３匹のラットから収集した。この吸入研究についてのサンプルの収集および取り扱い手順は、静脈内研究についてと同様であった。
【００８３】
吸入研究についての生物分析（ｂｉｏａｎａｌｙｔｉｃａｌ）アッセイの手順は、静脈内研究についての手順と同様であった。肺における算出した沈着した用量（肺用量）は、３０ｍｇ／ｍＬまたは６０ｍｇ／ｍＬのエリスロマイシルアミン溶液の３０分にわたる吸入用量後、それぞれ、約０．７０ｍｇ／ｋｇまたは１．７７ｍｇ／ｋｇであった。肺における肺用量を、以下のように算出した：
ＬＤＤ＝ＭＡＣ×ＭＶ×ＤＥ×ＦＬＤ÷ＭＢＷ
ここで、
ＬＤＤ＝肺に沈着した用量
ＭＡＣ＝平均エアロゾル濃度＝　３０および６０ｍｇ／ｍＬの溶液について、それぞれ、０．５４および１．３６ｍｇ／Ｌ
ＭＶ＝毎分換気量＝０．１Ｌ／分
ＤＥ＝曝露の持続時間＝３０分
ＦＬＤ＝肺沈着の画分＝０．１
ＭＢＷ＝平均体重＝０．２３ｋｇ。
【００８４】
エリスロマイシルアミンの静脈内投与および吸入投与後の肺における薬物動態学的パラメーターを、以下の表４に要約する：
（表４）
（ラットにおける静脈内投薬または２つの吸入投与後の、肺におけるエリスロマイシルアミンの薬物動態学的パラメーター（Ｎ＝３））
【００８５】
【表３】

１．投薬後０〜２４時間で見積られる曲線下面積
２．１ｍｇ／ｋｇに対して用量規格化された曲線下面積
３．投薬後０〜２４時間で見積られる平均滞留時間
ｎ．ｅ．：見積りされず。
【００８６】
（実施例７）
（感染のＳ．Ｐｎｅｕｍｏｎｉａラット肺モデルにおけるエリスロマイシルアミンのエアロゾルおよび静脈内（ＩＶ）での効力）
（方法）
雄性Ｓｐｒａｇｕｅ−Ｄａｗｌｅｙラットを、寒天ビーズ中に調製された５０〜１００マイクロリットルのＳ．Ｐｎｅｕｍｏｎｉａ　Ａ６６（株番号ＰＧＯ４７１６）を気管内投与することによって感染させた。この接種材料を、融解寒天中のＰＧＯ４７１６のブロス培養物に懸濁し、この寒天懸濁物を無菌鉱油中に混合しつつ懸濁して、この細菌を含有した寒天の小さなビーズを生成することによって調製した。このビーズを遠心分離によって回収し、無菌生理食塩水に再懸濁し、そして気管切開を介した肺への直接注射によって各動物に投与した。
【００８７】
エリスロマイシルアミン溶液を、無菌生理食塩水中に調製する。抗生物質を、尾部静脈への静脈内注射によってか、またはエアロゾル曝露によってかのいずれかで投与した。このエアロゾル曝露を、Ｉｎ−Ｔｏｘ　Ａｅｒｏｓｏｌ　Ｅｘｐｏｓｕｒｅ　Ｓｙｓｔｅｍ（モデル番号０４−１１００）を使用して、鼻部のみの曝露によって達成した。このシステムは、１つの端（鼻部ポート）においてこの系に対して開放しており、そしてシステム保全を維持するために残りの端において密閉されている、プラスチィックチューブに固定されたげっ歯類を曝露するために設計された、閉鎖エアロゾル送達システムである。このエアロゾルは、約６．５リットル／分の流量でＰａｒｉ　ＬＣ　Ｓｔａｒ^ＴＭエアジェットネブライザーによって生成される。吸引を９リットル／分に設定し、その結果、より希薄な空気を用いてこのシステムを通過する総流量は、７．５リットル／分である。
【００８８】
処置は、感染後２４時間にて開始され、そして１日につき１回で、３日間継続する。エアロゾルを、毎日３０分間投与した。感染後第４日目かつ最終の投薬後１２時間において、動物を屠殺し、そして肺を外科的に取り出した。この切除の後、肺をホモジェナイズし、希釈し、定量的に血液寒天上にプレートした。プレートを、２４時間インキュベートし、そしてＳ．ｐｎｅｕｍｏｎｉａｅのコロニーを計数して、細菌の取り込みを決定する。この結果を表５に示す：
（表５）
（ラットＰｎｅｕｍｏｎｉａｅモデルにおけるＳ．ｐｎｅｕｍｏｎｉａｅに対するエリスロマイシルアミンの効力）
【００８９】
【表４】

＊ＢＱＬ＝定量の限界より下
（実施例８）
（エリスロマイシルアミンの単回投薬処置後の感染のＳ．Ｐｎｅｕｍｏｎｉａラット肺モデルにおけるエリスロマイシルアミンのエアロゾルの効力）
雄性Ｓｐｒａｇｕｅ−Ｄａｗｌｅｙラットを、実施例７に記載のように、感染させ、そしてエアロゾル処置に曝露した。この単回処置を、３０分間にわたって示される用量で投与されたエアロゾルにて感染後２４時間で開始した。さらなる処置を行わず、そして動物を外科手術まで観察した。感染後第４日目（投薬後、第３日目）において、この動物を屠殺し、これらの肺を外科的に取り出した。この切除の後、肺をホモジェナイズし、希釈し、定量的に血液寒天上にプレートした。プレートを、２４時間インキュベートし、そしてＳ．ｐｎｅｕｍｏｎｉａｅのコロニーを計数し、細菌の取り込みを決定する。単回用量投与後の結果を、図１１に示す。さらなる結果を、表６に示す：
（表６）
（ラットＰｎｅｕｍｏｎｉａｅモデルにおけるＳ．ｐｎｅｕｍｏｎｉａｅに対するエリスロマイシルアミンの効力）
【００９０】
【表５】

ＢＱＬ＝定量の限界より下。
【００９１】
（実施例９）
（イヌへのエリスロマイシルアミンのエアロゾル送達：）
（エアロゾルの薬物動力学の特徴付け）
（吸入の薬物動力学：）
６０ｍｇ／ｍＬの溶液について、５０ｍｌのメスフラスコ中の４３ｍＬのＤＩ水および４．２７ｍＬ（４．２７ｍｍｏｌ）の１Ｍ硫酸に、３．１９１ｇ（４．０８ｍｍｏｌ）のエリスロマイシルアミン（純度９４％）を添加した。次いで、この溶液を、さらなる５３μＬ（０．０５３ｍｍｏｌ）の１Ｍ硫酸の添加によってｐＨ６．５に調整した。さらなるＤＩ水を用いて、容量を５０ｍＬにした。イヌを、吸入マスク曝露システム（Ｉｎｖｅｒｅｓｋ　Ｒｅｓｅａｒｃｈ，Ｓｃｏｔｌａｎｄ，ＵＫ）を介して３０分間にわたって、エリスロマイシルアミン硫酸塩の６０ｍｇ／ｍＬ溶液に１度曝露した。
【００９２】
これらのイヌを、イヌ保有領域にあるその囲いから連れ出し、そして投薬実験室に移した。投薬の間、これらの動物を、動物係員によってか、または、吊なわ／ハーネスシステムのいずれかで拘束した。吸入投薬は、投薬の開始に先だって適切に特徴付けられたネブライザーに接続し密閉フェイスマスクを使用して行われた。この投薬装置は、可撓性のあるチューブに付着されたフェイスマスクおよびマウスピースを組み込み、このチューブはネブライザーデバイスに接続された。このマウスピースを、これらの動物の口腔内の舌上に位置付け、そしてゴム製のスリーブによってこのフェイスマスクをイヌ鼻先の周りで密閉した。このマスクからの排気バルブを、排気システムに接続した。この投薬装置が完全に組立てられ、そしてイヌに適合された場合、吸気が、可撓性チューブを通過するイヌへのエアロゾルの動きによって示される。
【００９３】
肺サンプルを、投薬後、２時間、２４時間、４８時間、７２時間、９６時間、および１２０時間において、２匹のイヌから収集した。肺を、このイヌから外科的に切除し、各葉（右尾方（右下葉）、左尾方（左下葉）、右頭方（右上葉）、左頭方（左上葉）、右中央（右中葉）、および副葉）を、アッセイのために分離した。血漿サンプルを、２時間、２４時間後、４８時間後、７２時間後、９６時間後、および１２０時間後に、全ての生存している動物から収集した。
【００９４】
血漿および肺（肺組織１ｇ当たり）のエリスロマイシルアミンの濃度を、ＬＣ−ＭＳ法を使用して決定した。血漿サンプル（１００μｇ）を、抽出の前に、オレアンドマイシン（内部標準、１μｇ／ｍＬ）でスパイクした。血漿サンプル（１００μＬ）を、３．３％トリクロロ酢酸（ＴＣＡ）を用いてタンパク質を除去した。サンプルを遠心分離（１０，０００ｒｐｍ，１０分）し、そして遠心濾過（ｃｅｎｔｒｉｆｉｌｔｒａｔｉｏｎ）（１０，０００ｒｐｍ，１０分）のために、上清をＨＰＬＣ遠心濾過器（ｃｅｎｔｒｉｆｉｌｔｅｒ）に移した。移動相は、０．１％酢酸−アセトニトリル（７０：３０，ｖ／ｖ，ｐＨ＝３．２）溶液（０．５ｍｌ／分の流速で３分間）、その後０．１％酢酸−アセトニトリル（６０：４０，ｖ／ｖ）（０．８ｍｌ／分の流速で３分間）からなる。ステンレス鋼の分析用カラム（Ｚｏｒｂａｘ　ＳＢ−Ｃ１８，２．１ｍｍ　ＩＤ×１５０．０ｍｍ，５μｍおよびＰｈｅｎｏｍｅｎｅｘカートリッジガードカラム）を、固定相として使用した。カラム温度は、５０℃であった。エリスロマイシルアミンの定量化を、ＨＰ　１１００　ＬＣ／ＭＳＤ　ＡＰＩ　Ｅｌｅｃｔｒｏｓｐｒａｙ　Ｓｙｓｔｅｍを使用して行った。データの取得を、選択的イオンモニタリングモードに設定した。この方法は、０．０１〜５０μｇ／ｍｌの濃度範囲において、線形（ｒ＞０．９９９０）であった。絶対回復（ａｂｓｏｌｕｔｅ　ｒｅｃｏｖｅｒｙ）は、９０％を超えた。
【００９５】
肺サンプルを、ＤＩ水を用いてホモジナイズした。内部標準として、オレアンドマイシンをこのサンプルに添加した。このホモジネートから、０．９ＭのＴＣＡを用いてタンパク質を除去した。サンプルを、１０，０００ｒｐｍで１０分間遠心分離し、そして遠心濾過のために、この上清をＨＰＬＣ遠心濾過器に移した。移動相は、０．１％酢酸−アセトニトリル（７０：３０，ｖ／ｖ，ｐＨ＝３．２）（０．５ｍｌ／分の流速で３分間）、その後０．１％酢酸−アセトニトリル（６０：４０，ｖ／ｖ）（０．８ｍｌ／分の流速で３分間）からなった。ステンレス鋼の分析用カラム（Ｚｏｒｂａｘ　ＳＢ−Ｃ１８，２．１ｍｍ　ＩＤ×１５０．０ｍｍ，５μｍおよびＰｈｅｎｏｍｅｎｅｘカートリッジガードカラム）を、固定相として使用した。カラム温度は５０℃であった。エリスロマイシルアミンの定量化を、ＨＰ　１１００　ＬＣ／ＭＳＤ　ＡＰＩ−Ｅｌｅｃｔｒｏｓｐｒａｙ　Ｓｙｓｔｅｍを使用して行った。データの取得を、選択的イオンモニタリングモードに設定した。このアッセイの線形性（ｒ＞０．９９）は、肺については、２〜１００μｇ／ｇの範囲であった。抽出効率は、９０％を超えた。
【００９６】
薬物動態学的パラメーター、曲線下面積（ＡＵＣ）および平均滞留時間（ＭＲＴ）ならびに半減期（Ｔ_１／２）を、ＷｉｎＮｏｎｌｉｎ^ＴＭ　Ｐｒｏｆｅｓｓｉｏｎａｌ　Ｖｅｒｓｉｏｎ　３．１ソフトウエア（Ｐｈａｒｓｉｇｈｔ　Ｃｏｒｐｏｒａｔｉｏｎ）を使用して、統計学的モーメント理論に基づいて推定した。ピーク濃度（Ｃ_ｍａｘ）は見積らなかったが、観察した。
【００９７】
イヌにおける、エリスロマイシルアミンの投与後の肺吸入および血漿吸入における薬物動態学的パラメーターは、以下の表７、ならびに図１３および図１４で要約される：
（表７）
（イヌにおける６０ｍｇ／ｍＬ溶液の３０分間吸入投与後の、肺および血漿におけるエリスロマイシルアミンの薬物動態学的パラメーター（Ｎ＝２））
【００９８】
【表６】

１．平均滞留時間
ｎ．ｅ．：見積りされず。
【００９９】
（実施例１０）
（エリスロマイシルアミンの液体エアロゾル送達）
１／４（ｑｕａｔｅｒ）標準生理食塩水（ｐＨ７．０）中のエリスロマイシルアミン硫酸塩の溶液（１００ｍｇ／ｍＬ）を、先述した実施例の一般的な手順に従って調製する。この溶液の１．０ｍＬ用量を、急性増悪期にある慢性気管支炎（ＡＥＣＢ）に罹患するヒト被験体へ、ＡｅｒｏＧｅｎ　Ａｅｒｏｄｏｓｅ^ＴＭ吸入器を用いて１０分未満のエアロゾル吸入によって投与する。ＡＥＣＢおよびＡＥＣＢの症状に関与する細菌の低減が観察される。
【０１００】
（実施例１０）
（エリスロマイシルアミンの乾燥粉末エアロゾル送達）
エリスロマイシルアミン硫酸塩の乾燥粉末処方物（１００ｍｇ）および乾燥粉末キャリア（等部の、ラクトース、２−ヒドロキシプロピル−β−シクロデキストリン、マンニトール、およびアスパルテーム；総質量２５ｍｇ）を、調製する。この処方物を、急性増悪期にある慢性気管支炎（ＡＥＣＢ）に罹患するヒト被験体に、Ｇｌａｘｏ　Ｖｅｎｔｏｌｉｎ　Ｒｏｔｏｈａｌｅ^ＴＭ吸入器を用いて２分間未満のエアロゾル吸入によって投与する。ＡＥＣＢおよびＡＥＣＢの症状に関与する細菌の低減が観察される。
【０１０１】
本発明の好ましい実施形態が例示され、そして記載されるが、種々の変化が、本発明の精神および範囲から逸脱することなしに、それらにおいてなされ得ることが理解される。
【図面の簡単な説明】
本発明の上記の局面および付属する利点の多くは、本発明が、添付の図面とともに解釈されたときに、上記の詳細な説明を参照することによってより良く理解される場合に、より容易に理解される。
【図１】
図１は、エリスロマイシルアミンおよびジリスロマイシンの化学構造を示す。
【図２】
図２は、実施例４に記載されるような、４℃における６０ｍｇ／ｍＬ、１００ｍｇ／ｍＬおよび１５０ｍｇ／ｍＬ、かつｐＨ５．０、ｐＨ６．０およびｐＨ７．０の水溶液中での塩酸エリスロマイシルアミンの安定性のグラフ表示である。
【図３】
図３は、実施例４に記載されるような、２５℃における６０ｍｇ／ｍＬ、１００ｍｇ／ｍＬおよび１５０ｍｇ／ｍＬ、かつｐＨ５．０、ｐＨ６．０およびｐＨ７．０の水溶液中での塩酸エリスロマイシルアミンの安定性のグラフ表示である。
【図４】
図４は、実施例４に記載されるような、４０℃における６０ｍｇ／ｍＬ、１００ｍｇ／ｍＬおよび１５０ｍｇ／ｍＬ、かつｐＨ５．０、ｐＨ６．０およびｐＨ７．０の水溶液中での塩酸エリスロマイシルアミンの安定性のグラフ表示である。
【図５】
図５は、実施例４に記載されるような、６０℃における６０ｍｇ／ｍＬ、１００ｍｇ／ｍＬおよび１５０ｍｇ／ｍＬ、かつｐＨ５．０、ｐＨ６．０およびｐＨ７．０の水溶液中での塩酸エリスロマイシルアミンの安定性のグラフ表示である。
【図６】
図６は、実施例４に記載されるような、６０℃における６０ｍｇ／ｍＬ、１００ｍｇ／ｍＬおよび１５０ｍｇ／ｍＬ、かつｐＨ５．０、ｐＨ６．０およびｐＨ７．０の水溶液中での硫酸エリスロマイシルアミンの安定性のグラフ表示である。
【図７】
図７は、実施例４に記載されるような、６０℃における６０ｍｇ／ｍＬ、１００ｍｇ／ｍＬおよび１５０ｍｇ／ｍＬ、かつｐＨ５．０、ｐＨ６．０およびｐＨ７．０の水溶液中での酢酸エリスロマイシルアミンの安定性のグラフ表示である。
【図８】
図８は、実施例６に記載されるような、２５ｍｇ／ｋｇの単回静脈内用量または３０ｍｇ／ｍｌ溶液または６０ｍｇ／ｍｌ溶液の３０分間にわたる単回吸入用量（０．７ｍｇ／ｋｇ肺用量または１．７７ｍｇ／ｋｇ肺用量）後の、ラット（ｎ＝３）中でのエリスロマイシルアミンの平均血漿濃度を示す。
【図９】
図９は、実施例６に記載されるような、２５ｍｇ／ｋｇの単回静脈内用量または３０ｍｇ／ｍｌ溶液または６０ｍｇ／ｍｌ溶液の３０分間にわたる単回吸入用量（０．７ｍｇ／ｋｇ肺用量または１．７７ｍｇ／ｋｇ肺用量）後の、ラット（ｎ＝３）中でのエリスロマイシルアミンの平均肺濃度を示す。
【図１０】
図１０は、実施例７に記載されるような、ラット（ｎ＝３）に対して、５ｍｇ／ｍＬ（０．１３ｍｇ／ｋｇ）、２５ｍｇ／ｍＬ（０．２７ｍｇ／ｋｇ）および５０ｍｇ／ｍＬ（１．３ｍｇ／ｋｇ）吸入用量を含む１日３０分間の吸入投与を３日間行った後の、Ｓ．ｐｎｅｕｍｏｎｉａｅ肺感染モデルにおけるエリスロマイシルアミンの効力を示す。
【図１１】
図１１は、実施例８に記載されるような、ラット（ｎ＝３）に対して、１ｍｇ／ｍＬ（０．０３ｍｇ／ｋｇ）、５ｍｇ／ｍＬ（０．１３ｍｇ／ｋｇ）、２５ｍｇ／ｍＬ（０．２７ｍｇ／ｋｇ）および５０ｍｇ／ｍＬ（１．３ｍｇ／ｋｇ）吸入用量を含む単回用量として３０分間の吸入投与後の、Ｓ．ｐｎｅｕｍｏｎｉａｅ肺感染モデルにおけるエリスロマイシルアミンの効力を示す。
【図１２】
図１２は、実施例９に記載されるような、イヌ中で６０ｍｇ／ｍＬ硫酸塩溶液を、単回用量の３０分間吸入投与した後の、エリスロマイシルアミンの平均血漿濃度および平均全肺濃度を示す。
【図１３】
図１３は、実施例９に記載されるような、イヌ中で６０ｍｇ／ｍＬ硫酸塩溶液を、単回用量の３０分間吸入投与した後の、個々の肺葉におけるエリスロマイシルアミンの平均肺濃度を示す。[0001]
(Field of the Invention)
The present invention relates to novel and improved macrolide formulations for delivery by inhalation (eg, erythromycylamine formulations), and improvements for the treatment of susceptible acute or chronic endobronchial infections Related to the way it was done. In particular, the invention relates to formulations comprising at least one concentrated macrolide antibiotic in a physiologically acceptable liquid solution or dry powder form. These formulations, in liquid aerosol or dry powder form, are suitable for the delivery of macrolide antibiotics (eg, erythromycilamine) to the intrabronchial airway space of the lung, where aerosolized droplets of the formulation or A substantial portion of the particles have a median aerodynamic diameter of between 1-5 μm. Caused effective amount of macrolide formulated and aerosol delivery include acute and chronic endobronchial infections and pneumonia (especially, Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Moraxella catarrhalis, Legionella pneumonia, by Chlamydia pneumoniae, and Mycoplasma pneumoniae Are effective in the treatment and / or prevention of The new formulation delivers a small but effective amount of a macrolide antibiotic to the site of infection. In yet another aspect, the present invention relates to new and improved unit dose formulations of macrolide antibiotics for delivery by aerosol inhalation.
[0002]
(Background of the Invention)
Streptococcus pneumonia and other typical and atypical pathogens infect the endobronchial space in the lungs of individuals with chronic obstructive pulmonary disease (COPD) [S. Chodosh et al., Clinical Infectious Diseases 1998; 27: 730-738]. COPD is most commonly manifested as chronic bronchitis (CB) and emphysema.
[0003]
Chronic bronchitis is a lung disease characterized by inflammation and progressive destruction of lung tissue. Pulmonary weakness in CB patients is associated with chronic cough, increased daily sputum production, and accumulation of purulent sputum produced as a result of chronic intrabronchial infection caused by impaired lung function. Acute relapse of chronic bronchitis (AECB) often results in an increasing cough, purulent sputum production, and Streptococcus pneumonia, H. et al. Influenzae and Moraxella catarrhalis are characterized by clinical deterioration. Pneumonia can also result from infection by these organisms, either newly or as a complication of COPD. Despite the controversy over the adequacy of antimicrobial therapy for the treatment of CB, particularly acute relapse of CB, Saint et al. (JAMA 1995; 273: 957-960) have reported that oral antimicrobial therapy has Has proven to offer some clinical benefits. In addition, antimicrobial dose was important with respect to time to relapse. Thus, higher doses of oral antimicrobial agents were associated with a higher median infection-free interval (S. Chodoshi et al., Clinical Infectious Diseases 1998; 27: 730-738).
[0004]
Currently, oral administration of macrolide antibiotics and fluoroquinolones active against typical and atypical pathogens are selected for treatment of CB. However, oral administration of macrolide antibiotics has deleterious side effects. The most common side effects associated with oral / parenteral macrolide antibiotic treatment are diarrhea / loose bowel movements, nausea, abdominal pain and vomiting (RN Brogden D. Peters, Drugs, 1994; 48: 599). -616 and HD Langtry, RN Brogden Drugs 1997; 53: 973-1004 and references cited herein). In addition, pseudomembranous colitis is a serious side effect associated with oral antibiotic therapy, including oral macrolide therapy (SH Ahmad et al., Indian J. Pediatr. 1993, 60: 591-594). The penetration of macrolide into lung tissue after oral administration varies according to dose and composition (RN Brogden D. Peters, Drugs, 1994; 48: 599-616 and HD Langtry, RN. Brogden Drugs 1997; 53: 973-1004 and references cited herein). In addition, macrolides are associated with changes in systemic concentrations of unrelated drugs (eg, theophylline) due to interactions with the liver's cytochrome-based metabolic system. Such drug-drug interactions often require dosage adjustment or removal of one component from the treatment regimen.
[0005]
Erythromycylamine is a 14-membered ring macrolide belonging to the erythromycin family of antibiotics, and has a similar in vitro antibiotic spectrum to erythromycin A, and, like erythromycin A, has been shown to treat atypical and atypical pneumonia. It is an effective treatment. Erythromysylamine has a C-9 amino function with an S structure instead of the C-9 carbonyl group found in erythromycin A. One important limitation of erythromycylamine is its lack of oral absorption, and thus a prodrug (dirithromycin) was developed to achieve useful therapeutic concentrations. The prodrug of erythromysylamine is dirithromycin, which is characterized by a crosslinked acetal function between the C-9 amino group and the C-11 hydroxyl group (see FIG. 1). Cyclic acetals are rapidly hydrolyzed in plasma by a non-enzymatic process (half-life of about 30 minutes). Dilithromycin has been shown to successfully treat relapses that occur in patients with CB (M. Cazzola et al., Respiratory Medicine; 1998; 92: 895-901). A major advantage of erythromysylamine is its long half-life (30-44 hours) (RN Brogden D. Peters, Drugs, 1994; 48: 599-616). Unfortunately, oral bioavailability of dirithromycin has high excretion (62-81%) in feces, mainly as erythromycilamine, and is only 10-14% in humans. Because erythromycylamine is not absorbed and its prodrug (dirithromycin) is poorly absorbed, a limited amount of active drug is used systemically to treat lung infections caused by typical and atypical bacteria. Available. Sufficient erythromycylamine concentrates at the site of infection to provide a therapeutic effect, but the concentration of the drug is limited. Higher oral doses or more frequent dosing of dirithromycin increase drug concentrations at the site of action; however, increased adverse events appear to occur and may increase patient distress and complications.
[0006]
One of the first studies to use aerosolized antibiotics for the treatment of lung infection was reported in Lancet, 22: 1377-9 (1981). A controlled double-blind study on 20 CF patients demonstrated that aerosol administration of carbenicillin and aminoglycoside gentamicin could improve the health of CF patients. Since that time, sporadic reports in the literature have generally investigated aerosol delivery of aminoglycosides and especially tobramycin (see, eg, US Pat. No. 5,580,269). However, evaluation and comparison of these studies is often difficult due to differences in antibiotic formulations, respiratory techniques, nebulizers and compressors. In addition, aerosol delivery is often difficult to assess due to differences in formulation, aerosol delivery device, dosage, particle size, regimen, and the like. For example, if the median aerodynamic diameter (MMAD) of the size is greater than 5 μm, the particles will routinely deposit in the upper respiratory tract and reduce the amount of antibiotic delivered to the site of infection in the lower respiratory tract. Arch. Dis. Child. , 68: 788 (1993), emphasizes the need for standardized procedures and improvements in aerosol administration of drugs to CF patients.
[0007]
Effective aerosol administration is currently undermined by the absence of additives and the lack of physiologically compatible formulations, and especially by the inability of certain nebulizers to produce small and uniform particle sizes. I have. The range of aerosolized particle sizes required to deliver the drug to the endobronchial space, peri-lung, and site of infection is between about 1 μm and 5 μm. Many nebulizers that aerosolize therapeutic agents (including aminoglycosides) produce a large number of aerosol particles having a size of less than 1 μm or greater than 5 μm. To be therapeutically effective, many aerosolized antibiotic particles should not have a MMAD greater than 5 μm. If the aerosol contains a large number of particles with a MMAD greater than 5 μm, the larger size particles will deposit in the upper respiratory tract and reduce the amount of antibiotic delivered to the site of infection in the lower respiratory tract.
[0008]
Currently, three types of available nebulizers (jet nebulizer, vibrating porous plate nebulizer, and ultrasonic nebulizer) have aerosols with a diameter size between 1-5 μm (the preferred particle size for the treatment of bacterial infections of the lung). Particles can be created and delivered. Accordingly, it would be highly advantageous to provide a macrolide formulation that can be efficiently aerosolized in jet nebulizers, vibrating porous plate nebulizers, and ultrasonic nebulizers. Additionally, newer aerosol generation technologies are currently available, including mechanical extrusion and passive and active dry powder inhalers that are useful for delivery of therapeutic agents in dry powder form.
[0009]
Another requirement for an acceptable formulation is an appropriate shelf life. Generally, antibiotics, particularly antibiotic solutions for intravenous administration, contain phenol or other preservatives to maintain efficacy and minimize the formation of degradation products. However, phenols and other preservatives, when aerosolized, can induce bronchospasm, an unwanted occurrence in patients with pulmonary disease such as chronic bronchitis.
[0010]
Administration of macrolide antibiotics (e.g., erythromycylamine) for inhalation in the form of a liquid or dry powder aerosol has the advantage of overcoming the poor oral bioavailability associated with prodrugs; Provide an effective concentration of antibiotic to the lung that cannot be achieved by any of the intravenous routes. Further advantages of aerosol delivery of erythromycylamine are the inherently high affinity for lung tissue and resistance in the plasma compartment (long plasma / tissue half-life). The combination of high concentration aerosol delivery, long plasma / tissue half-life and high lung affinity allows for safer macrolide treatment, which eradicates or substantially eliminates endobronchial infection after administration of a single aerosol dose. Can be significantly reduced.
[0011]
Thus, at a pH adjusted to a level that slows or prevents degradation, it is preservative-free and is tolerated by patients and provides a suitable half-life suitable for commercial distribution, storage and use It would be highly advantageous to provide a macrolide antibiotic formulation (eg, an erythromycylamine formulation).
[0012]
Thus, for example, by using a jet nebulizer, a vibrating porous plate nebulizer, or an ultrasonic nebulizer, or a dry powder injector, the aerosol particles can be efficiently aerosolized, mainly to aerosol particle sizes in the range of 1-5 μm1. Providing concentrated formulations of macrolide antibiotics (e.g., erythromycylamine, erythromycin A, roxithromycin, azithromycin, and clarithromycin), comprising an effective concentration of the macrolide antibiotic in one form. Is an object of the present invention.
[0013]
(Summary of the Invention)
In accordance with the present invention, endobronchial infections (eg, the bacteria Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Moraxella catarrhonalis or atypical pathogens, or / and atypical pathogens caused by the atypical pathogens). Human and non-human animal subjects at risk can be treated with an antimicrobial effective amount of a macrolide antibiotic (eg, erythromycylamine, erythromycin A, roxithromycin) in a liquid solution or dry powder form suitable for aerosol generation. , Azithromycin, or clarithromycin It has now been discovered that by administration to a subject by inhalation, it can be treated effectively and efficiently.
[0014]
Accordingly, one aspect of the present invention is directed to macrolide antimicrobial drugs (eg, erythromycylamine, erythromycin A, roxithrothyl) into the endobronchial space of a subject suffering from or at risk of bacterial pulmonary infection. Mycin, azithromycin, and clarithromycin) in a concentrated formulation suitable for effective delivery by inhalation.
[0015]
Another aspect of the present invention is that the bacteria Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Moraxella catarrhalis, and / or the atypical pathogen Legionella pneumoniae, which are infected with Haemophilus influenzae, Staphylococcus aureus, and the atypical pathogens. The interior space provides a formulation suitable for efficient delivery of macrolide antimicrobial drugs (eg, erythromycylamine, erythromycin A, roxithromycin, azithromycin, and clarithromycin).
[0016]
Another aspect of the present invention is to treat the Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Moraxella catarrhalis, and / or the atypical pathogen Legionella pneumoniaa / neumoniaa / neumoniae / neumoniaia / neumoniae, with the atypical pathogens To prevent or substantially reduce the efficacy of macrolide antimicrobial drugs (eg, erythromycilamine, erythromycin A, roxithromycin, azithromycin, and clarithromycin) in a subject's endobronchial space Provide a formulation suitable for delivery.
[0017]
Yet another aspect of the invention is to equilibrate 0.5-5 ml of a physiologically acceptable carrier (e.g., saline diluted to one-fourth strength of normal saline). Provide a liquid formulation comprising 50-750 mg of a macrolide antimicrobial drug (e.g., erythromycylamine, erythromycin A, roxithromycin, azithromycin, and clarithromycin), wherein the formulation comprises It has an osmotic pressure, salinity, and pH that is resistant to and is suitable for delivery to a subject in concentrated form by aerosol inhalation.
[0018]
Yet another aspect of the present invention relates to an equivalent amount of 25-250 mg of a macrolide antimicrobial drug in a physiologically acceptable dry powder carrier for delivery to a subject in concentrated form by aerosol inhalation (E.g., erythromycylamine, erythromycin A, roxithromycin, azithromycin, and clarithromycin), wherein the dry powder formulation comprises about 50-90% by weight of macrolide Contains antibacterial drugs.
[0019]
Yet another aspect of the present invention relates to an antimicrobial effective amount of a macrolide antimicrobial drug (eg, erythromycylamine, erythromycin A, roxithromycin) formulated in a physiologically compatible liquid solution or dry powder form. Azithromycin, and clarithromycin) by administration to a subject in need of such treatment by inhalation of an aerosol formulation containing the same, thereby providing a method for the treatment of a lung infection caused by susceptible bacteria. Here, the median aerodynamic diameter (MMAD) of the size of the particles in the aerosol formulation is mainly between 1 and 5 μm.
[0020]
In another aspect, the present invention provides a unit dose formulation and unit dose device adapted for use in connection with a high efficiency inhalation system, wherein the unit dose device comprises a relatively small amount of a macrolide antibiotic of the present invention. It comprises a container designed to hold and store the substance formulation and deliver the formulation to an inhalation device for delivery to the subject in aerosol form. In one aspect, the unit dose device of the invention comprises less than about 2.0 ml of a liquid macrolide antibiotic formulation comprising about 50 to about 150 mg / ml of the macrolide antibiotic in a physiologically acceptable liquid carrier. , Including a sealed container (eg, an ampoule). Alternatively, the container of the unit dose device may contain less than about 1.5 ml or less than about 1.0 ml of the liquid macrolide antibiotic formulation, and the macrolide antibiotic formulation may contain from about 80 to about 180 mg / ml, Or it may contain about 90 to about 120 mg / ml of macrolide antibiotic. In another aspect, a unit dose device of the invention comprises a sealed, comprising a dry powder macrolide antibiotic formulation comprising from about 20 to about 250 mg of a macrolide antibiotic in a physiologically acceptable dry powder carrier. Container (eg, ampoule). The sealed unit dose container of the present invention is preferably adapted to deliver a macrolide antibiotic formulation to a high efficiency inhalation device for aerosolization and inhalation by a subject.
[0021]
(Detailed description of preferred embodiments)
Erythromycylamine and dirithromycin are macrolides having the chemical structure shown in FIG. Dilithromycin is a prodrug of erythromycilamine, a broad spectrum macrolide antibiotic used in the treatment of AECB and pneumonia. Macrolide antibiotics useful in the present invention include, for example, erythromycylamine, dirithromycin (prodrug of erythromycylamine), erythromycin A, clarithromycin (6-O-methylerythromycin), azithromycin, and roxithromycin Mycin is mentioned. Other newer macrolides (e.g., ketolides (e.g., ABT-773 (39 ^th ICAAC (1999), September 26-29, Abstract F-2133-2141) and HMR-3647 (Drugs of the Future, 23, 591 (1998), 38). ^th ICAAC (1998), September 24-27, Abstracts A-49), and anhydrolides (see J. Med. Chem. 1998, 41, 1651-1659 and 1660-1670) are also included in the present invention. Can be used in practice. In one aspect of the invention, the macrolide antibiotic used in the aerosol formulations described herein is erythromycylamine or dirithromycin. Erythromycylamine and dirithromycin have the chemical structures shown in FIG.
[0022]
In accordance with the present invention, there is provided a method for the treatment of a subject in need thereof (eg, a subject suffering from an endobronchial infection), the method comprising: Administering a large amount of a macrolide antibiotic formulation. This aspect of the invention provides a small volume respiratory fluid to produce the desired macrolide aerosol particle size of 1-5 μm to effectively deliver macrolide to the endobronchial space to treat susceptible microbial infections. It is particularly suitable for the formulation of concentrated macrolides (eg, erythromysylamine) for aerosolization by operative high power rates and high efficiency inhalers. The formulation preferably comprises a macrolide aerosol that is small in volume of a physiologically acceptable solution (eg, well tolerated by the patient but prevents undesirable side effects (eg, bronchospasm and cough)). Containing a minimum but still effective amount of macrolide formulated in an aqueous solution) with a salt content adjusted to allow the formation of particles. For example, a quarter saline solution is useful for this purpose. Due to the more efficient administration of the macrolide formulations provided by the present invention, much smaller volumes of macrolide are administered in a much shorter time than conventional administration regimens, thereby reducing administration costs and drug waste. And significantly increase the likelihood of patient compliance.
[0023]
Thus, according to one aspect of the present invention, there is provided a method for the treatment of a subject in need thereof (eg, a subject suffering from a susceptible endobronchial infection), the method comprising: Administering to the body a fixed dose spray aerosol formulation comprising from about 50 mg to about 750 mg of the macrolide and a pharmaceutically acceptable carrier. In one aspect of the invention, the aerosol formulation administered in the practice of the invention comprises from about 50 mg / ml to about 150 mg / ml macrolide antibiotic, preferably from about 70 mg / ml to about 130 mg / ml macrolide. It may be a liquid formulation comprising the antibiotic, and more preferably from about 90 mg / ml to about 110 mg / ml of the macrolide antibiotic. Preferably, a small volume aerosol formulation is administered to the subject. Thus, in this aspect, a dose of less than about 2.0 ml of the nebulized liquid aerosol formulation is administered to the subject. In another aspect, a dose of less than about 1.5 ml of a nebulized liquid aerosol formulation is administered to the subject. In yet another aspect, a dose of less than about 1.0 ml of a nebulized liquid aerosol formulation is administered to the subject.
[0024]
In another aspect, the macrolide compounds of the invention can be formulated for aerosol delivery as a dry powder. As used herein, the term "powder" refers to a free-flowing and easily dispersed in inhalation device that can then be inhaled by a subject, the particles of which reach the lungs and are located in the peripheral airways. A composition consisting of finely dispersed solid particles capable of infiltration and deposition. Thus, the powder formulations of the present invention are said to be "respirable." Preferably, the average powder particle size is less than about 10 μm in diameter and is a relatively uniform spheroid. More preferably, its diameter is less than about 7.5 μm, and most preferably about 5.0 μm. Usually, the particle size distribution is from about 0.1 μm to about 5 μm in diameter, especially from about 1 μm to about 5 μm. The dry powder formulation of the present invention has a moisture content such that the particles are easily dispersible in an inhalation device and form an aerosol. This water content is generally less than about 10% by weight (% w) water, usually less than about 5% w, and preferably less than about 3% w.
[0025]
The dry powder formulations of the invention generally comprise a therapeutically effective amount of a macrolide compound of the invention together with a pharmaceutically acceptable carrier. A dry powder formulation of the present invention may comprise from about 25 mg to about 250 mg of a macrolide antibiotic, preferably from about 50 mg to about 200 mg of a macrolide antibiotic, and more preferably from about 75 mg to about 150 mg of a macrolide antibiotic. . In this aspect of the invention, the dry powder formulation comprises from about 50% to about 90% by weight of the macrolide antibiotic, preferably from about 60% to about 88% by weight of the macrolide antibiotic, and more. Preferably about 75% to about 85% by weight of the macrolide antibiotic may be included.
[0026]
Suitable pharmaceutically acceptable carriers include those that can be taken into the lungs of a patient without significant harmful toxicological effects on the lungs, including, for example, stabilizers, Fillers, buffers, salts and the like. A sufficient amount of the pharmaceutically acceptable carrier is desired to ensure uniform pulmonary delivery to a subject in need thereof, with stability, dispersibility, consistency, and bulking properties. Used to get properties. The actual amount of pharmaceutically acceptable carrier used can be from about 0.05% w to about 99.95% w. More preferably, about 5% w to about 95% w of a pharmaceutically acceptable carrier is used. Most preferably, about 10% w to about 90% w of a pharmaceutically acceptable carrier is used.
[0027]
Pharmaceutical excipients useful as carriers in the present invention include stabilizers (human serum albumin (HSA)), fillers (saccharides, amino acids and polypeptides); pH adjusters or buffers; salts (eg, chlorides). Sodium) and the like. These carriers can be in crystalline or amorphous form or can be a mixture of the two. Preferred fillers include compatible carbohydrates, polypeptides, amino acids, or combinations thereof. Suitable carbohydrates include monosaccharides (galactose, D-mannose, sorbose, etc.); disaccharides (lactose, trehalose, etc.); cyclodextrins (eg, 2-hydroxypropyl-β-cyclodextrin; and polysaccharides (eg, Alditols such as mannitol, xylitol, etc. Preferred carbohydrate groups include lactose, trehalose, raffinose, maltodextrin, and mannitol Suitable polypeptides Amino acids include alanine and glycine, with glycine being preferred.Additives may be included as minor components of the dry powder formulations of the present invention, while steric during spray drying. For excipient stability and to improve the dispersibility of the powder, these additives may include hydrophobic amino acids such as tryptophan, tyrosine, leucine, phenylalanine, etc. Examples of the pH adjuster or buffer include organic salts prepared from an organic acid and a base (for example, sodium citrate, sodium ascorbate, etc.), and sodium citrate is preferred.
[0028]
In another aspect, the invention is directed to a concentrated macrolide formulation (eg, a concentrated erythromycylamine formulation) suitable for effective delivery of macrolide by aerosolization into the endobronchial space. The present invention relates to the treatment of Streptococcus pneumoniae infection, Haemophilus influenzae infection, Staphylococcus aureus infection, Moraxella catarrhalis infection, and Legionella pneumonia infection to Legionella pneumonia infection, Chlamynea pneumonia infection, and Legionella pneumonia infection. Suitable for the formulation of concentrated erythromycylamine for aerosolization with a jet nebulizer, vibrating porous plate nebulizer, ultrasonic nebulizer or dry powder nebulizer, which produces an erythromycilamine aerosol particle size of 1 μm to 5 μm. is there. The formulation is preferably tuned to allow for the production of erythromycylamine aerosol particles that are well tolerated by the patient but prevent the occurrence of undesired secondary side effects such as bronchospasm and cough. A minimal but still effective amount of erythromycylamine formulated in a relatively small volume of a physiologically acceptable solution with salt, or a dry powder.
[0029]
A key requirement for all aerosol formulations is their safety and efficacy. Further advantages are lower treatment costs, practicality of use, long shelf life, storage and nebulizer optimization.
[0030]
Its aerosol formulation is sprayed, the terminal and respiratory bronchioles (here, Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus and Moraxella catarrhalis, as well as atypical bacterium Legionella pneumonia, Chlamydia pneumoniae, and Mycoplasma pneumoniae or other Susceptible bacteria are present in patients with chronic bronchitis and pneumonia). Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus and Moraxella catarrhalis; However, they are most prevalent in terminal and respiratory bronchioles. During the hatred of infection, bacteria can also be present in the alveoli. Thus, in one aspect, the present invention provides a formulation that is delivered throughout the endobronchial tree from the tree to the terminal re-bronchi and ultimately to the parenchyma.
[0031]
Aerosolized erythromycylamine formulations are formulated for efficient delivery of erythromycylamine to the pulmonary bronchial lumen. Certain jet nebulizers, vibrating porous plate nebulizers or ultrasonic nebulizers use erythromycylamine aerosol particles (the median aerodynamic diameter of the size is significantly between 1-5 μm). Selected to allow the formation of Formulated, the amount of the delivered erythromycylamine is endobronchial infection (particularly bacterial Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus and Moraxella Catarralis and atypical pneumonia Legionella pneumonia, Chlamydia pneumoniae, and infection caused by Mycoplasma pneumoniae) of Effective for treatment and / or prevention. The formulation is salinized to allow the production of an erythromycylamine aerosol that is well tolerated by the patient. Further, the formulation has an appropriate osmolarity. This formulation has a small aerosolizable volume and can deliver an effective dose of erythromycilamine to the site of infection. In addition, the aerosolized formulation does not negatively impair airway function by producing unwanted side effects.
[0032]
The antibiotic formulation can be administered using an inhalation device that has a relatively high rate of aerosol burst, high firing dose efficiency, and firing limited by the patient during the actual inhalation period. Thus, while conventional air jet nebulizers exhibit aerosol ejection velocities on the order of 3 μl / sec, inhalation devices useful for use in the practice of the present invention typically have about 5 μl / sec or more, more preferably , About 6.5 μl / sec or more, and most preferably about 8 μl / sec or more. Further, conventional air jet nebulizers have relatively low propellant dose efficiencies, typically emitting only about 55% (or less) of the nominal dose as an aerosol, while being useful in the practice of the present invention. Useful inhalation devices typically release at least about 75%, more preferably at least about 80%, and most preferably at least about 85% of an aerosol-loaded dose for inhalation by a subject. In other aspects, conventional air-jet nebulizers typically deliver aerosolized drug throughout the delivery period, regardless of whether the subject is in the inhalation, exhalation, or stationary phases of the respiratory cycle. Release continuously, thereby wasting a substantial portion of the loading drug dose. In contrast, preferred inhalation devices for use in the practice of the present invention are actuated by respiration, limiting the delivery of aerosolized particles of the macrolide formulation to the actual duration of inhalation by the subject. An exemplary inhalation device that meets the above criteria and is suitable for use in the practice of the present invention is Aerodose ^TM Inhaler (commercially available from Aerogen, Inc., Sunnyvale, California). Aerodose ^TM Inhalers generate aerosol using a porous membrane driven by a piezoelectric oscillator. Aerosol delivery is activated by respiration and is restricted to the inhalation phase of the respiratory cycle (ie, aerosolization does not occur during the expiratory phase of the respiratory cycle). The airflow design allows for normal inhalation-expiratory breathing as compared to a breath-hold inhaler. In addition, Aerodose ^TM An inhaler is a portable, self-contained and easily transported inhaler. Piezoelectric oscillator aerosol generators (eg, Aerodose ^TM Inhalers, which are presently preferred for use in the practice of the present invention, meet the above performance criteria and can deliver small dosage volumes of the present invention with relatively high deposition rates within a relatively short period of time. An inhaler or nebulizer device may be used.
[0033]
In another aspect of the invention, unit dose formulations and devices provide for administration of a macrolide antibiotic formulation to a subject using an inhaler according to the methods of the invention described above. Preferred unit dosage devices are designed to hold and store a relatively small amount of the macrolide antibiotic formulation of the present invention and deliver the formulation to an inhalation device for delivery to a patient in aerosol form. Equipped with a container. In one aspect, the unit dose container of the present invention comprises a plastic ampule filled with a macrolide antibiotic formulation of the present invention and sealed under sterile conditions. Preferably, the unit dose ampoule is provided with a twist-off tab or other easily opened device for opening the ampoule and delivering the macrolide antibiotic formulation to the inhalation device. . Ampoules for containing drug formulations are well known to those of skill in the art (eg, US Pat. Nos. 5,409,125, 5,379,898, 5,213,860, and 5). Nos. 4,046,627, 4,995,519, 4,979,630, 4,951,822, 4,502,616 and 3,993,223. (See, these disclosures are incorporated herein by reference). The unit dose container of the present invention may be inserted directly into the inhalation device of the invention, and ultimately to the subject, to deliver the included macrolide antibiotic formulation. Can be designed.
[0034]
In accordance with this aspect of the invention, less than about 5.0 ml, preferably less than about 3.0 ml, and most preferably less than about 2.0 ml of liquid macrolide antibiotic formulation (in a physiologically acceptable carrier is less than about 5.0 ml). A unit dose device comprising a sealed container comprising 50 to about 150 mg / ml of a macrolide antibiotic is provided, wherein the sealed container is delivered to an inhalation device for aerosolizing the macrolide antibiotic formulation. Has been adapted to be. Suitable macrolide antibiotics for use in this aspect of the invention include those described in detail above. In a currently preferred embodiment, the macrolide antibiotic used in the unit dose device of the present invention is erythromycylamine. In another aspect of the invention, the unit dose device of the invention can include a liquid macrolide antibiotic formulation (including about 70 to about 130 mg / ml macrolide antibiotic). In still other aspects of the invention, a unit dose device of the invention can include a liquid macrolide antibiotic formulation (including about 90 to about 110 mg / ml macrolide antibiotic).
[0035]
In preferred liquid unit dose formulations of the present invention, the physiologically acceptable carrier allows for the generation of an erythromycylamine aerosol that is well tolerated by the patient, but with secondary undesired side effects such as bronchial A saline solution, such as a quarter strength normal saline solution, which is salt adjusted to substantially prevent the occurrence of convulsions and coughs).
[0036]
In yet another aspect of the invention, the dry powder formulation of the invention is suitable for providing a macrolide antibiotic compound of the invention to a subject in an amount sufficient for unit dosage treatment by dry powder inhalation. In a convenient unit dose storage container. Preferred dry powder unit dosage storage containers are those wherein the macrolide-based dry powder composition is aerosolized by dispersal in an air stream to form an aerosol, and the aerosol thus generated is then administered by a subject in need of treatment. Fitted in a suitable inhalation device that captures in a chamber fitted with a mouthpiece for inhalation of the mouthpiece. Such dosing storage containers include any container enclosing a formulation known in the art, such as a removable (eg, airflow) stream that directs the container to disperse the dry powder formulation. Gelatin capsules or plastic capsules having a portion). Such containers are exemplified by those shown in U.S. Pat. Nos. 4,227,522, 4,192,309, and 4,105,027. Suitable containers also include those used with Glaxo's Ventolin Rotohaler brand powder inhaler or Fison's Spinhaler brand powder inhaler. Another suitable unit dose container that provides an excellent moisture barrier is formed from aluminum foil plastic sheet. The macrolide powder is filled by weight or volume at low pressure into a formable foil and sealed with a foil-plastic wrap. Such a container for use with a powder inhalation device is described in U.S. Patent No. 4,778,054 and disclosed in Glaxo's Diskhaler. RTM. (U.S. Pat. Nos. 4,627,432, 4,811,731; and 5,035,237). All of these references are incorporated herein by reference.
[0037]
According to this aspect of the invention, the dry powder formulation (from about 25 to about 250 mg of a macrolide antibiotic in a physiologically acceptable dry powder carrier, preferably from about 50 to about 200 mg of a macrolide antibiotic) , More preferably, comprising a sealed container comprising about 75 to about 150 mg of a macrolide antibiotic, the sealed container being provided to the inhalation device for aerosolization. It is adapted to deliver a formulation. In this aspect of the invention, the dry powder formulation comprises from about 50% to about 90% by weight of a macrolide antibiotic, preferably from about 60% to about 88% by weight of a macrolide antibiotic, Preferably, it may contain from about 75% to about 85% by weight of a macrolide antibiotic.
[0038]
(Aerosol erythromycylamine formulation)
To evaluate the stability of erythromycylamine in aqueous solution, three salt forms of this antibiotic were prepared and subjected to various conditions of temperature, time, concentration and pH. Erythromycylamine concentration was determined by HPLC methodology. The data from these stability studies are shown in FIGS. 2-7 and reveal some important findings. First, the stability of erythromycylamine hydrochloride was, as expected, directly proportional to the temperature of the solution (see FIGS. 2-5). Second, the erythromycylamine solution was more stable at neutral pH 7 than at acidic pH 5 and 6 (FIG. 5). This result is consistent with the known effect of pH on macrolide antibiotic degradation. One of the major degradation pathways is the loss of the neutral sugar, cladinose (see J. Chrom. A, 812m 1998, 255-286). Third, solutions of erythromycylamine acetate were more stable at the same pH than the corresponding hydrochlorides and sulfates at pH 6 and 7 (compare FIG. 7 with FIGS. 5 and 6).
[0039]
Liquid and dry powder formulations according to the present invention comprise from about 50 to about 750 mg, preferably from about 75 to about 600 mg, most preferably from about 100 to about 500 mg of macrolide antibiotic drug per dose (e.g., Erythromysylamine acetate). These include Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Moraxella catarrhalis, Legionella pneumonia, and Chlamydia pneumoniae.
[0040]
A presently preferred liquid aerosol erythromycylamine formulation according to the present invention comprises about 90 to about 110 mg erythromycylamine sulphate per ml of normal saline at a quarter concentration. This represents a representative effective amount of erythromycylamine for controlling bacterial infection of AECB.
[0041]
Both patients and aerosol generating devices are sensitive to the osmotic pressure, pH, and ionic strength of the formulation. The problem is that a quarter concentration of normal saline (this is a saline solution containing 0.225% sodium chloride, and this quarter concentration of normal saline is injected into the bronchial lumen It has now been found that this is solved conventionally by formulating the erythromycylamine solution in erythromycylamine (which is a suitable vehicle for delivery of erythromycylamine).
[0042]
Chronic bronchitis patients and other patients with chronic intrabronchial infection have a high incidence of bronchospasm or asthmatic airways. These airways are sensitive to hypotonic or hypertonic aerosols, concentrations of permeable ions (especially halides such as chloride), and acidic or basic aerosols. The effect of irritating the respiratory tract can be manifested clinically by coughing or bronchospasm. Both of these conditions can prevent efficient delivery of aerosolized erythromycylamine to the bronchial lumen.
[0043]
Formulations of erythromycylamine acetate, erythromycylamine hydrochloride and erythromycylamine sulfate (containing 60-100 mg of erythromycylamine / ml of normal saline at quarter concentration) have a range of 130-400 mOsm / kg. Has osmotic pressure. This is within the safe range of aerosols administered to patients with chronic bronchitis (Table 1).
[0044]
[Table 1]

The pH of this formulation is equally important for aerosol delivery. As previously noted, if the aerosol is either acidic or basic, it can cause bronchospasm and cough. The safety range of pH is relative; some patients tolerate mildly acidic aerosols that cause bronchospasm in others. Any aerosol with a pH below 4.5 usually induces bronchospasm in hypersensitive individuals; aerosols with a pH between 4.5 and 5.0 occasionally cause this problem. Aerosols having a pH between 5.0 and 7.0 are considered safe. Any aerosol with a pH greater than 10.0 should be avoided. This is because a stimulus that causes bronchospasm can occur. The optimal pH for the aerosol formulation was determined to be between pH 5.0 and 7.0.
[0045]
In one aspect, the liquid formulations of the present invention are preferably nebulized to a particle size that allows for the delivery of the drug primarily to the terminal and respiratory bronchioles where bacteria are present, as well as to the lower airways. . Efficient delivery of erythromycilamine to the pulmonary bronchial airways by aerosol requires a formulation of aerosol particles having a median aerodynamic diameter of primarily between 1-5 μm. Prevention of endobronchial infections (especially for Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Moraxella catarrhalis, Legionella pneumonia, Chlamynea, Chlamynea and Chlamyamine infections) The amount given must efficiently target the inner bronchial surface. The delivered dose of the formulation preferably has a particle aerosolizable volume that is minimal to deliver an effective dose of erythromycylamine to the site of infection. Preferred formulations further provide conditions that do not adversely affect airway function. As a result, preferred formulations include a sufficient amount of the drug formulated under conditions that allow for efficient delivery of the drug, while avoiding undesired reactions. The new formulations according to the invention fulfill all of these needs.
[0046]
In accordance with the present invention, erythromycylamine is formulated into a dosage form intended for inhalation therapy by patients with chronic bronchitis and pneumonia. Since patients are present worldwide, it is desirable that this formulation have a reasonably long shelf life. Therefore, storage conditions and formulation stability are important.
[0047]
As discussed above, the pH of this solution is important. In terms of storage and longer shelf life, a pH between 5.0 and 7.0, preferably about 6.0, is optimal.
[0048]
The formulation is typically stored in a 1-2 milliliter low density polyethylene (LDPE) vial. The vial is aseptically filled using a blow-fill-seal process. The vial is sealed with a foil overpouch.
[0049]
The stability of this formulation with respect to oxidation is another very important issue. If the drug is broken down prior to aerosolization, less drug will be delivered to the lungs, and thus the delivered dose will be very small, impeding treatment and reducing resistance to erythromycilamine. Trigger conditions that can lead to expression. In addition, erythromycylamine degradation products can induce bronchospasm and cough. Packaging the LDPE vial in an oxygen protective package containing a foil overpouch (6 vials per overpouch) to prevent oxidative degradation of erythromysylamine and to provide acceptable stability By doing so, a product with a low oxygen content is produced. Prior to vial filling, the solution in the mixing tank is sparged with nitrogen and the annular overpouch headspace is purged with nitrogen. In this way, both hydrolysis and oxidation of erythromysylamine are prevented.
[0050]
(II. Aerosolization device)
Aerosolization devices useful in the practice of the present invention (eg, jet nebulizers, vibrating porous (vibrating porous) plate nebulizers, or ultrasonic nebulizers) generally provide formulations of the present invention primarily in the 1-5 μm range. Can be sprayed on aerosol particles. By predominantly in the present application is meant that at least 70%, but preferably more than 90%, of all aerosol particles produced are in the range of 1-5 μm.
[0051]
Nebulizers (e.g., jet nebulizers, ultrasonic nebulizers, vibrating porous (vibrating porous) plate nebulizers), and high voltage dry powder inhalers (Streptococcus pneumoniae, Haemophilus influenzae, Staphylocium aureus, Moraxella ichalai latina rialica, laurac, uraicella, italy, italy) Suitable for the treatment of Mycoplasma pneumoniae infection, which can produce and deliver particles between particle sizes between 1 and 5 μm) are currently available or can be made using known methods and materials . Jet nebulizers work by air pressure to break liquid solutions into aerosol droplets. Vibrating porous (vibrating porous) plate nebulizers work by using sonic vacuum generated by a rapidly vibrating porous plate to push solvent droplets through the porous plate. Ultrasonic nebulizers work by means of a piezoelectric crystal that splits a liquid into small aerosol droplets. However, only some formulations of erythromycylamine can be effectively nebulized by these three nebulizers. This is because these devices are sensitive to the pH and ionic strength of the formulation.
[0052]
Although various devices are available, only a limited number of these atomizers are suitable for the purposes of the present invention. Preferred nebulizers useful in the present invention include, for example, AeroNeb ^TM And AeroDose ^TM Vibrating porous plate nebulizer (AeroGen, Inc., Sunnyvale, Galif.), Sidestream® nebulizer (Medic-Aid Ltd., West Sussex, England), Pari LC Plus® and Pari LC® Trade Mark Jet nebulizer (Pari Respiratory Equipment, Inc., Richmond, Virginia), and Aerosonic ^TM (DeVilbiss Medizinische Produkte (Deutschland) GmbH, Heiden, Germany) and UltraAire® (Omron Healthcare, Inc., Vernon Hills, Illinois).
[0053]
(III. Pharmacokinetics of aerosol)
Solutions of erythromycylamine were administered to rats via the IV and inhalation routes, and plasma and lung drug concentrations were measured. Data from these studies are shown in FIGS. Two dose levels, 1.7 mg / kg and 0.7 mg / kg, were selected for the inhalation delivery route and compared against a single intravenous dose (25 mg / kg).
[0054]
Dose normalized AUC of erythromycilamine in the lung for IV (25 mg / kg), inhalation (1.7 mg / kg), and (0.7 mg / kg) are 24.21 μg · h / gram, 1067. It was 84 µg · h / gram and 848.34 µg · h / gram. Thus, by administering erythromycilamine directly to the lung via the inhalation route, the pulmonary drug levels achieved were approximately 40-fold higher on a milligram basis than delivery by the intravenous route. Thus, antibiotic therapy by inhalation should be more effective than treatment by the oral or IV route.
[0055]
(IV. Effectiveness of aerosol)
Erythromycylamine was very effective for both intravenous and aerosol administration. At the lowest dose tested (10 mg / kg per day), intravenous erythromycylamine, as shown in Example 7, The pneumonia lung load was reduced below the limit of detection (10 CFU / grams of lung). Aerosols are also very effective (see FIG. 10) with 5 mg / ml aerosol solution (calculated dose 0.13 mg / kg per day). With only detectable recovery of pneumonia. In addition, erythromycylamine was very effective when administered as a single aerosol dose at a concentration greater than that required for a single daily dose for 3 days. A single dose of 0.13 mg / kg was less effective (less than 2 digits as CFU / gram) compared to 0.13 mg / kg (5 orders of magnitude reduction) for 3 consecutive days. Decrease). However, with a single dose of 0.67 mg / kg, almost complete clearance of the organism from lung tissue was achieved. This effect is similar to multiple dose efficacy, indicating that at this concentration the second and third doses have little value (see FIG. 11).
[0056]
Pharmacokinetic evaluation of aerosolized erythromycylamine suggests, and efficacy data suggests that lung concentrations equivalent to multiple daily IV, oral, or aerosol doses were What can be achieved with a single aerosol dose and that this single dose is about 3-5 times greater than required for similar efficacy as three daily doses.
[0057]
(Usefulness)
One aspect of the utility of the present invention is that low volume, high concentration formulations of macrolide antibiotics (eg, erythromysilamine) can be used with a suitable nebulizer to provide chronic bronchitis, bronchiectasis, And delivering an effective dose of erythromycilamine to the endobronchial space of a person with pneumonia (caused by macrolide-sensitive bacteria or other infections). This formulation is safe and very cost effective. Further, the formulation can be kept under a nitrogen environment with a pH controlled for resistance to provide a suitable shelf life for commercial distribution.
[0058]
(Example 1)
General Procedure for Preparation of Erythromycylamine Salt: Synthesis of Erythromycylamine Acetate
To a solution of 10.0 g (13.6 mmol) of erythromycylamine in 100 mL of MeOH (cooled in an ice bath) was added 1.56 mL (27.2 mmol, 2.0 equiv) of glacial acetic acid dropwise. added. The solution was warmed to ambient temperature over 30 minutes and then the solvent was removed under reduced pressure. Et ₂ O (50 mL) was added and the slurry was concentrated. This was repeated to give 11.52 g (96.9%) of erythromycylamine monohydrate acetate as a white powder; IR (KBr, cm ^-1 ) 1718, 1560, 1406, 1168, 1080, 1055, 1012; ¹ HNMR (400 MHz, CD ₃ OD) [delta] 0.89 (t, 3H, J = 7.2 Hz), 1.06-1.32 (m, 27H), 1.35-1.47 (m, 4H), 1.52-1.66. (M, 3H), 1.85-2.02 (m, 8H), 2.03-2.26 (m, 2H), 2.45-2.49 (m, 1H), 2.66-2 .77 (m, 5H), 2.91-3.09 (m, 3H), 3.21-3.40 (m, 6H), 3.58 (d, 1H, J = 7.0 Hz), 3 .67 (s, 1H), 3.78-3.83 (m, 2H), 4.10-4.13 (m, 1H), 4.59 (d, 1H, J = 7.0 Hz), 4 .88-5.01 (m, 12H); MS m / z 735.6 (M ⁺ -2AcOH-2H ₂ O); KF 2.33% H ₂ O.
[0059]
C ₄₁ H ₈₀ N ₂ O ₁₇ Calculated for: C, 56.40; H, 9.24; N, 3.21. Found: C, 56.38; H, 9.21; N, 3.16.
[0060]
(Example 2)
(Synthesis of erythromycylamine sulfate)
0.73 mL (13.6 mmol, 1.0 equivalent) of concentrated sulfuric acid was added dropwise to a solution of 10.0 mL (13.6 mmol) of erythromycylamine in 100 mL of MeOH (cooled in an ice bath). added. The solution was warmed to ambient temperature over 30 minutes and then the solvent was removed under reduced pressure. Et ₂ O (50 mL) was added and the slurry was concentrated. This was repeated to give 11.13 g (96.1%) of erythromysylamine monohydrate sulfate as a white powder; IR (KBr, cm ^-1 ) 1718, 1384, 1168, 1122, 1078, 1012; ¹ HNMR (400 MHz, CD ₃ OD) [delta] 0.89 (t, 3H, J = 7.2 Hz), 1.08-1.32 (m, 27H), 1.45-1.63 (m, 7H), 1.89-2.04 (M, 2H), 2.23-2.31 (m, 2H), 2.44-2.47 (m, 1H), 2.84-2.89 (5H), 2.99-3.07 (M, 3H), 3.30-3.49 (m, 6H), 3.58 (d, 1H, J = 7.0 Hz), 3.69 (s, 1H), 3.78-3.86 (M, 2H), 4.09-4.11 (m, 1H), 4.60 (d, 1H, J = 6.8 Hz), 4.87-4.99 (m, 12H); MS m / z 735.7 (M ⁺ -H ₂ SO ₄ -2H ₂ O); KF 2.93% H ₂ O.
[0061]
C ₃₇ H ₇₄ N ₂ O ₁₆ Calculated value for S: C, 52.22; H, 8.76; N, 3.29. Found: C, 52.55; H, 8.91; N, 3.27.
[0062]
(Example 3)
(Synthesis of erythromysylamine hydrochloride)
2.34 mL (27.2 mmol, 2.0 eq) of 37% hydrochloric acid was added dropwise to a solution of 10.0 g (13.6 mmol) of erythromyclamine in 100 mL of MeOH cooled in an ice bath. The solution was allowed to warm to ambient temperature over 30 minutes and then the solvent was removed under reduced pressure. Et ₂ O (50 mL) was added and the slurry was concentrated. This was repeated until obtaining 11.24 g (97.9%) of erythromycylamine hydrochloride dihydrate as a white powder; IR (KBr, cm ^-1 ) 1718, 1466, 1383, 1170, 1078, 1055, 1011; ¹ 1 H NMR (400 MHz, CD ₃ OD) [delta] 0.87-0.91 (m, 3H), 1.10-1.31 (m, 27H), 1.43-1.65 (m, 7H), 1.89-2.01 (m , 2H), 2.25-2.27 (m, 2H), 2.45-2.48 (m, 1H), 2.82-3.10 (m, 8H), 3.34-3.42. (M, 8H), 3.57-3.58 (m, 1H), 3.67 (s, 1H), 3.80-3.82 (m, 2H), 4.08-4.11 (m , 1H), 4.61-5.00 (m, 13H); MS m / z 735.6 (M ⁺ -2HCl-2H ₂ O); KF 4.38% H ₂ O.
[0063]
analysis. C ₃₇ H ₇₆ Cl ₂ N ₂ O ₁₂ For C: 52.65; H, 9.08; N, 3.32. Found: C, 52.21; H, 9.18; N, 3.20.
[0064]
(Example 4)
(Stability of Aqueous Formulation and Erythromycylamine Salt)
(Preparation of Solution) Erythromycylamine (9.0 g, 12.2 mM) free base was added to a tared 100 mL Erlenneyer flask. Deionized water (25 mL) was added to the flask with magnetic stirring. 1N sulfuric acid (24.5 mL, 2 eq) was added slowly with stirring. When the solution became clear, the flask was removed from the stir plate and reweighed. Deionized water was added dropwise to give a final solution weight of 62.9 g. The solution is divided into three 20 mL portions and the pH is monitored by a pH meter and the desired value (5.0, 6.0, or 7.0) is added by dropwise addition of 1 N sodium hydroxide or sulfuric acid. Adjusted to 0). Using the above procedure, adjust the weight of erythromysylamine (6.0 g and 3.6 g) and the volume of 1 N sulfuric acid (16.3 mL and 9.8 mL) to give 100 mg / mL and 60 mg / mL. Was prepared.
[0065]
A solution of 150, 100 and 60 mg / mL erythromycylamine acetate and hydrochloride was prepared by adding 1N acetic acid and 1N hydrochloric acid (2 equivalents) to prepare the salt and adjust the pH. Prepared as described above.
[0066]
Aliquots of each salt form at each concentration and each pH were stored at 4, 40, and 60 ° C, and at ambient temperature.
[0067]
(Determination of Stability) All solutions were analyzed immediately after preparation (t = 0), 24 hours, 48 hours, 8 days, 15 days, and 22 days after preparation, except that on day 8, Samples that were degraded were excluded from further analysis).
[0068]
Refrigerated and heated samples were allowed to equilibrate to ambient temperature for at least one hour prior to sample preparation. The final dilution volume for all samples was 10 mL. The diluent for all samples consisted of a 80:20 (v / v) mixture of 50 mM phosphate buffer (pH 6.5) and acetonitrile.
[0069]
The appropriate amount of sample (40 microliters for a 150 mg / mL solution, 50 microliters for a 100 mg / mL solution, or 100 microliters for a 60 mg / mL solution) was transferred to a 20 mL scintillation vial. 10 mL of diluent was added to the vial and mixed thoroughly.
[0070]
Preparation of Standards Standards were prepared in duplicate and used for up to 3 days. Ery-amine free base (30 mg) was transferred to a tared 50 mL volumetric flask and the exact weight recorded. Sample diluent (45 mL) was added and briefly sonicated until dissolved. The standard was cooled and diluted to volume with diluent.
[0071]
(Analysis of Samples and Standards) Samples and standards were analyzed by reversed-phase high-performance liquid chromatography. Separation was performed using a 250 x 4.6 mm Phenomenex Luna CN column with a 5 micron particle size. All analyzes were performed on an Agilent Technologies HP 1100 chromatography system, and data was collected and recorded using an Agilent Technologies ChemStation data system. The analysis parameters were as shown in Table 2 below.
[0072]
(Table 2)
Flow rate 1.0mL / min
Column temperature 30 ° C
Injection volume 20μL
Detector UV absorbance (200nm)
Execution time 10 minutes
Mobile phase A 50 mM phosphate pH 2.1
Mobile phase B acetonitrile
Composition 80/20 A / B.
[0073]
(Example 5)
(Osmolality of erythromycylamine salt solution)
Three portions of erythromycylamine HCl salt (0.6 g, 1.0 g and 1.5 g) were weighed into separate 10 mL volumetric flasks. Easypure UV water (8 mL) was added to each flask and sonicated until completely dissolved, then diluted to volume. This procedure was repeated for erythromycylamine sulfate and acetate. The pH and osmolality of each solution was measured, and the measured osmolality was compared to the theoretical value.
[0074]
The salt prepared in Example 4 (4 ° C.) was equilibrated to room temperature and the osmolality was measured. Table 3 shows the results.
[0075]
(Table 3)
[0076]
[Table 2]

(Example 6)
(Aerosol delivery of erythromycylamine to rats :)
(Characterization of aerosol pharmacokinetics)
(Intravenous pharmacokinetics): Erythromycylamine (250 mg) was dissolved in 5 mL of DI water and 12 mL of concentrated sulfuric acid was added. A solution of dilute sulfuric acid (1:10 v / v) was gradually added to this solution to completely dissolve the drug. A solution of dilute sulfuric acid was slowly added to bring the pH of the solution to 6.8-7.2. The total volume of the solution was brought to 8 mL by the addition of DI water. 200 μl (25 mg / kg) of a solution of erythromycylamine sulfate was delivered to male Sprague-Dawley rats (Simonsen Laboratories, 1180 C Day Road, Gilroy, CA 95020) by intravenous administration via the lateral tail vein. Animals were anesthetized with 1-4% isoflurane and lung and blood samples were collected from three rats at doses of 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and Collected after 24 hours. Blood samples were collected via cardiac puncture using heparin as an anticoagulant. Lungs were surgically removed after blood sampling and the bronchi and trachea were removed and discarded. The remaining lung tissue was processed as follows. Both lung and blood samples were immediately placed on ice and blood samples were centrifuged immediately after collection to collect plasma samples. Both lung and plasma samples were stored at -80 C until assayed.
[0077]
Plasma and lung (per g lung tissue) concentrations of erythromycylamine were determined using validated LC-MS methods. Plasma samples (100 μg) were spiked with oleandmycin (internal standard, 1 μg / mL) prior to extraction. Plasma samples (100 μL) were deproteinized using 3.3% trichloroacetic acid (TCA). The sample was centrifuged (10,000 rpm, 10 minutes) and the supernatant was transferred to an HPLC centrifugal filter (10,000 rpm, 10 minutes) for centrifugation. The mobile phase was a 0.1% acetic acid-acetonitrile (70:30, v / v, pH = 3.2) solution (3 min at a flow rate of 0.5 ml / min), followed by a 0.1% acetic acid-acetonitrile ( 60:40, v / v) (3 min at a flow rate of 0.8 ml / min). A stainless steel analytical column (Zorbax SB-C18, 2.1 mm ID x 150.0 mm, 5 μm and Phenomenex cartridge guard column) was used as the stationary phase. The column temperature was 50 ° C. Quantification of erythromycylamine was performed using an HP 1100 LC / MSD API Electrospray System. Data acquisition was set to the selective ion monitoring mode. This method was linear (r> 0.9990) in the concentration range of 0.01-50 μg / mL. Absolute recovery was 95.0 ± 2.19%.
[0078]
Lung samples were homogenized with DI water. Oleandomycin was added to this sample as an internal standard. The protein was removed from this homogenate using 0.9 M TCA. The sample was centrifuged at 10,000 rpm for 10 minutes and the supernatant was transferred to an HPLC centrifugal filter for centrifugal filtration. The mobile phase was 0.1% acetic acid-acetonitrile (70:30, v / v, pH = 3.2) (3 min at a flow rate of 0.5 ml / min), followed by 0.1% acetic acid-acetonitrile (60%). : 40, v / v) (3 min at a flow rate of 0.8 ml / min). A stainless steel analytical column (Zorbax SB-C18, 2.1 mm ID x 150.0 mm, 5 μm and Phenomenex cartridge guard column) was used as the stationary phase. Column temperature was 50 ° C. Quantification of erythromysylamine was performed using an HP 1100 LC / MSD API-Electrospray System. Data acquisition was set to the selective ion monitoring mode. The linearity of this assay (r> 0.9990) ranged from 0.1 to 200 μg / g. The extraction efficiency was 93.8 ± 2.54%.
[0079]
Pharmacokinetic parameters, area under the curve (AUC) and mean residence time (MRT) were determined using WinNonlin ^TM Estimation was based on statistical moment theory using Professional Version 2.0 software (Pharsight Corporation). Peak concentration (C _max ) Were not estimated, but were observed.
[0080]
(Pharmacokinetics of inhalation): For a solution of 60 mg / mL, 3.191 g (4.08 mmol) of erythromycin was added to 43 mL of DI water and 4.27 mL (4.27 mmol) of 1 M sulfuric acid in a 50 ml volumetric flask. Luamine (94% purity) was added. The solution was then adjusted to pH 6.5 by adding an additional 53 μL (0.053 mmol) of 1 M sulfuric acid. The volume was brought to 50 mL with additional DI water. A 30 mg / mL solution was made by diluting the 60 mg / mL solution with normal saline to 1/2. The osmolality of the resulting solution was measured using The Advanced ^TM It was 148 mOsm as determined using a Micro-Osmometer model 3300 (Advanced Instruments, Inc., Norwood, Mass.).
[0081]
Rats are treated with 30 mg / mL of erythromycilamine amine sulphate for 30 minutes via inhalation in a 32-port no-only rodent exposure system (Battelle, Richland, WA) with a 32 port nose-only rodent exposure system. Or one exposure to either the 60 mg / mL solution. The Battellle system nose-only rodent exposure system is based on the Cannon Flow-Past Nose only system (Am. Ind Hyg Assoc J 1983 Dec; 44 (12) 923-8) and has a total of 32 ports and 4 ports. Composed of stackable stainless steel layers. The system provided monitoring and control of inlet and outlet streams, and aerosol data was collected using NORES version 1.1.4 software provided by Battell. Transfer the erythromycylamine solution to PARI LC STAR ^TM Aerosolized using a nebulizer. Average aerosol concentrations were determined by gravimetric analysis of filter samples taken 10 and 20 minutes after the start of exposure. Average aerosol concentrations were 0.54 ± 0.06 and 1.36 ± 0.30 mg / L for the 30 mg / mL and 60 mg / mL solutions, respectively.
[0082]
Lung and blood samples were collected from three rats at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after dosing as described above. Sample collection and handling procedures for this inhalation study were similar to those for the intravenous study.
[0083]
The procedure of the bioanalytical assay for inhalation studies was similar to that for intravenous studies. The calculated deposited dose in the lung (pulmonary dose) was approximately 0.70 mg / kg or 1.77 mg / kg, respectively, after a 30 minute inhalation dose of a 30 mg / mL or 60 mg / mL erythromycilamine solution. . The lung dose in the lung was calculated as follows:
LDD = MAC × MV × DE × FLD @ MBW
here,
LDD = dose deposited in lung
MAC = average aerosol concentration = 0.54 and 1.36 mg / L for solutions of 30 and 60 mg / mL, respectively.
MV = ventilation volume per minute = 0.1 L / min
DE = duration of exposure = 30 minutes
FLD = pulmonary deposition fraction = 0.1
MBW = average weight = 0.23 kg.
[0084]
The pharmacokinetic parameters in the lung after intravenous and inhaled administration of erythromycylamine are summarized in Table 4 below:
(Table 4)
(Pharmacokinetic parameters of erythromycilamine in the lung after intravenous dosing or two inhaled doses in rats (N = 3))
[0085]
[Table 3]

1. Area under the curve estimated from 0 to 24 hours after dosing
Area under the curve normalized to 2.1 mg / kg
3. Average residence time estimated from 0 to 24 hours after dosing
n. e. : Not estimated.
[0086]
(Example 7)
Aerosol and Intravenous (IV) Efficacy of Erythromycylamine in S. Pneumonia Rat Lung Model of Infection
(Method)
Male Sprague-Dawley rats were cultured with 50-100 microliters of S. aureus prepared in agar beads. Infection was achieved by intratracheal administration of Pneumonia A66 (strain number PGO4716). Prepared by suspending the inoculum in a broth culture of PGO4716 in molten agar and suspending the agar suspension with mixing in sterile mineral oil to produce small beads of agar containing the bacteria. did. The beads were collected by centrifugation, resuspended in sterile saline, and administered to each animal by direct injection into the lung via tracheostomy.
[0087]
An erythromycylamine solution is prepared in sterile saline. Antibiotics were administered either by intravenous injection into the tail vein or by aerosol exposure. This aerosol exposure was achieved by nasal only exposure using the In-Tox Aerosol Exposure System (Model No. 04-1100). The system includes a rodent secured to a plastic tube that is open to the system at one end (nose port) and sealed at the other end to maintain system integrity. A closed aerosol delivery system designed for exposure. The aerosol is supplied at a flow rate of about 6.5 liters / minute at Pari LC Star. ^TM Produced by an air jet nebulizer. The suction is set at 9 l / min, so that the total flow through this system with leaner air is 7.5 l / min.
[0088]
Treatment begins 24 hours after infection and lasts 3 days, once a day. Aerosols were administered daily for 30 minutes. On day 4 post infection and 12 hours after the last dose, the animals were sacrificed and the lungs were surgically removed. After this resection, the lungs were homogenized, diluted and quantitatively plated on blood agar. The plate was incubated for 24 hours and Pneumoniae colonies are counted to determine bacterial uptake. The results are shown in Table 5:
(Table 5)
(Efficacy of erythromycylamine against S. pneumoniae in rat Pneumoniae model)
[0089]
[Table 4]

* BQL = below the limit of quantification
(Example 8)
(Efficacy of aerosol of erythromycylamine in a S. Pneumonia rat lung model of infection after a single dose treatment of erythromycylamine)
Male Sprague-Dawley rats were infected and exposed to aerosol treatment as described in Example 7. This single treatment was initiated 24 hours after infection with an aerosol administered at the indicated dose over 30 minutes. No further treatment was performed and the animals were observed until surgery. On day 4 post infection (day 3 post dosing), the animals were sacrificed and their lungs removed surgically. After this resection, the lungs were homogenized, diluted and quantitatively plated on blood agar. The plate was incubated for 24 hours and Pneumoniae colonies are counted and bacterial uptake is determined. The results after a single dose administration are shown in FIG. Further results are shown in Table 6:
(Table 6)
(Efficacy of erythromycylamine against S. pneumoniae in rat Pneumoniae model)
[0090]
[Table 5]

BQL = below the limit of quantification.
[0091]
(Example 9)
(Aerosol delivery of erythromycylamine to dogs :)
(Characterization of aerosol pharmacokinetics)
(Pharmacokinetics of inhalation :)
For a solution of 60 mg / mL, to 43 mL of DI water and 4.27 mL (4.27 mmol) of 1 M sulfuric acid in a 50 ml volumetric flask, add 3.191 g (4.08 mmol) of erythromycylamine (94% purity). did. The solution was then adjusted to pH 6.5 by adding an additional 53 μL (0.053 mmol) of 1 M sulfuric acid. The volume was brought to 50 mL with additional DI water. Dogs were exposed once to a 60 mg / mL solution of erythromysylamine sulfate via an inhalation mask exposure system (Inversk Research, Scotland, UK) for 30 minutes.
[0092]
These dogs were removed from their enclosure in the dog holding area and transferred to the dosing lab. During dosing, the animals were restrained either by animal attendants or with a hanging / harness system. Inhalation dosing was performed using a sealed face mask connected to a properly characterized nebulizer prior to the start of dosing. The dosing device incorporated a face mask and mouthpiece attached to a flexible tube, which was connected to a nebulizer device. The mouthpiece was positioned on the tongue in the mouth of the animals, and the face mask was sealed around the dog nose with a rubber sleeve. The exhaust valve from this mask was connected to an exhaust system. When the dosing device is fully assembled and adapted to the dog, inspiration is indicated by the movement of the aerosol to the dog through a flexible tube.
[0093]
Lung samples were collected from two dogs at 2, 24, 48, 72, 96, and 120 hours after dosing. The lungs were surgically resected from this dog and the lobes (right tail (lower right lobe), left tail (lower left lobe), right head (upper right lobe), left head (upper left lobe), right center (right The middle lobe) and the accessory lobe) were separated for the assay. Plasma samples were collected from all surviving animals at 2, 24, 48, 72, 96, and 120 hours.
[0094]
The concentration of erythromycylamine in plasma and lung (per gram of lung tissue) was determined using LC-MS method. Plasma samples (100 μg) were spiked with oleandmycin (internal standard, 1 μg / mL) prior to extraction. Plasma samples (100 μL) were deproteinized using 3.3% trichloroacetic acid (TCA). The sample was centrifuged (10,000 rpm, 10 minutes) and the supernatant was transferred to an HPLC centrifugal filter (10,000 rpm, 10 minutes) for centrifugation. The mobile phase was 0.1% acetic acid-acetonitrile (70:30, v / v, pH = 3.2) solution (3 min at a flow rate of 0.5 ml / min) followed by 0.1% acetic acid-acetonitrile (60%). : 40, v / v) (3 min at a flow rate of 0.8 ml / min). A stainless steel analytical column (Zorbax SB-C18, 2.1 mm ID x 150.0 mm, 5 [mu] m and Phenomenex cartridge guard column) was used as the stationary phase. The column temperature was 50 ° C. Quantification of erythromycylamine was performed using an HP 1100 LC / MSD API Electrospray System. Data acquisition was set to the selective ion monitoring mode. This method was linear (r> 0.9990) in the concentration range of 0.01-50 μg / ml. Absolute recovery exceeded 90%.
[0095]
Lung samples were homogenized with DI water. Oleandomycin was added to this sample as an internal standard. Protein was removed from this homogenate using 0.9M TCA. The sample was centrifuged at 10,000 rpm for 10 minutes and the supernatant was transferred to an HPLC centrifugal filter for centrifugal filtration. The mobile phase was 0.1% acetic acid-acetonitrile (70:30, v / v, pH = 3.2) (3 min at a flow rate of 0.5 ml / min) followed by 0.1% acetic acid-acetonitrile (60:60). 40, v / v) (3 min at a flow rate of 0.8 ml / min). A stainless steel analytical column (Zorbax SB-C18, 2.1 mm ID x 150.0 mm, 5 [mu] m and Phenomenex cartridge guard column) was used as the stationary phase. Column temperature was 50 ° C. Quantification of erythromysylamine was performed using an HP 1100 LC / MSD API-Electrospray System. Data acquisition was set to the selective ion monitoring mode. The linearity of this assay (r> 0.99) ranged from 2 to 100 μg / g for lung. The extraction efficiency was over 90%.
[0096]
Pharmacokinetic parameters, area under the curve (AUC) and mean residence time (MRT) and half-life (T _1/2 ) To WinNonlin ^TM Estimation was based on statistical moment theory using Professional Version 3.1 software (Pharsight Corporation). Peak concentration (C _max ) Were not estimated, but were observed.
[0097]
Pharmacokinetic parameters in pulmonary inhalation and plasma inhalation following administration of erythromycilamine in dogs are summarized in Table 7 below and in FIGS. 13 and 14:
(Table 7)
(Pharmacokinetic parameters of erythromycilamine in lung and plasma after 30 min inhalation administration of a 60 mg / mL solution in dogs (N = 2))
[0098]
[Table 6]

1. Average residence time
n. e. : Not estimated.
[0099]
(Example 10)
(Liquid aerosol delivery of erythromycylamine)
A solution of erythromycylamine sulfate (100 mg / mL) in 1/4 (quater) normal saline (pH 7.0) is prepared according to the general procedure of the preceding example. A 1.0 mL dose of this solution is administered to a human subject suffering from chronic bronchitis (AECB) in acute exacerbation phase, AeroGen Aerodose. ^TM Administer by aerosol inhalation in less than 10 minutes using an inhaler. A reduction in AECB and the bacteria involved in the symptoms of AECB is observed.
[0100]
(Example 10)
(Dry powder aerosol delivery of erythromycylamine)
A dry powder formulation of erythromycylamine sulfate (100 mg) and a dry powder carrier (equal parts lactose, 2-hydroxypropyl-β-cyclodextrin, mannitol, and aspartame; total mass 25 mg) are prepared. The formulation is administered to a human subject suffering from chronic bronchitis (AECB) in the acute exacerbation period, and Glaxo Ventolin Rotohale. ^TM Administer by aerosol inhalation for less than 2 minutes using an inhaler. A reduction in AECB and the bacteria involved in the symptoms of AECB is observed.
[0101]
While preferred embodiments of the invention have been illustrated and described, it will be understood that various changes can be made therein without departing from the spirit and scope of the invention.
[Brief description of the drawings]
The above aspects and many of the attendant advantages of this invention will be more readily understood when the invention is better understood by reference to the above detailed description when taken in conjunction with the accompanying drawings. Is done.
FIG.
FIG. 1 shows the chemical structures of erythromycylamine and dirithromycin.
FIG. 2
FIG. 2 shows erythromycylamine hydrochloride in aqueous solution at 60 ° C., 100 mg / mL and 150 mg / mL and pH 5.0, pH 6.0 and pH 7.0 at 4 ° C. as described in Example 4. 5 is a graphical representation of the stability of the graph.
FIG. 3
FIG. 3 shows erythromycylamine hydrochloride in an aqueous solution of 60 mg / mL, 100 mg / mL and 150 mg / mL and pH 5.0, pH 6.0 and pH 7.0 at 25 ° C. as described in Example 4. 5 is a graphical representation of the stability of the graph.
FIG. 4
FIG. 4 shows erythromycylamine hydrochloride in an aqueous solution at 60 ° C., 100 mg / mL and 150 mg / mL and pH 5.0, pH 6.0 and pH 7.0 at 40 ° C. as described in Example 4. 5 is a graphical representation of the stability of the graph.
FIG. 5
FIG. 5 shows erythromycylamine hydrochloride in aqueous solution at 60 ° C., 60 mg / mL, 100 mg / mL and 150 mg / mL, and pH 5.0, pH 6.0 and pH 7.0 as described in Example 4. 5 is a graphical representation of the stability of the graph.
FIG. 6
FIG. 6 shows erythromycylamine sulfate in an aqueous solution of 60 mg / mL, 100 mg / mL and 150 mg / mL and pH 5.0, pH 6.0 and pH 7.0 at 60 ° C. as described in Example 4. 5 is a graphical representation of the stability of the graph.
FIG. 7
FIG. 7 shows erythromycylamine acetate at 60 ° C., 100 mg / mL and 150 mg / mL, and pH 5.0, pH 6.0 and pH 7.0 aqueous solutions at 60 ° C. as described in Example 4. 5 is a graphical representation of the stability of the graph.
FIG. 8
FIG. 8 shows a single intravenous dose of 25 mg / kg or a single inhalation dose of 30 mg / ml solution or 60 mg / ml solution over 30 minutes (0.7 mg / kg pulmonary dose or 5 shows the mean plasma concentration of erythromycilamine in rats (n = 3) after 1.77 mg / kg lung dose).
FIG. 9
FIG. 9 shows a single intravenous dose of 25 mg / kg or a single inhaled dose of a 30 mg / ml solution or a 60 mg / ml solution over 30 minutes (0.7 mg / kg pulmonary dose or 2 shows the average lung concentration of erythromycilamine in rats (n = 3) after 1.77 mg / kg lung dose).
FIG. 10
FIG. 10 shows 5 mg / mL (0.13 mg / kg), 25 mg / mL (0.27 mg / kg) and 50 mg / mL (50 mg / mL) for rats (n = 3) as described in Example 7. 1.3 mg / kg) after inhalation administration for 30 minutes a day containing an inhalation dose for 3 days. Figure 2 shows the efficacy of erythromycilamine in a pneumoniae lung infection model.
FIG. 11
FIG. 11 shows 1 mg / mL (0.03 mg / kg), 5 mg / mL (0.13 mg / kg), 25 mg / mL (25 mg / mL) for rats (n = 3) as described in Example 8. 0.27 mg / kg) and 50 mg / mL (1.3 mg / kg) after 30 minutes of inhalation administration as a single dose including inhalation doses. Figure 2 shows the efficacy of erythromycilamine in a pneumoniae lung infection model.
FIG.
FIG. 12 shows mean plasma and mean total lung concentrations of erythromycilamine after inhalation of a 60 mg / mL sulfate solution in dogs in a single dose for 30 minutes as described in Example 9. Show.
FIG. 13
FIG. 13 shows the average lung concentration of erythromycilamine in individual lung lobes after a single 30 minute inhalation dose of a 60 mg / mL sulfate solution in dogs as described in Example 9. .

Claims (38)

An aerosol formulation for the inhibition of susceptible bacteria in the endobronchial space of a subject suffering from an endobronchial infection, comprising:
The formulation uses a jet nebulizer, an ultrasonic nebulizer, a vibrating porous plate nebulizer or a dry powder inhaler capable of producing aerosol particles having a median aerodynamic diameter of between 1 μm and 5 μm in size. An aerosol formulation comprising from about 50 mg to about 750 mg of a macrolide antibiotic and a pharmaceutically acceptable carrier, which can then be administered in aerosol form. The aerosol formulation of claim 1, wherein the macrolide antibiotic is selected from the group consisting of erythromycylamine, dirithromycin, erythromycin A, clarithromycin, azithromycin, and roxithromycin. The aerosol formulation of claim 1, wherein the macrolide antibiotic is erythromycylamine. The aerosol formulation according to claim 1, wherein the formulation has a pH in the range of 5.0 to 7.0. The aerosol formulation according to claim 1, wherein the nebulizer is a jet nebulizer. The aerosol formulation according to claim 1, wherein the nebulizer is an ultrasonic nebulizer. The aerosol formulation according to claim 1, wherein the nebulizer is a vibrating porous plate nebulizer. The susceptible bacteria are Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Moraxella catarrhalis, Legionella pneumonia from the group of the sol. 9. The aerosol according to claim 8, wherein said pH is 6.0. The aerosol according to claim 9, wherein the nebulizer is a jet nebulizer. The aerosol according to claim 9, wherein the nebulizer is an ultrasonic nebulizer. The aerosol according to claim 9, wherein the nebulizer is a vibrating porous plate nebulizer. A method for treating an endobronchial infection of a susceptible bacterium by administering to a subject in need thereof an aerosol formulation for inhalation, comprising:
The formulation uses a jet nebulizer, an ultrasonic nebulizer, a vibrating porous plate nebulizer or a dry powder inhaler capable of producing aerosol particles having a median aerodynamic diameter of between 1 μm and 5 μm in size. And comprising about 50 mg to about 750 mg of the macrolide antibiotic and a pharmaceutically acceptable carrier, which can then be administered in aerosol form. 14. The method of claim 13, wherein the macrolide antibiotic is selected from the group consisting of erythromycylamine, dirithromycin, erythromycin A, clarithromycin, azithromycin, and roxithromycin. 14. The method of claim 13, wherein the macrolide antibiotic is erythromysylamine. 14. The method of claim 13, wherein the pH of the aerosol formulation ranges from 5.0 to 7.0. 14. The method according to claim 13, wherein the nebulizer used for administration of the aerosol formulation is a jet nebulizer. 14. The method of claim 13, wherein the nebulizer used for administration of the aerosol formulation is an ultrasonic nebulizer. 14. The method of claim 13, wherein the nebulizer used for administration of the aerosol formulation is a vibrating porous plate nebulizer. The susceptible bacteria are Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Moraxella catarrhalis, Legionella pneumonia from the group of the methods described in the group of the members of the group. 14. The method of claim 13, wherein a dose of less than about 2.0 ml of the nebulized liquid aerosol formulation comprising about 50 to about 150 mg / ml of the macrolide antibiotic is administered to the subject in less than about 10 minutes. Be done, way. 22. The method of claim 21, wherein the dose comprises less than about 1.5 ml of the nebulized aerosol formulation. 22. The method of claim 21, wherein the dose comprises less than about 1.0 ml of the nebulized aerosol formulation. 21. The method of claim 20, wherein the aerosol formulation comprises about 70 to about 130 mg / ml of the macrolide antibiotic. A unit dose device comprising a container comprising less than about 2.0 ml of a macrolide antibiotic formulation, comprising from about 50 to about 150 mg / ml of the macrolide antibiotic in a liquid physiologically acceptable carrier. , Unit dose device. 26. The unit dose device of claim 25, comprising about 1.5 ml of the macrolide antibiotic formulation. 26. The unit dose device of claim 25, comprising about 1.0 ml of the macrolide antibiotic formulation. 26. The unit dose device of claim 25, wherein the macrolide antibiotic formulation comprises about 70 to about 130 mg / ml of the macrolide antibiotic. 26. The unit dose device of claim 25, wherein the macrolide antibiotic formulation comprises about 90 to about 110 mg / ml of the macrolide antibiotic. 26. The unit dose formulation of claim 25, wherein the macrolide antibiotic is selected from the group consisting of erythromycylamine, dirithromycin, erythromycin A, clarithromycin, azithromycin, and roxithromycin. 26. The method of claim 25, wherein said macrolide antibiotic is erythromysylamine. 26. The unit dose device of claim 25, comprising less than about 2.0 ml of the macrolide antibiotic formulation, comprising about 20 to about 200 mg / ml erythromycylamine. A unit dose device comprising a container containing a macrolide antibiotic formulation, comprising from about 25 to about 250 mg of the macrolide antibiotic in a dry powdered physiologically acceptable carrier. 34. The unit dose device of claim 33, wherein the macrolide antibiotic formulation comprises about 50 to about 200 mg of the macrolide antibiotic. 34. The unit dose device of claim 33, wherein the macrolide antibiotic formulation comprises about 75 to about 150 mg of the macrolide antibiotic. 34. The unit dose device of claim 33, wherein the macrolide antibiotic is selected from the group consisting of erythromycylamine, dirithromycin, erythromycin A, clarithromycin, azithromycin, and roxithromycin. 34. The unit dose device of claim 33, wherein said macrolide antibiotic is erythromycylamine. 34. The unit dose device of claim 33, wherein the macrolide antibiotic formulation comprises about 50% to about 90% by weight of the macrolide antibiotic.

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