JP3840515B2 - Anti-asthma combinations containing surfactant phospholipids - Google Patents

Description

【００４４】
リン脂質組成物の界面活性度の測定
２ｃｍ×２ｃｍの白金灰色浸漬板をブンゼンバーナーの炎または同様の炎を用いてチェリーレッドになるまで加熱する。その板を５００ｍｇまで計量できる電子天秤から吊るす。
【００４５】
その装置を検定するために、小さいテフロン皿を、約２０℃（室温）の蒸留水で満たし、浸漬板の真下の実験用ジャッキの上に置く。そして、その皿を、浸漬板が水の表面を底端に沿って均一に丁度接するように持ち上げる。浸漬した場所で引き上げられているメニスカスにより電子天秤を、ペンレコーダーの表示を読むと約７３ｍNm^−１（２０℃における水の空気／水表面張力）となるように設定する。テフロン皿を下に降ろし、中を空にし、洗い、乾燥し、そして試薬級のメタノールで満たす。浸漬板は、上記のようにして洗浄される。そしてその皿を、浸漬板がメタノールの表面を底端に沿って均一に丁度接するように持ち上げる。浸漬した場所で引き上げられているメニスカスは、ペンレコーダーの表示を読むと約２２ｍNm^−１（２０℃におけるメタノールの空気／メタノール表面張力）である。テフロン皿を下に降ろし、浸漬板を上記の様に洗浄する。洗浄された板が単独の状態（即ち空気中で吊り下げられて）でゼロになければならない。
【００４６】
物質の界面活性度の量の測定値を得るために、テフロン皿は約３７℃まで暖められ、３７℃以下の水で満たされ、洗浄された板の真下の実験用ジャッキの上に置かれる。そして、その皿を浸漬板が水の表面を底端に沿って均一に丁度接するように持ち上げる。浸漬した場所で引き上げられているメニスカスの測定値は約７０ｍNm^−１（温水のおおよその空気／水表面張力）である。物質は小さな薬さじを用いて水の表面の上に広げられる。広げられた量は、過剰分（小さい自由浮遊粒子として）を観測できるように、水の表面上に完全な単一層が形成されることを確認できる十分な量であるべきである。表面張力は直ちに低下し、その低下はペンレコーダーによって記録される。平衡表面張力は約１分後に測定される。測定直後のテフロン皿中の水の温度は、３５℃以上であるであるべきである。
【００４７】
ここで用いられる「高界面活性度」の用語は、本発明に従い用いられるすべての組成物に関して、平衡表面張力が、上記の方法での測定により、水表面に組成物が広げられる前の表面張力より少なくとも１０％低いことを意味するものである。実際、薬剤（ａ）の説明において上述した様な、あるリン脂質組成物を使用することよる表面張力の減少は、５０％を超えるかもしれない。
【００４８】
他の物質を混合した状態で含まれる成分は、もし、混合物を用い、分離して他の物質を単独で用いて、表面張力の測定として上記の方法を実行した際において、平衡界面張力に至るまでにかかる時間が、混合物として物質単独と比較してより短くなる場合には、他の物質が拡散するのを高めることが理解されるべきである。
【００４９】
上記の方法は、３７℃での表面張力の測定を記述したものである。ここで、ほぼ通常の哺乳類の体温における拡散の高まりを参照したい場合には、通常の体温が約３７℃でない場合は、当該哺乳類のほぼ通常の体温においてその方法が実施されるべきである。
【００５０】
下記の実施例は、上皮に対する好ましいリン脂質の結合を説明したものである。
実施例
試薬
Ｌ−α−ホスファチジルコリン、１，２−ジ[１−^１４Ｃ]パルミトイル トルエン中：エタノール（１：１ Ｖ／Ｖ）、１１４ｍＣｉ／mmol、２ｍＬ中５０μＣｉ（ＣＦＡ６０４Ｂ３６、Ａｍｅｒｃｈａｍ）
Ｌ−α−ホスファチジルコリン、ジパルミトイル（Ｃ１６：０）（Ｐ−６２６７、シグマ）
ＤＬ−α−ホスファチジル−ＤＬ−グリセロール、ジパルミトイル（Ｃ１６：０）（Ｐ−５６５０、シグマ）
卵ホスファチジルグリセロール（Ｂａｔｃｈ２４７５６、Ｍａｃｆａｒｌｅｎ Ｓｍｉｔｈ、Ｌｔｄ）
塩化ナトリウム ０．９％ Ｂ．Ｐ．（バクスターヘルスケア）
塩化カルシウム （Ｃ−４９０１、シグマ）
トルエン（Ｔ−４４２８、シグマ）
エタノール ＡｎａｌａＲ（１０１０７．７Ｙ、ＢＤＨ）
ＮＣＳ−ＩＩ組織可溶化剤、０．５Ｎ溶液（ＮＮＣＳ−５０２、Ａｍｅｒｓｈａｍ）
ＯＣＳ 有機計算用シンチレーション剤（ＮＯＣＳ１０４、Ａｍｅｒｓｈａｍ）
培養された上皮の分散体の調製において、リン脂質組成物の貯蔵溶液は実施１の最初の日に調製された。これらの液は下記の通りである。
【００５１】
Ｌ−α−ＤＰＰＣ、２．４ｍｇ ｍＬ^−１ トルエン中 ：エタノール、１：１ＤＬ−α−ＤＰＰＧ、３．０ｍｇ ｍＬ^−１ トルエン中：エタノール、１：１卵 ＰＧ、３．０ｍｇ ｍＬ^−１ トルエン中：エタノール、１：１
【００５２】
上記の溶液全ては、ガラスバイアル中に４℃で保存され、そのねじ部は溶媒の蒸発を最小限にするためにテフロンテープで封じられている。そして、各ガラスバイアルは硬く蓋をした第２のガラスバイアルの内側に置かれた。これらの液は、試験における５つの実施のそれぞれに用いられ、０．９％生理食塩水中に塩化カルシウムが２００ｍｇＬ^−１の溶液は、実施２の初日に調製され、実施２から５のそれぞれにおいて用いられた。
【００５３】
装置
特製超音波洗浄機、型 Ｇｌ１２ ＳＰｌＧ（Ｌａｂｏｒａｔｏｒｙ Ｓｕｐｐｌｉｅｓ Ｃｏ．Ｉｎｃ、Ｈｉｃｋｓｖｉｌｌｅ、Ｎ．Ｙ．、ＵＳＡ）
ＶＦ２ ボルテックス（ＩＫＡ−Ｌａｂｏｒｔｅｃｈｎｉｋ）
撹拌水浴、型ＴＳＢ２−２０１−Ａ（Thermoline Scientific Equipment、Smithfield、オーストラリア）
Conthermシリーズ５、ファンオーブン（Contherm Scientific Ltd. Lower Hutt、ニュージーランド） TＲＩ−ＣＡＲＢ ２７００ＴＲ 液体シンチレーション分析器（Packard Instrument Co.、Meriden、CT、Ｕ．Ｓ．Ａ）
超音波洗浄機，型ＦＸＰＩ２（Unisonics Ｐｔｙ．Ｌｔｄ．シドニー、オーストラリア）
【００５４】
気管支の上皮
気管支上皮の提供源を与えるため、ブタの肺を死後２４時間以内に屠殺場から得た。その肺は死後４℃で保存されていたものである。第２の気管支は右及び／または左の肺から切り離された。気管支の表面の外側は全ての肺組織から削られ、その気管支を、前もって測定された気管支上皮の表面積（約０．５ｃｍ×０．５ｃｍ）を持つ切片に切断した。ただし、気管支と軟骨には手をつけずに残した。上皮の表面は粘液を取り除くために０．９％の生理食塩水ですすがれた。必要な場合には、気管支上皮の切片は使用のため必要となるまで０．９％生理食塩水で−２０℃で３から７日保存された。切片は各実施例の初日の使用前に解凍した。
【００５５】
気管支の上皮として、合計で５つの実施が行われた。各実施は、下記のように、３つのグループからなるものであった。
１．ＤＰＰＣのみ
２．ＤＰＰＣ＋ＤＰＰＧ
３．ＤＰＰＣ＋卵ＰＧ
【００５６】
４種の分散体は各実施における初日に調製された。全てのグループには、貯蔵溶液から^１４Ｃ−Ｌ−α−ＤＰＰＣの２０．５μＬ（３．３μｇ）と標識されていないＬ−α−ＤＰＰＣの５．５μＬ（１３．２μｇ）の両方を割り当てた。加えて、５．５μＬ（１６．５μｇ）のＤＬ−α−ＤＰＰＧがグループ２に割り当てられ、一方同量の卵ＰＧがグループ３に加えられた。グループ２と３において、ＰＧに対するＤＰＰＣ全量の比率は１：１であった。リン脂質成分に、グループ１、２及び３に対して、６．６ｍｌの０．９％生理食塩水を混合した。各処置グループにおいて２つの上皮切片がある時までに、上記全容量が使用された。切片の数が増加すると全成分量は上記と同じ比率を保持したままで増加率に応じて増加した。表２は培養混合物への付加物をまとめたものである。
【００５７】
【表２】

【００５８】
リン脂質成分を水系媒体に溶解するために、４つの培養分散体のそれぞれは、４５分間音波処理され、混合するために１分間撹拌した。
【００５９】
各分散体から、２ロットの２．８ｍＬを２つのガラスバイアルに移した。気管支上皮の２つの切片の４つのグループとなるように、上皮の１つの切片をこれらの各分散体中で培養した。気管支の上皮は、培養が行われた所与の日の全てにおいて、単一のブタから得られたものであった。培養は、撹拌水浴中で３７℃で２４時間行われた。
【００６０】
グループ１の分散体の一部分は、ガラスシンチレーションバイアルに移され、オーブン中３７℃で２４時間培養された。これらの一部分は、検量線のための標準として用いられた。取られた他のグループの分散体からの一部分とこれらからのβ−カウントとを適合させることは、同量のＤＰＰＣを含むことの確認のために、グループ１の分散体からのこれらと比較された。
【００６１】
各実施の２日目において、上皮の切片は、培養分散体から取り除かれ、０．９％生理食塩水で２０回すすがれ、ゆるく付着しているリン脂質を取り除くために水浴中で３７℃まで加温された。上皮の粘液表面を機械的に破らぬように注意が払われた。そして、気管支上皮の各切片は、付属のカートリッジから取り除かれた。上皮の切片は、さらに、シンチレーションバイアル中の上皮に１．５ｍLの容量で加えられる可溶化剤による組織の消化を助けるために、より小さい断片に切断された。同容量の可溶化剤を標準の部分にそれぞれ加え、ブランクに加えた。全てのバイアルは内容物を混合するためにゆっくりかき混ぜ、５５℃までファン撹拌型対流オーブンで一晩（１８〜２０℃）加温した。
【００６２】
各実施の三日目において、１０ｍLの有機計測シンチレーション剤は、各シンチレーションバイアルに加えられ、混合するために３０秒間撹拌した。
【００６３】
各サンプルのβ−カウントと標準は、液体シンチレーション分析器を用いて測定された。二回目の計測は、最初の計測から７時間以内に行われた。もし、２つの計測値が近い場合には、最初の測定のみが検量線を作成し、試料を定量するのに用いられた。
【００６４】
検量線より、上皮の各切片に吸収された^１４Ｃ−ＤＰＰＣの質量は、計算された。各切片に吸収された合計ＤＰＰＣの質量を算定するために、各分散体における^１４Ｃ−ＤＰＰＣの量はＤＰＰＣの合計量の５分の１であることから、^１４Ｃ−ＤＰＰＣの質量は５倍された。結果を、上皮の平方センチあたりの吸収されたDPPCの合計量として表２に示した。
【００６５】
表３において、結果は、ＤＰＰＣの存在下において気管支の上皮に対するＤＰＰＣの結合の増加が観測されたが、結合の程度はＤＰＰＧではなく卵ＰＧを用いた範囲に比べて更に改善されたことを示している。
【００６６】
本発明は喘息に対する人間の患者の治療に関し特に述べてきたが、本発明を例えば鼻炎といった他の肺疾病の治療に適用することもできる。
【００６７】
本発明の組合せ製品は、人間以外の哺乳類の肺の状態を治療するのに使用することができる。例としては、馬の反応性気道疾病が挙げられる。
【００６８】
【表３】

【図面の簡単な説明】
【図１】デリバリー装置の横立体図。
【図２】図１と同様の図で、その内部を示したもの。
【図３】本発明によるデリバリー装置の別の態様の概念図。[0001]
The present invention relates to a pharmaceutical product for use in the treatment of asthma and a delivery device comprising the product.
[0002]
Asthma is estimated to be between 4 and 10 percent of the population and causes bothering and alertness for both patients and those around them. The onset of asthma appears to be often promoted by a number of factors, such as exercise and inhaled air pollutants. Other factors, such as pollen and airborne dust, make the respiratory tract sensitive and make it easier for asthmatics to get sick. This confirms that effective treatment can include drugs that reduce the sensitivity of asthma patients to allergens or drugs that neutralize allergic reactions.
[0003]
Non-asthmatic lungs and airways appear to contain natural barriers that prevent pollutants and other potential irritants from reacting with receptors that cause acute attacks. The binding of surface active phospholipids (SAPL) to the surface tissue of the lungs reduces the number of receptors exposed to harmful stimuli and reduces the reflex of bronchoconstriction, which is normal in the lungs of asthmatic patients. Studies suggest that the situation in the lungs can be mimicked.
[0004]
Bangham et al. , Colloids & Surefaces, 10 (1984), 337-341, SALP is clinically used as a treatment for respiratory urgency syndrome (RDS) in neonates. In this role, SAPL is considered to function by reducing the high interfacial tension acting on the air-water interface in the alveoli, thereby reducing the pressure required to expand the lungs. Accordingly, commercially available formulations of SAPL are designed to spread rapidly over the air-water interface, thereby reducing the extremely high interfacial tension of water.
[0005]
Kurashima et al Jap. J. et al. As seen in Allergor 1991; 40,160, clinical studies have been conducted to determine the effects of commercial SAPL, which is limited, but marketed as a treatment for neonatal RDS in asthma issues. It is coming. This paper reported some improvements in bronchoconstriction in asthmatic adults using SAPL obtained by extracting from bovine lungs. In another study on children, as seen in Oetomo et al American Journal of Respiratory and Critical Care Medicine 153; 1996, 1148, pulmonary function or histamine response was also obtained using SAPL extracted from bovine lung. No significant changes were observed in
[0006]
EP 0 528 034A describes the use of pulmonary surfactant as an anti-asthma component in the form of a liquid or suspension for injection or spraying into the patient's respiratory tract.
[0007]
The present invention is a therapeutic combination product for use in the prevention and / or treatment of asthma comprising:
(A) A composition comprising a surface-active phospholipid (SAPL) in a finely divided shape, and SAPL contains a component that promotes the spread of the drug on the surface at almost normal mammalian body temperature. Yes, and
(B) Anti-asthma drugs
Including
Component (a) and component (b) are provided together or separately in dosage form.
The finely divided powder of component (a) preferably contains at least a first and a second component, but is considered to have two important effects.
[0008]
First, the drug (a) has a surfactant property that allows it to spread rapidly on the surface of the lungs and airways. It is an important feature in the present invention that the drug (a) is in the form of a powder, that is, in the form of a solid. This “dry” surfactant has a high surface activity. In contact of the drug (a) which is the first component with the mucus in the lung, the presence of the second component lowers the melting point of the first component, and the first component rapidly forms on the liquid-air interface as a thin film at body temperature. It is thought to bring about a result that promotes the spread. For example, the normal melting point of dipalmitoylphosphatidylcholine, which is preferred as the first component, is about 40 ° C., that is, above normal body temperature. However, when used in combination with a suitable second component such as phosphatidylglycerol, the melting point of dipalmitoyl phosphatidylcholine can be lowered substantially below normal body temperature.
[0009]
Secondly, once the surface active agent covers the lungs and airway surfaces in situ, the components of the composition migrate beyond the mucus layer, leaving a thin hydrophobic inner layer or coating on the tissue surface. It is thought to adsorb on the top. Thus, on top of the above-described features that reduce surface tension, the agents of the present invention are believed to provide a protective effect, thanks to the adsorption layer. In binding to the epithelium, the phospholipids will mask the stimulant receptors that manifest the bronchoconstriction reflex, ie narrow the bronchi.
[0010]
The drug (a) is in the form of a finely divided solid. As a result of the high surfactant activity of the drug (a) in its form, a large reduction in surface tension in contact with the lung mucus layer results, which increases the effectiveness of the ingredients and the lungs of the asthma drug being dosed It is thought to improve access to the surface. Therefore, the use of drug (a) in combination with anti-asthma drugs is thought to extend the effectiveness of anti-asthma drugs.
[0011]
Furthermore, as noted above, binding of the phospholipid component to the lung surface is believed to reduce bronchoconstriction as a result of a decrease in receptor-mediated activity that has the property of shielding the stimulant receptor. Its reduction in bronchoconstriction acts cumulatively with the anti-bronchial contractile activity of anti-asthma drugs. Thus, as a result of the synergistic effect of drug (a) in improving the effectiveness of anti-asthma drugs, as well as the drug's own additional anti-bronchial contractile activity, depending on the situation, It may also be possible to reduce the dose of anti-asthma medication.
[0012]
As used herein, “finely divided” refers to a substance having a particle size distribution such that at least a large percentage of the weight of the particles enters the patient's respiratory tract and is probably small enough to enter deeply into the lungs when inhaled. In fact, the first component and the second component preferably have a particle size distribution of 90% by weight or more, and a particle size of 10 μm or less, particularly 5 μm or less. preferable. Advantageously, the median particle size of the combination of the first component and the second component is more preferably the median particle size of each of the first component and the second component is 10 μm or less, more Preferably it is 5 micrometers or less. The median particle size may be less than 3 microns, for example about 1.2 microns. In some circumstances it is preferred that the particles have a median particle size of at least 0.5 μm. The particle diameter can be calculated by laser diffraction, and can also be calculated by other methods that can determine the aerodynamic average value of the particles. Here, the median average particle diameter means a weight median aerodynamic particle diameter (MMAD). MMAD can be obtained in any suitable manner, for example, European Pharmacopoeia (Appendix 1999) 2.9.18. It can be determined by a multi-stage liquid impedance according to the method described in (Aerodynamic Evaluation of Fine Particles). Alternatively, the particle size distribution can be characterized by a volume average particle size (VMD). Advantageously, the VMD is 10 μm or less, for example 5 μm or less, preferably less than 3 μm. It is believed that this type of finely divided dry particles, referred to as fumed particles, can be adsorbed to the surface of lung tissue and in use will reach the epithelium.
[0013]
The finely divided solid mixture of the first component and the second component of the drug (a) is preferably suitable for reducing the diameter by reducing the size of the larger particles before mixing. Any method can be used. Preferably, the first component of the drug (a) contains one or more compounds selected from the group consisting of diacylphosphatidylcholines. Examples of suitable diacylphosphatidylcholines (DAPCs) are dioleyl phosphatidylcholine (DOPC), distearyl phosphatidylcholine (DSPC) and dipalmitoylphosphatidylcholine (DOPC). Each of these compounds is known to form a thin film on or cover the lung surface. Most preferably, the first component is DPPC. The second component is preferably composed of one or more compounds selected from the group consisting of phosphatidylglycerol (PG), phosphatidylserine (PS), phosphatidylinositol (PI) and chlorestyl palmitate (CP).
[0014]
Phosphatidylglycerol (PG) is thought to be able to bind to lung tissue and enhance binding with the first component and is therefore a preferred second component. PG is also a preferred second component by having the ability to form a dispersion of very finely divided dry particles in air with the first component.
[0015]
Advantageously, the drug comprises diacyl phosphatidylcholine and phosphatidylglycerol. Advantageously, the phosphatidylglycerol is a diacylphosphatidylglycerol. The acyl group of phosphatidylglycerol, whether the same or different, is advantageously a fatty acid acyl group having 14 to 22 carbon atoms, respectively. Indeed, the phosphatidylglycerol component may be a mixture of phosphatidylglycerols containing different acyl groups. Phosphatidylglycerol can be conveniently obtained by synthesis from purified lecithin, and the constituents of the acyl substituent depend on the source of lecithin used as the raw material. At least a portion of the fatty acid acyl group of the phosphatidylglycerol is preferably an unsaturated fatty acid residue, such as a mono- or diunsaturated C18 or C20 fatty acid residue. Preferred acyl substituents in the phosphatidylglycerol component are palmitoeoyl, oleoyl, linoleoyl, linolenoyl, arachidonoyl groups. Preferably, the medicament comprises dipalnitoyl phosphatidylcholine and phosphatidylglycerol so as to have a phosphatidyl moiety of phosphatidylglycerol which can be advantageously obtained from the phosphatidyl part of egg lecithin.
[0016]
The first component and the second component of drug (a) may be present in a weight ratio of 1: 9: to 9: 1. Advantageously, the proportion by weight of the first component is greater than that of the second component. Preferably, the first component and the second component are present in a weight ratio of 6: 4 to 8: 2. At a weight ratio of about 7: 3, the mixture diffuses rapidly at a temperature of 35 ° C. or higher.
DPPC is available from Can. J. et al. Of Biochem. Physiol 1959, 37, page 953, Baer & Bachrea, can be obtained synthetically by acylation of glyceryl phosphoryl chloride and is commercially available from Sigma (London). PG is described in Comfurions et al, Biochem. Bophys Acta 1977, 488 pages 36-42 and Dawson, Biochem J. Biol. 1967, 102, from page 205 to page 210 and can be obtained from egg phosphatidylcholine. When coprecipitated with DPPC from a common solvent such as chloroform, PG forms fine particles that rapidly diffuse with the DPPC over the airway and lung surfaces. Most preferably, the composition of the present invention comprises DPPC and phosphatidylglycerol obtained from egg phosphatidylglycoline and having a mixture of C16, C18 (saturated and unsaturated) and C20 (unsaturated) acyl groups. preferable. One form of this composition is Britannia Pharmaceuticals Ltd. , 41-51 brightton Road, Redhill, Surrey, under the trademark “ALEC”. However, for use in the apparatus of the present invention, it is preferred that the particle size of the mixture be smaller than the currently marketed form of “ALEC”. In order to obtain a mixture having a particle size suitable for use in the apparatus of the present invention, the phospholipid component is dissolved in a suitable solvent such as ethanol, the solution is filtered and vacuum dried, and further desired. The particle size of the solid product is reduced in order to obtain particles with a diameter of. Care must be taken during particle size reduction to prevent the mixture from moisture, oxygen, direct heating, static electricity, and microbial contamination. Here, the “anti-asthma drug” is used to include any drug having biological activity against asthma. As used herein, an “anti-asthma drug” would be understood to be free of the drug component (a) of the composition. The anti-asthma drug includes one selected from the group consisting of β2-agonists, steroids, chromones, antimuscarinic drugs, and leucotrin receptor antagonists, and is one or more of respiratory drugs The above may be included. A combination product comprises one or more anti-asthma drugs in an amount up to 10 parts by weight per 100 parts by weight of the combination of the first and second components of the drug (a), especially One part by weight can be included as an upper limit with respect to 100 parts by weight. Desirably, the respiratory drug or plurality of respiratory drugs are provided in an amount such that each dose delivered by the device comprises an effective amount of the respiratory drug or the plurality of respiratory drugs.
[0017]
The combination product may include a β2 agonist, which may be a terbutaline, eg, a terbutaline salt such as terbutaline sulfate or a combination thereof, or salbutamol, salbutamol salt, or a combination thereof. Salbutamol and its salts are widely used in the treatment of respiratory diseases. The active particles may be salbutamol sulfate particles. Long-acting β2 adrenergic receptor agonists may be present, for example, with formoterol, salmeterol, or salts thereof.
[0018]
The combination product may include an antimuscarinic agent such as ipatropium bromide.
The combination product may include a steroid, which may be, for example, beclomethasone dipropionate, budesonide, triamcinolone acetonide, or fluticasone. The drug may include other preventive drugs, and the other preventive drugs include chromones such as sodium cromoglycate and nedocromil. This drug may include a leucotrin receptor agonist.
[0019]
Advantageously, at least component (a) is arranged for delivery to a patient in the form of a single person inhalable dose, the single individual or each individual's dose being said first component of component (a) And the second component in a combined amount of at least 10 mg. Although phospholipids have already been disclosed as adjuncts in certain forms of delivery devices, the amount of phospholipids to be administered in doses with these already disclosed devices is much higher than that predicted by following the present invention. There were few. Indeed, according to the present invention, each individual dose is preferably at least 25 mg, more preferably at least 40 mg of the first and second components. The first component and the second component are substantially non-toxic, and the upper dosage limit of component (a) is therefore generally convenient, taking into account, for example, patient comfort and / or device design parameters. It is selected in consideration of sex. In general, however, the device is capable of delivering doses up to 1000 mg, more advantageously up to 500 mg, preferably up to 200 mg, more preferably up to 100 mg. Preferably at least component (a) is arranged for multiple sequential delivery of the doses that can be taken.
[0020]
The product of the present invention has the further advantage that the first and second components of drug (a) may be derived from synthesis. Exposure of asthma patients to animal-derived proteins has been found to be unfavorable because they can be hypersensitive to patients, and therefore the use of synthetic substances is not recommended for animal-derived surfactants that may contain animal protein. Is very advantageous.
[0021]
In the present invention, it is highly desirable that the drug (or any active ingredient) does not disrupt the lung environment, since it is desirable to achieve absorption over a relatively long period of the drug (a) on the lung surface. One factor that reduces the lifetime of the inner layer or coating is the presence of an enzyme such as phospholipase A that can digest DPPC and / or PG. Such an enzyme attacks only the levorotatory (L) body, and this levorotatory (L) body constitutes a naturally obtained form. Therefore, the drug includes a dextrorotatory (D) isomer or at least a racemic isomer, and the dextrorotatory (D) isomer is preferably obtained by a synthetic route.
[0022]
A suitable dispersing device may use a propellant such as a halocarbon to form a gas stream, accelerate the particles through a tapered discharge nozzle, and discharge nozzle baffles to remove particles that are too large, A venturi may be provided. Suitable halocarbons include low-boiling hydrofluorocarbons, hydrofluorochlorocarbons, fluorochlorocarbons, for example, these are marketed under the trademark “Freon”. The drug may be enclosed with a propellant in a pressurized aerosol container in the inhaler. For example, as seen in US Pat. No. 5,577,497, other inhalers include a propellant that mixes powder into an air stream and delivers the powdered air to the patient's respiratory tract. It is.
[0023]
Preferred methods and devices for dispensing drug (a) include those that disperse the powdered drug in a propellant gas stream. For example, a pressurized liquefied gas filter box is preferably connected to a vial containing a drug. By releasing a regulated amount of gas from the filter box into the vial, the drug increment is expelled from the vial as a cloud of powder and is inhaled by the user. If the characteristics of the anti-asthma drug administered together are compatible, the drug can be introduced into a gas stream and mixed with the drug (a) for administration. In use, one or two inhalable doses of drug (a) each contain 50 mg, but may be taken up to 3 times a day.
[0024]
Depending on the patient's requirements, anti-asthma drugs may be separated and administered sequentially with drug (a). Therefore, the timing of the medication does not depend on the timing of the drug (a).
[0025]
The present invention provides a delivery device for administering a drug for prevention or treatment of asthma to a patient by inhalation, and this delivery device is a finely divided form of a surface active phospholipid (SAPL). And) the SAPL comprises a component that enhances drug diffusion and the delivery device is capable of delivering to at least one individual in an amount of at least 10 mg.
[0026]
The present invention also provides a delivery device for administering a drug for prevention or treatment of asthma to a patient by inhalation, the delivery device containing the drug, and the drug is finely divided One or more ingredients selected from the group consisting of a first ingredient comprising one or more phosphatidylcholines and phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and chlorestil palmitate, in powder form A second component comprising: a delivery device, wherein the delivery device is arranged for delivery of a dose that can be aspirated by at least one individual, wherein the single dose or each individual dose is the first phospholipid; Including the component and the second component in a combined amount of at least 10 mg That.
[0027]
Furthermore, the present invention provides the use of (b) a surfactant phospholipid (SAPL) component that is in a finely divided form combined with an anti-asthma drug in the manufacture of a medicament for regulating asthma It is.
[0028]
Now, one form of distributor according to the present invention will be described in detail by way of illustration with reference to the accompanying drawings. The figure is:
FIG. 1 is a horizontal three-dimensional view of the delivery device,
FIG. 2 is a similar view, but showing its interior, and
FIG. 3 is a conceptual diagram of another embodiment of the delivery device according to the present invention.
[0029]
In the drawing, the casing 1 is formed by two plastic molds 2 and 3 that mesh with each other to form a container for the pressure filtration device box 4 and the vial 5. The pressure filtration unit box 4 contains a low boiling point liquid, preferably a hydrofluorocarbon such as HFA-134a or HFC-227, under sufficient pressure to maintain the propellant as a liquid at normal room temperature. The vial 5 contains a powder drug (a) such as “ALEC”. The filtration device 4 is provided with a release valve 6 that is received in the recess 7 so that the finger pressure applied to the reverse end 8 releases the propellant to the tube 9. The tube 9 is typically made of hard plastic, such as PVC or polypropylene, and is a tube having an outer diameter of about 2-3 mm and an inner diameter of about 0.5-2 mm. The tube 9 is connected to the valve 6 by a fitting 10 and from there to a tube or needle 11 extending into the vial 5. The vial 5 may be closed by a rubber sealing material through which the tube or needle 11 penetrates and self-sealing around the tube or needle. A second needle or tube 12 extends through the rubber seal at the neck of the vial and extends partway through the vial and is connected to the fitting 13. The mounting part 13 is discharged into a mouthpiece 14 that is shaped to be easy for the user to place in the mouth. When the patient needs the drug, the patient places the mouthpiece 14 in the mouth, breathes, and simultaneously depressurizes the filter device box 4. This causes a cloud of medication to be dispensed into the patient's respiratory tract. The attachment parts 10 and 13 may be valve bodies. The valve body 10 is installed to allow a metered amount of propellant to enter the vial. Similarly, the valve body 13 is installed for releasing when the pressure in the vial reaches a predetermined level. This inhaler is preferably used with one hand in a manner similar to conventional nebulizers.
[0030]
The anti-asthma drug may be administered separately from the separation apparatus either immediately before or after the administration of the drug (a), or may be administered separately according to the patient's request. The anti-asthma drug may be dispensed from any suitable type of suction device, such as, for example, a dry powder inhaler or a pressurized medication inhaler. Such devices with anti-asthma drugs are well known and widely commercially available and do not require further explanation.
[0031]
Alternatively, in addition to the powdered phospholipid composition, vial 5 may be integrated with other known lung or respiratory drugs, such as salbutamol, beclomethasone, corticosteroids or other asthma drugs. However, it is preferred to enclose conventional asthma drugs in a propellant filtration device box or in a capsule sandwiched between a propellant container and a vial containing a phospholipid composition. In this method, the lungs and airways receive a cloud of phospholipid composition and conventional aerosols, either sequentially or simultaneously. This combination therapy rapidly removes pain and continues protection as a thin layer of phospholipid spreads over lung tissue. Instead of encapsulating the phospholipid composition in a multipurpose vial, it may be included in a single-use capsule between the outlet from the propellant filtration device and the mouthpiece.
Another form of delivery device is illustrated in FIG.
[0032]
Conceptually, the device 101 shown in FIG. 3 provides a container 102 having a volume of several liters filled with an aerosolizable solid SAPL composition and optionally also containing an anti-asthma drug. And is aspirated by the patient via a breathing tube 120 connected to a tube 104 leading from the container. The container 102 is first evacuated using a vacuum pump 115. A certain amount of solid, which is a powder SAPL composition, is contained in a mesh holder 105 in the tube 106 and air is used to form a cloud that has been made aerosolizable in the container 102 by air. Introduced through. When the container 102 reaches approximately atmospheric pressure, the respiratory tube 102 reaches approximately atmospheric pressure, and the respiratory tube 120 is opened to allow the patient to aspirate its SAPL composition.
[0033]
The apparatus 101 includes a stainless steel container 102 having a capacity of about 4 liters and having an opening 103 connected to a vertically extending pipe 104 at the upper end. The tube 104 is connected to a sideways tube 109 and a breathing tube 120 extending through the screen 121 so that the device is not visible to the patient. The breathing tube 120 may be mated with a plug at the far end, and the plug is removed prior to use. The mesh holder 105 is installed on the top of the container 102 as a part connected between the air line 106 and the container. The mesh holder can also be removed to introduce a quantity of powdered drug into the delivery device. One end of the air line 106 is connected to the container 102 via a part 103 via a mesh holder. The other end of the air line 106 is connected via a regulating device 107 to a regulated propellant 108 of compressed propellant such as air. If necessary, the source of the compressed propellant can also include biologically active ingredients for the treatment of asthma. Tube 104 extends upward from the container to intersect horizontally extending tube 109 and extends from one end of tube 110 extending to the atmosphere. A valve body 111 that can be opened and closed by a handle 112 is provided in a horizontally extending pipe 109 and closes the pipe 110 from the container 102 except when the valve body 111 is opened.
[0034]
At the other end of the horizontal tube 109, a pressure gauge 113 is prepared. At its end, the horizontal tube 109 is connected to an air line 114 that extends to the vacuum pump 115 via the regulator 107, which can be adjusted independently of the regulator 107. The valve body 116 can be operated by a handle 117 and is prepared for opening and closing a pipe between the container 102 and the air line 114.
The safety pressure relief valve body 118 is integrated with the device and is arranged to open when it is 0.034 bar above atmospheric pressure.
[0035]
A SAPL composition that is micronized in use (optionally with an anti-asthma drug) may be introduced into the mesh holder device 105, which is inserted into the mouth 103 following the container 102. Has been. Upon insertion of the mesh holder device, the container is sealed and both valve bodies 111 and 116 are closed. The pressure inside the container 102 is then reduced by opening the valve body 116 and exhausting the air in the container 102 through the air line 114.
[0036]
The adjustment unit 107 may include a needle valve (which can be adjusted) for adjusting the rate of exhausting air from the container 102. When the pressure in the container drops too rapidly, the powder medicine in the mesh holder device may be drawn into the container at an early stage.
Thereafter, the valve body 116 is closed. While the container 102 is sealed with reduced internal pressure, the regulated compressed air supply 108 is temporarily moved to inject air through the mesh holder device 105 into the container 102. As a result, the powder in the mesh holder device 105 is aerosolized and enters the container 102. The pressure is measured using a pressure gauge 113, and at this stage it must be at or slightly lower than atmospheric pressure.
[0037]
The plug is then removed from the mouthpiece of the respiratory tract, and the patient can aspirate the contents of the container by placing a mouth on the tip of the mouthpiece of the respiratory tract and inhaling.
[0038]
After the suction phase, the valve 11 may be closed and this circulation is started again.
[0039]
If necessary, the amount of well aerosolized powder should be weighed before use (mesh is pre-measured) and after use of the device. And the remaining powder can be measured by weighing.
[0040]
As indicated above, the anti-asthma drug may be present in the source of compressed propellant or may be placed in a mesh holder device with SALP.
[0041]
Where it is preferred or necessary for anti-asthmatic properties to be dosed by combination therapy with surface active phospholipids, anti-asthma medications are anti-aseptic, eg, dry powder inhalers or pressurized medication inhalers. It may be administered separately from other devices such as those well known for use in asthma medications.
[0042]
Measurement of fine particle classification of phospholipid composition
As already mentioned, the finely divided ALEC used in the product of the present invention is obtained by dissolving its components, filtering and vacuum drying, and the solids so obtained. It can also be obtained by reducing the diameter of the product. Delivery of reduced-size substances is described in European Pharmacopoeia (Appendix 1999) 2.9.18. According to the method described in (Aerodynamic Evaluation of Fine Particles), it is measured by a multistage impedance.
The vial with the substance is loaded into a five stage MLSI and the delivery of the substance is tested under many operating conditions. Each volume of 41 passed air is considered equal to the inhalation volume of one patient. In Table 1, the results show that relatively large respirable particles have been generated. The aspirable (or particulate) classifier shows particles that have reached MSLI 3, 4, and 5 stages, and the particle size of the particles is less than 5.3 μm. Such particles are considered to be those that penetrate deep into the typical patient's lungs.
[0043]
[Table 1]

[0044]
Measuring the surface activity of phospholipid compositions
A 2 cm x 2 cm platinum gray dip plate is heated using a Bunsen burner flame or similar flame until cherry red. The plate is hung from an electronic balance capable of weighing up to 500 mg.
[0045]
To calibrate the apparatus, a small Teflon dish is filled with distilled water at about 20 ° C. (room temperature) and placed on a laboratory jack just below the dip plate. The dish is then lifted so that the dip plate just touches the surface of the water evenly along the bottom edge. When the electronic balance is read by the meniscus pulled up at the place where it is immersed, the pen recorder reads about 73 mNm. ^-1 It is set to be (water air / water surface tension at 20 ° C.). The Teflon dish is lowered, emptied, washed, dried, and filled with reagent grade methanol. The immersion plate is cleaned as described above. The dish is then lifted so that the dip plate just touches the methanol surface evenly along the bottom edge. The meniscus pulled up at the place of immersion is about 22 mNm when reading the indication on the pen recorder. ^-1 (Methanol air / methanol surface tension at 20 ° C.). Lower the Teflon dish and wash the dip plate as above. The cleaned plate must be zero in a single state (ie suspended in air).
[0046]
To obtain a measure of the amount of surface activity of the material, the Teflon dish is warmed to about 37 ° C., filled with water below 37 ° C. and placed on a laboratory jack just below the cleaned plate. Then, the dish is lifted so that the immersion plate touches the surface of the water evenly along the bottom edge. The measured value of the meniscus pulled up in the immersed area is about 70 mNm. ^-1 (Approximate air / water surface tension of warm water). The substance is spread over the surface of the water using a small spoon. The spread amount should be sufficient to ensure that a complete monolayer is formed on the surface of the water so that an excess (as small free suspended particles) can be observed. The surface tension drops immediately and the drop is recorded by a pen recorder. The equilibrium surface tension is measured after about 1 minute. The temperature of the water in the Teflon dish immediately after the measurement should be above 35 ° C.
[0047]
As used herein, the term “high surface activity” refers to the equilibrium surface tension for all compositions used in accordance with the present invention, the surface tension before the composition is spread on the water surface as measured by the method described above. Means at least 10% lower. Indeed, the reduction in surface tension by using certain phospholipid compositions, as described above in the description of drug (a), may exceed 50%.
[0048]
Ingredients contained in a mixture of other substances will reach an equilibrium interfacial tension when the above method is performed as a measurement of surface tension using the mixture, separating and using the other substance alone. It should be understood that if the time it takes to become shorter as a mixture compared to the substance alone, it increases the diffusion of other substances.
[0049]
The above method describes the measurement of surface tension at 37 ° C. Here, if it is desired to refer to the increase in diffusion at approximately normal mammalian body temperature, if the normal body temperature is not about 37 ° C., the method should be carried out at approximately normal body temperature of the mammal.
[0050]
The following examples illustrate preferred phospholipid binding to the epithelium.
Example
reagent
L-α-phosphatidylcholine, 1,2-di [1- ¹⁴ C] Palmitoyl in toluene: ethanol (1: 1 V / V), 114 mCi / mmol, 50 μCi in 2 mL (CFA604B36, Amercham)
L-α-phosphatidylcholine, dipalmitoyl (C16: 0) (P-6267, Sigma)
DL-α-phosphatidyl-DL-glycerol, dipalmitoyl (C16: 0) (P-5650, Sigma)
Egg phosphatidylglycerol (Batch 24756, Macfarlen Smith, Ltd)
Sodium chloride 0.9% P. (Baxter Healthcare)
Calcium chloride (C-4901, Sigma)
Toluene (T-4428, Sigma)
Ethanol AnalaR (10107.7Y, BDH)
NCS-II tissue solubilizer, 0.5N solution (NNCS-502, Amersham)
OCS scintillation agent for organic calculation (NOCS104, Amersham)
In preparing the cultured epithelial dispersion, a stock solution of the phospholipid composition was prepared on the first day of Example 1. These liquids are as follows.
[0051]
L-α-DPPC, 2.4 mg mL ^-1 In toluene: ethanol, 1: 1 DL-α-DPPG, 3.0 mg mL ^-1 In toluene: ethanol, 1: 1 egg PG, 3.0 mg mL ^-1 In toluene: ethanol, 1: 1
[0052]
All of the above solutions are stored at 4 ° C. in glass vials and their threads are sealed with Teflon tape to minimize solvent evaporation. Each glass vial was then placed inside a second hard-capped glass vial. These solutions were used for each of the five runs in the study, with 200 mg calcium chloride in 0.9% saline. ^-1 Solutions were prepared on the first day of Example 2 and used in each of Examples 2-5.
[0053]
apparatus
Special ultrasonic cleaner, type Gl12 SPlG (Laboratory Supplements Co. Inc, Hicksville, NY, USA)
VF2 Vortex (IKA-Labortechnik)
Stir bath, type TSB2-201-A (Thermoline Scientific Equipment, Smithfield, Australia)
Contherm Series 5, Fan Oven (Contherm Scientific Ltd. Lower Hutt, New Zealand) TRI-CARB 2700TR Liquid Scintillation Analyzer (Packard Instrument Co., Meriden, CT, USA)
Ultrasonic cleaner, type FXPI2 (Unisonics Pty. Ltd. Sydney, Australia)
[0054]
Bronchial epithelium
To provide a source of bronchial epithelium, pig lungs were obtained from the slaughterhouse within 24 hours after death. The lungs were stored at 4 ° C after death. The second bronchus was dissected from the right and / or left lung. The outside of the bronchial surface was shaved from all lung tissue and the bronchi were cut into sections with the previously measured bronchial epithelial surface area (approximately 0.5 cm x 0.5 cm). However, the bronchi and cartilage were left untouched. The surface of the epithelium was rinsed with 0.9% saline to remove mucus. If necessary, bronchial epithelial sections were stored in 0.9% saline at −20 ° C. for 3-7 days until needed for use. Sections were thawed before use on the first day of each example.
[0055]
A total of 5 runs were performed as bronchial epithelium. Each implementation consisted of three groups as described below.
1. DPPC only
2. DPPC + DPPG
3. DPPC + egg PG
[0056]
Four dispersions were prepared on the first day of each run. All groups include stock solutions ¹⁴ Both 20.5 μL (3.3 μg) of C-L-α-DPPC and 5.5 μL (13.2 μg) of unlabeled L-α-DPPC were assigned. In addition, 5.5 μL (16.5 μg) of DL-α-DPPG was assigned to group 2, while the same amount of egg PG was added to group 3. In groups 2 and 3, the ratio of total DPPC to PG was 1: 1. The phospholipid component was mixed with 6.6 ml of 0.9% saline for groups 1, 2 and 3. By the time there were two epithelial sections in each treatment group, the full volume was used. As the number of sections increased, the total component amount increased according to the increase rate while maintaining the same ratio as above. Table 2 summarizes the additions to the culture mixture.
[0057]
[Table 2]

[0058]
In order to dissolve the phospholipid component in the aqueous medium, each of the four culture dispersions was sonicated for 45 minutes and stirred for 1 minute to mix.
[0059]
From each dispersion, two lots of 2.8 mL were transferred to two glass vials. One section of the epithelium was cultured in each of these dispersions to be four groups of two sections of bronchial epithelium. The bronchial epithelium was obtained from a single pig on all days of culture. Incubation was performed in a stirred water bath at 37 ° C. for 24 hours.
[0060]
A portion of the Group 1 dispersion was transferred to a glass scintillation vial and incubated in an oven at 37 ° C. for 24 hours. Some of these were used as standards for the calibration curve. Matching fractions from other groups of dispersions taken with β-counts from these were compared to those from dispersions of group 1 to confirm that they contained the same amount of DPPC. It was.
[0061]
On the second day of each run, epithelial sections are removed from the culture dispersion and rinsed 20 times with 0.9% saline and heated to 37 ° C. in a water bath to remove loosely attached phospholipids. Warmed. Care was taken not to mechanically break the mucus surface of the epithelium. Each section of bronchial epithelium was then removed from the attached cartridge. Epithelial sections were further cut into smaller pieces to aid in digestion of the tissue by solubilizers added in a volume of 1.5 mL to the epithelium in scintillation vials. The same volume of solubilizer was added to each standard part and added to the blank. All vials were stirred gently to mix the contents and warmed to 55 ° C. overnight (18-20 ° C.) in a fan stirred convection oven.
[0062]
On the third day of each run, 10 mL of organic counting scintillation agent was added to each scintillation vial and stirred for 30 seconds to mix.
[0063]
The β-count and standard of each sample was measured using a liquid scintillation analyzer. The second measurement was performed within 7 hours from the first measurement. If the two measurements were close, only the first measurement was used to create a calibration curve and quantify the sample.
[0064]
Absorbed by each section of epithelium from calibration curve ¹⁴ The mass of C-DPPC was calculated. To calculate the mass of total DPPC absorbed in each section, in each dispersion ¹⁴ Since the amount of C-DPPC is one fifth of the total amount of DPPC, ¹⁴ The mass of C-DPPC was multiplied by 5. The results are shown in Table 2 as the total amount of DPPC absorbed per square centimeter of epithelium.
[0065]
In Table 3, the results show that increased binding of DPPC to bronchial epithelium was observed in the presence of DPPC, but the degree of binding was further improved compared to the range using egg PG rather than DPPG. ing.
[0066]
Although the present invention has been specifically described with respect to treating human patients for asthma, the present invention can also be applied to the treatment of other pulmonary diseases such as rhinitis.
[0067]
The combination product of the present invention can be used to treat lung conditions in non-human mammals. Examples include reactive airway disease in horses.
[0068]
[Table 3]

[Brief description of the drawings]
FIG. 1 is a horizontal three-dimensional view of a delivery device.
FIG. 2 is a view similar to FIG. 1, showing the inside thereof.
FIG. 3 is a conceptual diagram of another embodiment of the delivery device according to the present invention.

Claims (1)

A therapeutic combination product used in the prevention and / or treatment of asthma, comprising:
(A) A drug containing a surfactant phospholipid (SAPL) composition in a finely divided form, where the surfactant phospholipid (SAPL) is on the surface in the vicinity of normal mammalian body temperature. Including an ingredient that enhances the diffusion of the drug in (b), and (b) an antiasthma drug,
The component (a) is selected from the group consisting of a first component containing one or more phosphatidylcholines and phosphatidylglycerols, phosphatidylethanolamines, phosphatidylserines, phosphatidylinositols, and chlorestyl palmitates A second component comprising one or more compounds,
At least said component (a) is arranged to be delivered to a patient in a dosage form that can be inhaled by at least one individual, the individual or each individual taking at least 25 mg of said component (a) Including
Wherein components (a) and said (b) is provided in the form of administration that together or separated in the form of a dosage,
Combination product wherein component (a) is that to form the adsorption layer of phospholipids spread over the surface of the lungs and airways.

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