脂质体包封的重组分泌型白细胞蛋白酶抑制剂(rSLPI)干粉吸入制剂的制备与表征。
A dry powder formulation of liposome-encapsulated recombinant secretory leukocyte protease inhibitor (rSLPI) for inhalation: preparation and characterisation.
机构信息
School of Pharmacy, Royal College of Surgeons in Ireland, York House, York Street, Dublin 2, Ireland.
出版信息
AAPS PharmSciTech. 2010 Sep;11(3):1411-21. doi: 10.1208/s12249-010-9500-2. Epub 2010 Sep 15.
Inhaled recombinant secretory leukocyte protease inhibitor (rSLPI) has shown potential for the treatment of inflammatory lung conditions. Rapid inactivation of rSLPI by cathepsin L (Cat L) and rapid clearance from the lungs has limited clinical efficacy to date. Previous studies by us have shown that encapsulation of rSLPI within1,2-dioleoyl-sn-glycero-3-[phospho-L-serine]/cholesterol (DOPS/Chol) liposomes protects rSLPI against Cat L inactivation in vitro. Liquid DOPS-rSLPI preparations were found to be unstable upon long-term storage and nebulisation. The aim of this study was therefore to develop a method of manufacture for preparing DOPS-rSLPI liposomes as a dry powder for inhalation. DOPS-rSLPI dry powders were lyophilised and subsequently micronised with a novel micronisation aid. The effects of formulation and processing on rSLPI stability, activity, and uniformity of content within the powders were characterised. Using D-mannitol as the micronisation aid, dry powder particles in the inhalable size range (<5 μm) were prepared. By optimising process parameters, up to 54% of rSLPI was recovered after micronisation, of which there was no significant loss in anti-neutrophil elastase activity and no detectable evidence of protein degradation. Aerosolisation was achieved using a dry powder inhaler, and mass median aerodynamic diameter (MMAD) was evaluated after collection in a cascade impactor. Aerosolisation of the DOPS-rSLPI dry powder yielded 38% emitted dose, with 2.44 μm MMAD. When challenged with Cat L post-aerosolisation, DOPS-rSLPI dry powder was significantly better at retaining a protective function against Cat L-induced rSLPI inactivation compared to the aqueous DOPS-rSLPI liposome dispersion and was also more stable under storage.
吸入重组分泌型白细胞蛋白酶抑制剂(rSLPI)已显示出治疗炎症性肺病的潜力。迄今为止,rSLPI 被组织蛋白酶 L(Cat L)迅速失活和从肺部迅速清除,限制了其临床疗效。我们之前的研究表明,将 rSLPI 包裹在 1,2-二油酰基-sn-甘油-3-[磷酸-L-丝氨酸]/胆固醇(DOPS/Chol)脂质体中可防止 rSLPI 在体外被 Cat L 失活。长期储存和雾化时,液态 DOPS-rSLPI 制剂被发现不稳定。因此,本研究旨在开发一种制备 DOPS-rSLPI 脂质体作为干粉吸入剂的方法。DOPS-rSLPI 干粉经冷冻干燥后,用新型助磨剂进行微粉化。研究了制剂和加工对 rSLPI 稳定性、活性和粉末中含量均匀性的影响。使用 D-甘露醇作为助磨剂,可制备出可吸入粒径范围(<5μm)的干粉颗粒。通过优化工艺参数,可在微粉化后回收高达 54%的 rSLPI,其抗中性粒细胞弹性蛋白酶活性无明显损失,也没有检测到蛋白质降解的迹象。干粉吸入器可实现气溶胶化,在级联撞击器中收集后评估质量中值空气动力学直径(MMAD)。DOPS-rSLPI 干粉的气溶胶化产生 38%的发射剂量,2.44μm 的 MMAD。气溶胶化 DOPS-rSLPI 干粉后,与 DOPS-rSLPI 脂质体分散液相比,其对 Cat L 诱导的 rSLPI 失活具有更好的保护作用,并且在储存过程中也更加稳定。