A Dry Powder Formulation of Liposome-Encapsulated Recombinant Secretory Leukocyte Protease Inhibitor (rSLPI) for Inhalation: Preparation and Characterisation
A Dry Powder Formulation of Liposome-Encapsulated Recombinant Secretory Leukocyte Protease Inhibitor (rSLPI) for Inhalation: Preparation and Characterisation
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DOI：
10.1208/s12249-010-9500-2
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发表时间：
2010-09-01
影响因子：
3.3
通讯作者：
Cryan, Sally-Ann
中科院分区：
文献类型：
--
作者：
Gibbons, Aileen;McElvaney, Noel G.;Cryan, Sally-Ann
吸入重组分泌性白细胞蛋白酶抑制剂（rSLPI）已显示出治疗炎症性肺部疾病的潜力。迄今为止，组织蛋白酶L（Cat L）对rSLPI的快速灭活和从肺中的快速清除具有有限的临床功效。我们以前的研究表明，将rSLPI包封在1，2-二油酰基-sn-甘油-3-[磷酸-L-丝氨酸]/胆固醇（DOPS/Chol）脂质体中可保护rSLPI免受体外Cat L失活。发现液体DOPS-rSLPI制剂在长期储存和雾化后不稳定。因此，本研究的目的是开发一种用于制备吸入用干粉形式的DOPS-rSLPI脂质体的生产方法。将DOPS-rSLPI干粉冻干，随后用新型微粉化助剂微粉化。表征了制剂和加工对rSLPI稳定性、活性和粉末内含量均匀性的影响。使用D-甘露醇作为微粉化助剂，制备可吸入尺寸范围（< 5 μ m）的干粉颗粒。通过优化工艺参数，微粉化后回收了高达54%的rSLPI，其中抗中性粒细胞弹性蛋白酶活性没有显著损失，也没有可检测到的蛋白质降解证据。使用干粉吸入器实现雾化，并在级联撞击式采样器中收集后评价质量中值空气动力学直径（MMAD）。DOPS-rSLPI干粉的雾化产生38%的发射剂量，MMAD为2.44 μ m。当在雾化后用Cat L激发时，与水性DOPS-rSLPI脂质体分散体相比，DOPS-rSLPI干粉在保持针对Cat L诱导的rSLPI失活的保护功能方面显著更好，并且在储存下也更稳定。
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 mu m) 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 mu m 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.