Development of (Inhalable) Dry Powder Formulations of AS01(B)-Containing Vaccines Using Thin-Film Freeze-Drying
使用薄膜冷冻干燥技术开发含AS01(B)疫苗的（可吸入）干粉制剂
📄 英文摘要 English Abstract
AS01B is a liposomal formulation of two immunostimulants namely 3-O-desacyl-4́-monophosphoryl lipid A (MPL) and QS-21. The liposomal formulation of AS01B reduces the endotoxicity of MPL and the lytic activity of QS-21. The AS01B-adjuvanted Shingrix vaccine is marketed in a two-vial presentation, with the liquid AS01B liposomes in one vial and the antigen as a dry powder in another vial. In the present study, we tested the feasibility of applying thin-film freeze-drying (TFFD) to engineer dry powders of the AS01B liposomal adjuvant alone or vaccines containing AS01B as an adjuvant. Initially, we showed that after the AS01B liposomal adjuvant was subjected to TFFD using sucrose as a stabilizer at 4% w/v, the particle size distribution of AS01B liposomes reconstituted from the dry powder was identical to the liquid adjuvant before drying. We then showed using ovalbumin (OVA) as a model antigen adjuvanted with AS01B (AS01B/OVA) that subjecting the AS01B/OVA vaccine to TFFD and subsequent reconstitution did not negatively affect the AS01B liposome particle size, nor the immunogenicity of the vaccine. Importantly, the thin-film freeze-dried AS01B/OVA vaccine, unlike its liquid counterpart, was not sensitive to repeated freezing-and-thawing. The developed AS01B/OVA dry powder also showed the desirable aerosol properties (i.e., fine particle fraction of 66.3 ± 4.9% and mass median aerodynamic diameter of 2.4 ± 0.1 µm) for potential pulmonary administration. Finally, the feasibility of using TFFD to prepare dry powders of AS01B-adjuvanted vaccines was further confirmed using AS01B-adjuvanted Fluzone Quadrivalent and Shingrix, which contains AS01B. It is concluded that the TFFD technology can enable the formulation of AS01B-adjuvanted vaccines as freezing-insensitive, inhalable dry powders in a single-vial presentation. Keywords: Freeze-drying; Immunogenicity; Liposomes; Powder; Pulmonary; Single-vial; Vaccines. Copyright © 2022 Elsevier B.V. All rights reserved. PubMed Disclaimer MeSH terms Adjuvants, Immunologic Actions Search in PubMed Search in MeSH Add to Search
📄 中文摘要 Chinese Abstract
📋 英文结构化总结 English Structured Summary
摘要整理
**Header: Background** AS01B is a liposomal formulation of two immunostimulants namely 3-O-desacyl-4́-monophosphoryl lipid A (MPL) and QS-21. The liposomal formulation of AS01B reduces the endotoxicity of MPL and the lytic activity of QS-21. The AS01B-adjuvanted Shingrix vaccine is marketed in a two-vial presentation, with the liquid AS01B liposomes in one vial and the antigen as a dry powder in another vial. In the present study, we tested the feasibility of applying thin-film freeze-drying (TFFD) to engineer dry powders of the AS01B liposomal adjuvant alone or vaccines containing AS01B as an adjuvant.
**Header: Methods** Initially, we showed that after the AS01B liposomal adjuvant was subjected to TFFD using sucrose as a stabilizer at 4% w/v, the particle size distribution of AS01B liposomes reconstituted from the dry powder was identical to the liquid adjuvant before drying. We then showed using ovalbumin (OVA) as a model antigen adjuvanted with AS01B (AS01B/OVA) that subjecting the AS01B/OVA vaccine to TFFD and subsequent reconstitution did not negatively affect the AS01B liposome particle size, nor the immunogenicity of the vaccine. Finally, the feasibility of using TFFD to prepare dry powders of AS01B-adjuvanted vaccines was further confirmed using AS01B-adjuvanted Fluzone Quadrivalent and Shingrix, which contains AS01B.
**Header: Results** Initially, we showed that after the AS01B liposomal adjuvant was subjected to TFFD using sucrose as a stabilizer at 4% w/v, the particle size distribution of AS01B liposomes reconstituted from the dry powder was identical to the liquid adjuvant before drying. We then showed using ovalbumin (OVA) as a model antigen adjuvanted with AS01B (AS01B/OVA) that subjecting the AS01B/OVA vaccine to TFFD and subsequent reconstitution did not negatively affect the AS01B liposome particle size, nor the immunogenicity of the vaccine. Importantly, the thin-film freeze-dried AS01B/OVA vaccine, unlike its liquid counterpart, was not sensitive to repeated freezing-and-thawing. The developed AS01B/OVA dry powder also showed the desirable aerosol properties for potential pulmonary administration. Finally, the feasibility of using TFFD to prepare dry powders of AS01B-adjuvanted vaccines was further confirmed using AS01B-adjuvanted Fluzone Quadrivalent and Shingrix, which contains AS01B.
**Header: Data Summary** The developed AS01B/OVA dry powder also showed the desirable aerosol properties (i.e., fine particle fraction of 66.3 ± 4.9% and mass median aerodynamic diameter of 2.4 ± 0.1 µm) for potential pulmonary administration. Sucrose was used as a stabilizer at 4% w/v.
**Header: Conclusions** It is concluded that the TFFD technology can enable the formulation of AS01B-adjuvanted vaccines as freezing-insensitive, inhalable dry powders in a single-vial presentation.
**Header: Practical Significance** The developed AS01B/OVA dry powder also showed the desirable aerosol properties (i.e., fine particle fraction of 66.3 ± 4.9% and mass median aerodynamic diameter of 2.4 ± 0.1 µm) for potential pulmonary administration. The TFFD technology can enable the formulation of AS01B-adjuvanted vaccines as freezing-insensitive, inhalable dry powders in a single-vial presentation.
📋 中文结构化总结 Chinese Structured Summary
背景:
AS01B是一种包含两种免疫刺激剂——3-O-脱酰基-4'-单磷酰脂质A（MPL）和QS-21——的脂质体制剂。AS01B的脂质体制剂可降低MPL的内毒素活性和QS-21的溶血活性。含AS01B佐剂的Shingrix疫苗以双瓶形式上市销售，其中一瓶为液态AS01B脂质体，另一瓶为抗原冻干粉。在本研究中，我们测试了应用薄膜冷冻干燥（TFFD）技术制备AS01B脂质体佐剂单独或含AS01B佐剂疫苗干粉的可行性。
方法:
首先，我们证明了在以4% w/v蔗糖作为稳定剂的条件下对AS01B脂质体佐剂进行TFFD处理后，从干粉复溶的AS01B脂质体的粒径分布与干燥前的液态佐剂完全一致。随后，我们以卵清蛋白（OVA）作为模型抗原，与AS01B配伍（AS01B/OVA），证明对AS01B/OVA疫苗进行TFFD处理及后续复溶不会对AS01B脂质体粒径产生负面影响，也不会影响疫苗的免疫原性。最后，我们进一步利用含AS01B佐剂的四价流感疫苗Fluzone Quadrivalent和含AS01B的Shingrix疫苗，证实了使用TFFD制备含AS01B佐剂疫苗干粉的可行性。
结果:
首先，我们证明了在以4% w/v蔗糖作为稳定剂的条件下对AS01B脂质体佐剂进行TFFD处理后，从干粉复溶的AS01B脂质体的粒径分布与干燥前的液态佐剂完全一致。随后，我们以卵清蛋白（OVA）作为模型抗原，与AS01B配伍（AS01B/OVA），证明对AS01B/OVA疫苗进行TFFD处理及后续复溶不会对AS01B脂质体粒径产生负面影响，也不会影响疫苗的免疫原性。重要的是，薄膜冷冻干燥的AS01B/OVA疫苗与其液态对应物不同，对反复冻融不敏感。所开发的AS01B/OVA干粉还表现出良好的雾化特性，适合潜在的肺部给药。最后，我们进一步利用含AS01B佐剂的四价流感疫苗Fluzone Quadrivalent和含AS01B的Shingrix疫苗，证实了使用TFFD制备含AS01B佐剂疫苗干粉的可行性。
数据摘要:
所开发的AS01B/OVA干粉还表现出良好的雾化特性（即细颗粒分数为66.3 ± 4.9%，质量中位空气动力学直径为2.4 ± 0.1 µm），适合潜在的肺部给药。蔗糖以4% w/v的浓度用作稳定剂。
结论:
综上所述，TFFD技术能够将含AS01B佐剂的疫苗制备为对冷冻不敏感、可吸入的干粉制剂，并以单瓶形式呈现。
实际意义:
所开发的AS01B/OVA干粉还表现出良好的雾化特性（即细颗粒分数为66.3 ± 4.9%，质量中位空气动力学直径为2.4 ± 0.1 µm），适合潜在的肺部给药。TFFD技术能够将含AS01B佐剂的疫苗制备为对冷冻不敏感、可吸入的干粉制剂，并以单瓶形式呈现。