Manufacturing Stable Bacteriophage Powders by Including Buffer System in Formulations and Using Thin Film Freeze-drying Technology.
Manufacturing Stable Bacteriophage Powders by Including Buffer System in Formulations and Using Thin Film Freeze-drying Technology.
复制标题
通过在配方中加入缓冲系统并使用薄膜冷冻干燥技术制造稳定的噬菌体粉末。
DOI：
10.1007/s11095-021-03111-y
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发表时间：
2021
影响因子：
3.7
通讯作者：
Williams3rd,RobertO
中科院分区：
文献类型：
--
作者：
Zhang,Yajie;Soto,Melissa;Ghosh,Debadyuti;Williams3rd,RobertO
目的近年来，由于多重耐药“超级细菌”的流行，噬菌体治疗重新受到关注。为了开发治疗性噬菌体临床上可用的药物产品，已实施的策略固化噬菌体制剂多样化的剂型，并克服液体噬菌体formulation.MethodIn我们的工作中的存储条件的限制，我们假设和测试，一种先进的技术，薄膜冷冻干燥（TFFD），可用于生产噬菌体含有干粉，而不会显着失去噬菌体的活力。在这里，我们选择T7噬菌体作为我们的模型噬菌体在一个初步的筛选study.ResultsWe发现，一个二元辅料基质的蔗糖和亮氨酸的比例为90：10或75：25重量，保护噬菌体从应力期间遇到的TFFD过程。此外，我们证实，在制剂中掺入缓冲系统显著改善了在固化过程中的初始冷冻步骤和随后的升华步骤期间噬菌体的存活。在含有SM缓冲液（Tris/NaCl/MgSO 4）的制剂中噬菌体的滴度损失低至0.19 log噬菌斑形成单位，这表明在TFFD过程后噬菌体功能得到良好保留。缓冲液的存在显著降低了几何颗粒尺寸，如通过使用激光衍射的干分散法所测定的，这表明TFFD噬菌体粉末制剂容易剪切成较小的粉末聚集体，这是促进各种局部药物递送途径的理想性质，包括通过干粉吸入器的肺部递送、重构后的雾化和鼻内或伤口治疗，结论缓冲系统的引入可以稳定噬菌体的脱水过程，TFFD作为一种新的颗粒工程方法，可以成功地制备具有生物活性和潜在吸入治疗的含噬菌体的粉末。
PurposeBacteriophage (phage) therapy has re-gained attention lately given the ever-increasing prevalence of multi-drug resistance ‘super-bugs’. To develop therapeutic phage into clinically usable drug products, the strategy of solidifying phage formulations has been implemented to diversify the dosage forms and to overcome the storage condition limitations for liquid phage formulations.MethodIn our work, we hypothesize and tested that an advanced technology, thin film freeze-drying (TFFD), can be used to produce phage containing dry powders without significantly losing phage viability. Here we selected T7 phage as our model phage in a preliminary screening study.ResultsWe found that a binary excipient matrix of sucrose and leucine at ratios of 90:10 or 75:25 by weight, protected phage from the stresses encountered during the TFFD process. In addition, we confirmed that incorporating a buffer system in the formulation significantly improved the survival of phage during the initial freezing step and subsequent sublimation step in the solidifying processes. The titer loss of phage in SM buffer (Tris/NaCl/MgSO4) containing formulation was as low as 0.19 log plaque forming units, which indicated that phage function was well preserved after the TFFD process. The presence of buffers markedly reduced the geometric particle sizes as determined by a dry dispersion method using laser diffraction, which indicated that the TFFD phage powder formulations were easily sheared into smaller powder aggregates, an ideal property for facilitating a variety of topical drug delivery routes including pulmonary delivery through dry powder inhalers, nebulization after reconstitution, and intranasal or wound therapy, etc.ConclusionFrom these findings, we show that introducing buffer system can stabilize phage during dehydration processes, and TFFD, as a novel particle engineering method, can successfully produce phage containing powders that possess the desired properties for bioactivity and potentially for inhalation therapy.