Komárková Marie, Benešík Martin, Procházková Tereza, Vinco Adam, Laichmanová Monika, Smetanová Soňa, Jelínek Petr, Moša Marek, Botka Tibor, Šoóš Miroslav, Pantůček Roman
Department of Experimental Biology, Division of Genetics and Molecular Biology, Faculty of Science, Masaryk University, 611 37 Brno, Czech Republic.
MB Pharma s.r.o., 120 00 Prague, Czech Republic.
Eur J Pharm Biopharm. 2025 Sep;214:114781. doi: 10.1016/j.ejpb.2025.114781. Epub 2025 Jun 10.
Implementing bacteriophages into dosage forms is a significant step for the practical application of phage therapy. While designing a dosage form, bacteriophages as active ingredients may be exposed to excipients, guaranteeing microbial quality. However, only a few antimicrobial preservatives have been studied regarding their interaction with bacteriophages during long-term storage. Here, the stability of the staphylococcal Kayvirus and pseudomonal Pbunavirus with twelve commonly used preservatives was monitored for thirteen weeks to assess the risk of destabilisation of phage suspensions by excipients. The effectiveness of preservatives on the test bacteria, yeast and mould was determined using a microdilution method and the phage lytic activity by plaque enumeration. The antimicrobial activity of preservatives with bacteriophages was confirmed, except benzalkonium chloride and chlorhexidine digluconate, which showed precipitation and were classified as incompatible. A complete loss of phage potency in both tested phages occurred with diazolidinyl urea and in Kayvirus with benzalkonium chloride. For both phages, a slight decrease in titer, by one order of magnitude, was observed with m-cresol, sodium propionate, sodium benzoate, and phenylethyl alcohol. For Kayvirus, thimerosal, parabens, and mono propylene glycol and for Pbunavirus, phenoxyethanol also met the criteria. The decrease by two or more orders was determined for the remaining cases. This study helps select antimicrobial preservatives for optimizing dosage formulations with the therapeutically applicable bacteriophages.
将噬菌体应用于剂型是噬菌体疗法实际应用的重要一步。在设计剂型时，作为活性成分的噬菌体可能会接触到辅料，以保证微生物质量。然而，关于少数抗菌防腐剂在长期储存过程中与噬菌体的相互作用，目前研究较少。在此，监测了葡萄球菌Kayvirus和铜绿假单胞菌Pbunavirus与十二种常用防腐剂的稳定性，为期十三周，以评估辅料导致噬菌体悬浮液失稳的风险。采用微量稀释法测定防腐剂对受试细菌、酵母菌和霉菌的有效性，并通过噬菌斑计数法测定噬菌体的裂解活性。除苯扎氯铵和葡萄糖酸氯己定出现沉淀并被归类为不相容外，证实了防腐剂与噬菌体的抗菌活性。在两种受试噬菌体中，双咪唑烷基脲会导致噬菌体效力完全丧失，在Kayvirus中，苯扎氯铵也会导致噬菌体效力完全丧失。对于两种噬菌体，间甲酚、丙酸钠、苯甲酸钠和苯乙醇会使其滴度略有下降，下降一个数量级。对于Kayvirus，硫柳汞、对羟基苯甲酸酯和单丙二醇，以及对于Pbunavirus，苯氧乙醇也符合标准。其余情况下，噬菌体滴度下降两个或更多数量级。本研究有助于选择抗菌防腐剂，以优化含治疗用噬菌体的剂型配方。
