Cowen Todd, Walmsley Simon, Karim Kal, Haser Resul, Caffrey Patrick, Piletska Elena, Rawlings Bernard, Piletsky Sergey A
School of Chemistry, University of Leicester, Leicester LE1 7RH, UK.
School of Biomolecular and Biomedical Science, University College Dublin, D04 V1W8 Dublin, Ireland.
Biomimetics (Basel). 2023 Jun 27;8(3):273. doi: 10.3390/biomimetics8030273.
Amphotericin B has been an essential drug in the fight against leishmaniasis and fungal pathogens for decades, and has more recently gained attention for the very limited microbial resistance displayed against it. However, its toxicity has restricted its use to only the most severe cases of disease, and attempts to reduce these ill effects via formulation have had only minor success. Genetic engineering has allowed the development of superior amphotericin analogues, notably 16-descarboxyl-16-methyl amphotericin B (MeAmB), which shows a ten-fold reduction in toxicity in addition to a slight improvement in therapeutic activity. However, MeAmB is difficult to extract from its bacterial source and purify. Presented here is an alternative method of MeAmB purification. A biomimetic polymer with a high affinity for MeAmB was designed via computational modelling and synthesised. Prepared as a separation column, the polymer was able to retain the target MeAmB whilst allowing the removal of cell debris from the bacterial extract. Starting with a simple bacterial extract, the relatively simple process allowed the purification of an MeAmB salt complex at approximately 70% MeAmB, and likely higher purification from further extraction. The mean MeAmB recovery between the pre-purification extract sample and the final product was 81%. This is the first successful demonstration of extraction or purification of any amphotericin molecule with any polymeric material. The biomimetic polymer was additionally reusable and simple to fabricate, giving this technique significant advantages over traditional methods of extraction and purification of valuable compounds.
几十年来，两性霉素B一直是对抗利什曼病和真菌病原体的重要药物，最近它因对其产生的微生物耐药性非常有限而受到关注。然而，其毒性限制了它仅用于最严重的疾病病例，并且通过制剂来减少这些不良影响的尝试仅取得了微小的成功。基因工程使得能够开发出更优良的两性霉素类似物，特别是16-去羧基-16-甲基两性霉素B（MeAmB），它除了治疗活性略有提高外，毒性降低了十倍。然而，MeAmB难以从其细菌来源中提取和纯化。本文介绍了一种纯化MeAmB的替代方法。通过计算建模设计并合成了一种对MeAmB具有高亲和力的仿生聚合物。将该聚合物制备成分离柱后，它能够保留目标MeAmB，同时允许从细菌提取物中去除细胞碎片。从简单的细菌提取物开始，这个相对简单的过程能够纯化出MeAmB盐复合物，其中MeAmB含量约为70%，进一步提取可能会得到更高的纯度。纯化前提取物样品与最终产物之间的MeAmB平均回收率为81%。这是首次成功展示用任何聚合材料提取或纯化任何两性霉素分子。这种仿生聚合物还可重复使用且易于制备，使得这项技术相对于传统的有价值化合物提取和纯化方法具有显著优势。
