Abstract

Capreomycin (CMN) is a second-line aminoglycoside antibiotic for treating multi-drug resistant tuberculosis. Produced as a complex broth in microbial bioreactors, the downstream purification of CMN traditionally requires multiple steps. Current work aims to purify CMN as mixed isomers in a single step by preparative macroporous cation-exchange chromatography in >98% purity in >90% yield. Selection of the adsorbent best suited for CMN purification was guided by modeling the molecular interactions, followed by wet-lab static and dynamic adsorption studies that validated the simulation results. Among the evaluated adsorbents (both weak and strong cation resins), the weak cationic Macro-Prep CM showed a favorable combination of selective binding and elution of CMN. Solute loading and elution conditions were further optimized to produce the desired quality of CMN. This single-step method was integrated with a continuous simulated bed reverse-phase desalting chromatography to produce 99% pure salt-free CMN. The proposed method provided productivity of about 2 g product/L resin/h of CMN on Macro-Prep CM; and 0.7 g product/L adsorbent/h on SMB. The study provides for a robust and scalable process for the industrial-scale production of purified capreomycin.

摘要

卷曲霉素 (CMN) 是一种用于治疗耐多药结核病的二线氨基糖苷类抗生素。作为微生物生物反应器中的复杂肉汤生产，CMN 的下游纯化传统上需要多个步骤。目前的工作旨在通过制备型大孔阳离子交换色谱法以 >98% 的纯度和 >90% 的产率在一步中纯化混合异构体形式的 CMN。通过对分子相互作用建模，然后进行湿实验室静态和动态吸附研究来验证模拟结果，从而指导选择最适合 CMN 纯化的吸附剂。在评估的吸附剂（弱和强阳离子树脂）中，弱阳离子 Macro-Prep CM 表现出对 CMN 的选择性结合和洗脱的有利组合。进一步优化溶质加载和洗脱条件以产生所需质量的 CMN。这种单步方法与连续模拟床反相脱盐色谱相结合，可生产 99% 纯度的无盐 CMN。所提出的方法在 Macro-Prep CM 上提供了约 2 g 产品/L 树脂/小时 CMN 的生产率；和 0.7 克产品/L 吸附剂/小时，在 SMB 上。该研究为纯化卷曲霉素的工业规模生产提供了稳健且可扩展的过程。